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Survivin is a member of the inhibitor of apoptosis protein (IAP) family. Survivin has been reported to be expressed in many cancers, but not in differentiated normal tissue. Recent studies revealed that survivin correlated with the chemo-resitance of cancer cells. In the present study, the changes in expression levels of survivin messenger RNA (mRNA) and survivin protein in a gastric cancer cell line (MKN-45) during cisplatin (CDDP) treatment were analyzed and compared with the occurrence of apoptotic cell death. Cell growth was inhibited even with a low dose CDDP (0.1 or 1 microg/ml) 1 hr treatment. However, the percentage of apoptotic cells did not change after 48 hr incubation with low dose CDDP. Only with high dose CDDP (10 microg/ml), did the percentage of apoptotic cells explosively increase between 12 and 24 hr treatment. Relative expression levels of survivin mRNA and survivin protein increased after CDDP treatment. The cell expression rates of survivin mRNA after 48 hr treatment with 0.1 and 1 microg/ml of CDDP were 2 to 6 fold higher than that of the survivin mRNA of untreated cells. Also, the relative cell expression level of survivin protein after 24 hr treatment with 0.1 or 1 microg/ml of CDDP was 3 to 6.5 fold higher than that of the survivin protein of untreated cells. These results indicate that survivin expression may correlate with the chemo-resistance of malignant cells.

Five lines (MKN-28, MKN-45, MKN-74, KATO-III, and HSC-39) of human gastric carcinoma cells were treated with 1 microgram/ml cisplatin for 12, 24, 36, 48, and 72 h. Apoptotic indices (percentages of apoptotic cells) were analyzed at each time point. After incubation with cisplatin, apoptotic cells were detected more frequently among MKN-45 and MKN-74 cells with a wild-type gene for p53 and without expression of the bcl-2 protein than among HSC-39, MKN-28, and KATO-III cells with a mutation or complete deletion of the gene for p53 and with overexpression of the bcl-2 protein. The levels of p53 protein increased after treatment with cisplatin in MKN-74 cells. However, levels of bcl-2 protein were reduced in KATO-III after treatment with cisplatin. Thus, p53 status and the expression of bcl-2 by tumor cells might be good indicators of sensitivity to chemotherapy for patients with gastric cancer.

Hepatocellular carcinoma (HCC) is one of the most common human malignancies in the world and it is especially prevalent in Asia and Africa (1). Some tumors are suitable for resection, however, there is a very high rate of recurrence (2). Infection with hepatitis B or C virus, alcoholic cirrhosis, and ingestion of aflatoxin B1-contaminated food are important risk factors for HCC (3,4), but the exact molecular mechanism of tumorigenesis is still unclear.

The cadherin-catenin complex has been recognized as an important factor associated with tumor metastasis. However, the clinical significance of the expression of adhesion molecules in lymph nodes with metastasis remains unclear. The aim of this study was to investigate the clinical significance of the re-expression of the cadherin-catenin complex in metastatic lymph nodes in patients with advanced gastric cancer. Immunohistochemical expression of E-cadherin, alpha- and beta-catenin were analyzed in 96 primary gastric cancers with serosal invasion and in 79 lymph nodes with metastasis. The expression levels of these adhesion molecules in primary tumors and lymph nodes with metastasis were compared. Ninety-four out of 96 primary tumors (98%) showed reduced expression of adhesion molecules. Out of 79 cases with lymph node metastasis, increased expression of one or more adhesion molecules in metastatic foci as compared with primary tumors was detected in 52 cases (66%). Re-expression of adhesion molecules in metastatic lymph nodes was detected in a more advanced stage. The overall 5-year survival rate of the 52 patients who had lymph nodes with metastasis with re-expression of adhesion molecules (8%) was significantly poorer than that of the 27 who had lymph nodes with metastasis without re-expression of adhesion molecules (33%, P = 0.0012). The re-expression of the cadherin-catenin complex in lymph nodes with metastasis may play an important role in the growth of cancer cells in metastatic foci. A comparison of the expression patterns of adhesion molecules between the primary tumor and metastatic lymph nodes may provide new prognostic information for patients with advanced gastric cancer.

Although surgical resectability is an important prognostic factor, recurrences are commonly noted in advanced colorectal cancer patients, even after apparently curative surgery. Since such recurrences cannot be cured, better adjuvant chemotherapies are urgently required.We studied the effect of post-operative chemotherapy using oral administration of 1-hexylcarbamoyl-5-fluorouracil (HCFU) with 5-fluorouracil (5-FU) infusion for curatively-resected Stage IIIa and IIIb colorectal cancers. This study was prospectively randomized and controlled and 314 (97.8%) out of 321 patients were determined to be candidates for statistical assessment. Group A and Group B received 5-FU intravenous injection at, respectively, 333 mg/m2 and 1000 mg/m2 body surface area/24 hours continuously for 72 hours beginning on post-operative day 0 and day 6, with oral HCFU 300 mg daily for 52 weeks beginning 2 weeks after surgery.There were no differences in overall 5-year survival or disease-free survival between Group A and Group B. A retrospective subset analysis. however, suggested that the protocol of Group B tended to yield better 5-year survival (68.3%) for rectal cancer than that of Group A (58.8%).Inductive therapy with high-dose 5-FU in combination with oral HCFU appears to be beneficial as adjuvant chemotherapy for advanced rectal cancer with lymph node metastasis.

Survivin is a member of the inhibitor of apoptosis protein (IAP) family. Survivin has been reported to be expressed in many cancers, but not in differentiated normal tissue. Recent studies revealed that survivin correlated with the chemo-resitance of cancer cells. In the present study, the changes in expression levels of survivin messenger RNA (mRNA) and survivin protein in a gastric cancer cell line (MKN-45) during cisplatin (CDDP) treatment were analyzed and compared with the occurrence of apoptotic cell death. Cell growth was inhibited even with a low dose CDDP (0.1 or 1 microg/ml) 1 hr treatment. However, the percentage of apoptotic cells did not change after 48 hr incubation with low dose CDDP. Only with high dose CDDP (10 microg/ml), did the percentage of apoptotic cells explosively increase between 12 and 24 hr treatment. Relative expression levels of survivin mRNA and survivin protein increased after CDDP treatment. The cell expression rates of survivin mRNA after 48 hr treatment with 0.1 and 1 microg/ml of CDDP were 2 to 6 fold higher than that of the survivin mRNA of untreated cells. Also, the relative cell expression level of survivin protein after 24 hr treatment with 0.1 or 1 microg/ml of CDDP was 3 to 6.5 fold higher than that of the survivin protein of untreated cells. These results indicate that survivin expression may correlate with the chemo-resistance of malignant cells.

We studied whether the immunohistochemical status of dihydropyrimidine dehydrogenase (DPD) and p53 can be used to predict the sensitivity to chemoradiotherapy (CRT) in patients with esophageal cancer. In 19 patients who did not undergo preoperative CRT, the immunoreactivity of DPD and p53 in biopsied specimens correlated well with those in surgically resected specimens (DPD: 100%, p53: 73%). Fifteen patients were treated with 5-FU (250-300 mg/body/day: day 1-5, 8-12), low-dose cisplatin (10 mg/body/day: day 1, 8) and radiotherapy (30-40 Gy). The response rate (CR + PR) for CRT in these patients was 40%. All tumors that showed CR or PR demonstrated low expression of DPD. However, all tumors with high DPD expression showed MR or NC. However, the expression of p53 did not correlate with the response rate for CRT. Therefore, the effect of CRT for esophageal cancer may be predicted by immunohistochemical examination of DPD in biopsied tumor specimens.

In hepatocellular carcinoma (HCC), new tumors develop in the residual liver within a few years after hepatectomy. However, the biological risk factors of multifocal occurrence of cancers remains unclear. In this study, the thymidine phosphorylase (TP) activity, which is known as an angiogenic factor, of cancerous and non-cancerous liver tissues in HCC was analyzed to determine its suitability as a biological marker of the multifocal occurrence of HCCs.Fresh tissues (tumor: HCC and adjacent liver tissue: N-HCC) from 63 patients with HCC and normal liver tissues (NL) from 6 patients without HCC were obtained. The TP activities of the tissues were analyzed by an enzyme-linked immunosorbent assay (ELISA).The mean TP activity of 63 HCCs (136 U/mg protein) was higher than that of 63 N-HCCs (81 U/mg protein) and that of 6 NLs (47 U/mg protein, p0.001). Multifocal occurrence of HCCs were detected in 17 patients. In these 17 patients, the mean TP activity of HCCs (145 U/mg protein) was not different from that of HCCs from the remaining 46 patients (133 U/mg protein, p = 0.272), however the mean TP activity of N-HCCs (110 U/mg protein) was significantly higher than that of N-HCCs from the remaining 46 patients (71 U/mg protein, p = 0.038). Moreover, only a high TP activity of N-HCCs was detected as a significant risk factor of multifocal occurrence of HCCs.Patients who have tumors with high TP activity in the non-cancerous livers may have a risk of multifocal occurrence of HCCs in the residual liver.

Micrometastases are considered to be a cause of recurrence after curative surgery for gastric cancer. It is important to clarify the clinicopathologic characteristics of micrometastases in the lymph nodes and peritoneal cavity to determine the treatment options in gastric cancer. Two consecutive sections of lymph nodes from patients with various cancers were examined by simultaneous staining with ordinary hematoxylin and eosin (HE) and immunostaining with anti-cytokeratin antibody, respectively. Micrometastases in the lymph nodes were found in 18% of mucosal cancer, 25% of submucosal cancer, and 65% of T3 (serosal) cancers pecimens, with cancer-free nodes examined by HE staining. A reduced 5-year survival rate was demonstrated in patients with nodal micrometastases among those with submucosal cancer and those with T3 cancer and cancer-free nodes examined by HE staining. Molecular biological detection (MBD) of micrometastasis in lavage cytology specimens was performed by RT-PCR of carcinoembryonic antigen mRNA or telomerase activity assay. MBD protocols revealed micrometastases in cases with negative cytology results. Survival analysis demonstrated peritoneal recurrences in MBD-positive cases, whereas there was no recurrence in MBD-negative cases. Peritoneal micrometastases detected by MBD protocols appear to be a significant risk factor for recurrence. Therefore indications for lymph node dissection and postoperative chemotherapy should be determined based on the findings of micrometastases in gastric cancer.

Surgical technique and postoperative care for gastric cancer have significantly improved in recent years. However, whether postoperative morbidity or mortality rates after gastrectomy for gastric cancer were reduced or not in recent years was unclear. In this study, we analyzed the chronological changes of postoperative morbidity and mortality rates, and we analyzed risk factors for postoperative morbidity and mortality in patients undergoing gastrectomy for carcinomas of the stomach.A total of 887 patients with gastric cancer were gastrectomized in our hospital between January 1985 and December 1996. The patients were divided into three groups on the basis of chronology. The first group included patients treated over the period 1985 to 1988 (n = 324); the second group, 1989 to 1992 (n = 300); and the third group, 1993 to 1996 (n = 263). Postoperative morbidity rates and mortality rates were compared among the three groups. Also, significant risk factors affecting postoperative morbidity and in-hospital mortality were analyzed by the multiple logistic regression analysis.Postoperative complications were detected in 95 patients (10.7%) and in-hospital mortality rate was 2.4% (21/887). Postoperative morbidity rates were 10.5%, 11%, and 10.6% in the first, second, and third groups, respectively and postoperative mortality rates were 2.5%, 2%, and 2.7%, respectively. These postoperative morbidity and mortality rates were not different between the groups (P = 0.979 and P = 0.866). The most common postoperative complication was anastomotic leakage (56/95, 58.9%). Significant risk factors affecting in-hospital mortality were Stage IV (P = 0.006) and noncurative gastric resection (P = 0.004). However, the extent of lymph node dissection, combined resection, or the existence of preoperative complications were not significant risk factors of in-hospital mortality by multiple logistic regression analysis.These results indicate that patients with far-advanced gastric cancer might have a high risk of postoperative mortality. In noncurative operations for patients with advanced gastric cancer, unnecessary lymph node dissection or combined resection should be avoided.

The relationship between prognostic factors and survival time after noncurative gastric resection in patients with advanced gastric cancer was examined by a retrospective review of data on 364 patients.There were 168 patients without metastasis to the liver or peritoneum (group A), 127 with peritoneal metastasis and no liver metastasis (group B), 50 with liver metastasis and no peritoneal metastasis (group C) and 19 with synchronous liver and peritoneal metastases (group D). Patients were primarily treated with the following 3 drugs: the fluorinated pyrimidines, cisplatin, and mitomycin C.Patients in group D had a very poor prognosis as compared with the other groups. Multivariate analysis using the Cox's proportional hazard model adjusted for sex, age, and other covariants indicated that lymph node metastasis, lymph node dissection, and fluorinated pyrimidines for group A, cisplatin for group B, and lymph node dissection for group C were independent prognostic factors. An analysis of patients excluding cases who died within 30 days after surgery revealed that lymph node dissection for group A, lymph node dissection and cisplatin for group B, and lymph node dissection for group C were independent prognostic factors.Treatment protocol specific for the residual disease may improve the survival of patients with advanced gastric cancer treated by noncurative resection.

The aim of this study was to investigate whether angiogenic factors influence the occurrence of spontaneous apoptosis in advanced gastric cancer. The apoptotic indices (AIs) of 97 tumors from 97 patients with advanced gastric cancer (pT3, pN0, pM0, Stage II) were analyzed by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling (TUNEL) method. Intratumoral microvessel densities (IMVDs) of tumors stained with anti-CD34 monoclonal antibody were quantified under x 200 magnification using computer-assisted image analysis. The expressions of angiogenic factors, such as vascular endothelial growth factor (VEGF), thymidine phosphorylase (dThdPase), transforming growth factor-alpha (TGF-alpha), and p53 were analyzed immunohistochemically and compared with IMVDs and AIs. The mean IMVD of the 97 tumors was 365/mm2 (range 147-990/mm2). The mean AI of tumors was 2.1% (range 0-11.3%). A significant inverse correlation between the AIs and the IMVDs was shown (p = -0.278, P = 0.0064). The mean IMVDs of tumors with high expressions of dThdPase, TGF-alpha, or p53 were significantly higher than those of tumors with low expressions of these factors. The mean AI of tumors with high expressions of dThdPase was significantly lower than that of tumors with low expressions of dThdPase (P = 0.023). However, no significant correlations were detected between AIs and the expression levels of VEGF, TGF-alpha, or p53. In gastric cancer, dThdPase may play an important role in tumor progression by increasing microvessels and by suppressing apoptosis of cancer cells.

The accumulation of mutated p53 in tumor cells results in the presence in the circulation of mutant p53 (p53m) and the production of p53-specific antibodies (p53Ab). We examined the relationships among these phenomena and analyzed their clinical implications in 62 patients with gastric cancer at various stages. Expression of p53 in tumors was studied by an immunohistochemical method and circulating p53m and p53Ab were quantitated with commercially available enzyme-linked immunosorbent assay kits. The detectable expression of p53 in tumors and circulating p53Ab was recognized in 28 (45.2%) and 7 (11.3%) of the 62 patients, respectively. The number of patients with higher levels of circulating p53m increased with the progression of the depth of cancer invasion. Patients with any positive findings for the three p53-related parameters had a poorer prognosis, and the difference was statistically significant in patients with p53Ab. When survival was analyzed in terms of the combination of the three p53-related parameters (detectable expression of p53 in tumor cells, high levels of p53m and p53Ab in the circulation), a significantly poorer prognosis was associated with an increase in the number of positive parameters. Analysis of p53 in tumor cells, together with analysis of circulating p53m and p53Ab, could improve the accuracy of prognosis in patients with gastric cancer.

Familial adenomatous polyposis (FAP) is a dominantly inherited disorder and it is difficult to know when polyps change to carcinoma. In this study, we retrospectively evaluated whether telomerase activity and p53 gene mutations are useful parameters for detecting malignant changes in polyps in patients with FAP.Twenty-one tissue samples (carcinoma: 5, large polyp: 8, and small polyp: 8) were obtained from 8 patients with FAP. Colorectal carcinomas were detected in 5 patients. As a control, 68 colorectal carcinomas and 11 adenomatous polyps with synchronous colorectal carcinoma from 68 patients and 8 polyps from 8 patients without colorectal carcinoma were obtained. The telomeric repeat amplification protocol (TRAP) assay was used for the detection of telomerase activity. In 21 samples from FAP patients, mutations of exon 5 to 8 in the p53 gene were analyzed by the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method.In 5 patients with FAP who had carcinomas in their colon and rectum, all the samples (carcinomas, large polyps and small polyps) showed telomerase activity. In contrast, in the 3 patients with FAP who had no carcinomas, telomerase activity was not detected in either the large or small polyps. Telomerase activity was detected in 51 out of 68 control colorectal carcinomas (75%) and in 6 out of 19 adenomas (32%). The p53 gene mutation was detected in only 2 carcinomas from 2 patients with FAP and was not detected in any large and small polyps.These findings suggest that telomerase is activated before the occurrence of the p53 gene mutation. In FAP, genetic changes associated with the development of colonic adenoma into carcinoma might activate telomerase and such genetic changes might occur in whole polyps of the colon and rectum.

Although many studies have attempted to clarify the prognostic indicators for gastric carcinoma, there have been few studies regarding the factors that correlate with the survival period of patients with postoperative recurrence.Among 504 advanced gastric adenocarcinoma patients who had undergone curative gastrectomy, 188 patients who had died of recurrence were used in this study.Univariate analysis indicated that age, the presence of lymph node metastasis and blood vessel invasion, the number of positive lymph nodes, and gastrectomy significantly correlated with the survival period. Multivariate analysis indicated that the length of the survival period was independently influenced by the number of positive lymph nodes and blood vessel invasion. The survival time of patients with less than 3 positive lymph nodes and no accompanying blood vessel invasion was significantly longer than that of other patients.The number of positive lymph nodes and the presence of blood vessel invasion are the most important factors predicting the survival period of patients with postoperative recurrence after curative resection for advanced gastric carcinoma.

Overexpression of transforming growth factor beta s (TGF-beta s), multifunctional polypeptides, have been reported in the tissue of various types of carcinomas, including gastric carcinoma. To our knowledge, serum levels of TGF-beta 1 have not previously been examined in patients with gastric carcinoma as a possible indicator of prognosis.Preoperative serum levels of TGF-beta 1 were measured in 111 patients with gastric carcinoma by an enzyme-linked immunosorbent assay.An elevated level of TGF-beta 1 was significantly correlated with lymph node metastasis and poor prognosis. Moreover, the preoperative serum levels of TGF-beta 1 in patients with peritoneal recurrence were significantly higher than those in patients both with lymph node recurrence and without recurrence.The serum level of TGF-beta 1 might be useful for predicting recurrence patterns and prognosis in patients with gastric carcinoma.

Synchronous and metachronous hematogenic metastases are frequently detected in patients with colorectal carcinoma. Once these metastases have developed, the prognoses of patients are poor. Previously, we reported that enlargement of cancer nuclei significantly correlated with metastatic potential of gastric cancer and hepatocellular carcinoma. Moreover, recently it has been reported that reduced expression of E-cadherin is associated with tumor metastasis. To evaluate the correlation between nuclear area (NA) of cancer cells and expression of E-cadherin, and to elucidate whether these factors correlate with clinical outcome in patients with colorectal carcinoma, 105 consecutive patients were investigated.In each case, the NAs of 600 cancer nuclei were analyzed by means of a computer-assisted image analysis system and E-cadherin expression was detected immunohistochemically by an anti-E-cadherin monoclonal antibody. The expression levels of E-cadherin were divided into three groups according to the percentages of E-cadherin-positive cells (level 0: positive cellsor = 50%, level 1: 50%positive cellsor = 80%, level 2: positive cells80%).The mean NA of cancer cells in 105 tumors was 57 microm2. The NAs of cancer cells enlarged in proportion to the decrease of E-cadherin expression levels (level 0, n = 48, 62 microm2; level 1, n = 35, 57 microm2, level 2, n = 22, 46 microm2, P = 0.002). The 10-year survival rates decreased in proportion to the reduced E-cadherin expression levels (80% in level 2, 64% in level 1, and 42% in level 0, P = 0.004). Moreover, the 10-year survival rate of 54 patients with large NA tumors (or = 54 microm2, 36%) was significantly poorer than that of 51 patients with small NA tumors (54 microm2, 80%, P0.001). The NA of cancer cells was recognized as an important predictor for prognosis and hematogenic metastasis. Although reduced E-cadherin expression was not recognized as the risk factor for hematogenic metastasis, 80% of patients who developed hematogenic metastasis had tumors with both enlarged cancer nuclei and reduced E-cadherin expression.Detection of NA of cancer cells and E-cadherin expression in patients with colorectal carcinoma may reveal important information for hematogenic metastasis.

Expression of thymidylate synthase (TS) has been studied as a prognostic factor and mechanism of drug resistance in gastric cancers. The relationship between TS expression in surgically resected specimens and clinicopathological factors was examined in 216 gastric cancer patients. Immunohistochemical demonstration of the protein was achieved using an anti-TS polyclonal antibody. Positive TS staining was observed in 50 patients (23.1%). Lymph node metastasis was more frequent in patients with TS-positive tumors than in those with TS-negative tumors (P0.01). Patients were followed for more than 5 years and survival was examined. In 163 patients who received fluorouracil (FU)-based chemotherapy, the overall 5-year survival rate was 41.8% for patients with TS-positive tumors and 57.0% for patients with TS-negative tumors (P0.01). In the 53 patients who did not receive chemotherapy, these figures were 25.6% and 79.5%, respectively (P0.05). In patients with T3 gastric cancer who were treated with curative gastrectomy, however, FU-based chemotherapy did not affect survival of either patients with TS-positive tumors or with TS-negative tumors. Multivariate analysis also revealed TS expression to be a significant variable for predicting postoperative survival (P0.05). These results indicate that TS expression can be used as an independent prognostic factor for patients with gastric cancer. However, TS expression is not a major predictor of the efficacy of FU-based chemotherapy.

We examined whether overexpression of p53 can be used as a new genetic marker to predict the presence of lymph node metastases of early invasive colorectal cancer. Forty-nine patients with primary colorectal adenocarcinomas invading to the submucosa (sm-CRC) were analyzed and 7 patients were found to have lymph node metastases. Immunostaining was used to detect the p53 overexpression; 43% of sm-CRC stained positive for p53 and all the cancer cells metastasized to lymph nodes were p53 positive. Both lymph node involvement and tumor budding were significantly more frequent in p53 positive than p53 negative tumors (p0.05, respectively), and multivariate analysis showed that p53 overexpression constituted a higher relative lisk for lymph node metastases of sm-CRC than either histologic type, level of sm invasion, macroscopic type, tumor budding or vascular invasion, although the difference was not significant (p = 0.086). We concluded that p53 overexpression is a useful biological marker of lymph node metastases of sm-CRC, and that p53 negative status may be an indicator for limited surgery, such as local excision of sm-CRC.

The pharmacokinetics of intraperitoneal (i.p.) and intrapleural (i.pl.) hypotonic cisplatin (CDDP) were compared under the same experimental conditions. The same dose of CDDP was administered in hypotonic (62 mOsm/L) and isotonic (308 mOsm/L) solutions to the peritoneal and pleural cavities of Ehrlich carcinoma cell bearing mice. The intracellular amount of platinum increased for more than 60 minutes after an i.pl. injection of the hypotonic solution of CDDP, whereas it increased for up to 30 minutes after an i.p. injection. Although hypotonic conditions augmented the amount of platinum taken-up by Ehrlich cells, the amount was significantly greater in the pleural cavity than in the peritoneal cavity. In Donryu rats, the levels of platinum in the i.p. and i.pl. fluids decreased rapidly after injection of hypotonic solution as compared with isotonic solution. The extent of this decrease was greater in the peritoneal cavity than in the pleural cavity. In the hypotonic condition, the area under the curve of concentration versus time (AUC) for platinum of i.pl. fluid was greater than that of i.p. fluid. When i.p. and i.pl. hypotonic CDDP were administered, the osmolarity of the fluid returned rapidly to the isotonic level, with equilibration in 30 or 180 minutes respectively. The lower osmolarity continued for a longer duration in the pleural cavity than in the peritoneal cavity. These results indicate that the pleural cavity may require a smaller amount of CDDP to achieve the same effect on intracellular uptake of platinum than the peritoneal cavity.

The therapeutic value of extensive gastric lymphadenectomy in gastric cancer is controversial. We retrospectively investigated the effect of extended lymphadenectomy on survival in 882 patients with gastric cancer.From 882 patients who underwent gastrectomy, D0 or D1 lymphadenectomy was performed on 137 patients, D2 lymphadenectomy on 524 and D3 lymphadenectomy on 221. Curative gastrectomy was performed on 771 patients and the 5-year survival rate of patients had undergone D0 or D1 lymphadenectomy (D1 group) and that of patients who had undergone D2 lymphadenectomy (D2 group) was compared with that of patients had undergone D3 lymphadenectomy (D3 group).In each stage, the 5-year survival rates of patients who had undergone curative operations (n = 771) were compared among the D1, D2 and D3 groups. The 5-year survival rates were as follows: Stage I: n = 510, 89.3% (D1 group: n = 91, 85.8%; D2 group: n = 384, 90.3%; D3 group: n = 35, 88.1%; p = 0.539), Stage II: n = 83, 77.8% (D2 group: n = 35, 82.7%; D3 group: n = 48, 74.3%; p = 0.601), Stage III: n = 133, 50.2% (D2 group: n = 46, 39%; D3 group: n = 87, 56.1%; p = 0.027), Stage IV: n = 45, 8.9% (D2 group: n = 10, 0%; D3 group: n = 35, 11.4%; p = 0.588). Postoperative complications were detected in 10.4% of the cases and the in-hospital mortality rate was 2.4%. The postoperative morbidity rate of the D3 group (15.8%) was significantly higher than that of the other groups (D1 group: 7.3% and D2 group: 9%; p = 0.008). However, the in-hospital mortality rate of the D3 group (1.4%) was not different from that of the other groups (D1 group: 3.7% and D2 group: 2.5%; p = 0.374).These results indicate that D3 lymphadenectomy might be performed as safely as D1 or D2 lymphadenectomy on patients with gastric cancer. In addition, D3 lymphadenectomy might provide a survival benefit for patients with Stage III or Stage IV gastric cancer.

Loss of heterozygosity (LOH) at 4 microsatellite loci (chromosomes 8p23, 8q24, 13q14 and 17p13) and the nuclear morphometric data were analyzed in 28 tumors from 28 patients with hepatocellular carcinoma (HCC). LOH was detected in 63.2% of the tumors with marker D8S264 (chromosome 8p23), in 52.6% with marker D8S1801 (chromosome 8q24), in 25.0% with marker D13S328 (chromosome 13q14), and in 52.4% with marker D17S796 (chromosome 17p13). LOH on 17p13 was most frequent in tumors that had reached more advanced stages (p0.05). The frequencies of LOH on 17p13 were significantly higher in tumors with portal invasion of cancer cells or intrahepatic metastasis than those without (p0.05). Moreover, capsule infiltration of cancer cells or hepatitis B viral infection were more frequently detected in patients with LOH on 17p13 in their tumors. The mean nuclear area (MNA) and the standard deviation (SDNA) of tumors with LOH on 17p13 were significantly larger than those of the other sites (p0.01). No significant relationships were found between the LOH on 8p23, 8q24 or 13q14 of HCC and the clinicopathologic data, MNA or SDNA. Therefore, LOH on 17p13 of HCC correlates with the stage, portal invasion of cancer cells, intrahepatic metastasis, and nuclear morphometry. These findings suggest that the LOH on 17p13 is closely connected to the progression of HCC and that nuclear morphometry is useful as an indicator of genetic alterations in HCC.

This interim analysis of the JFMTC study as of May, 1998 covers 321 gastrectomized patients with far-advanced stomach cancer from 135 institutions between November, 1993 and March, 1996. The intensive therapy group (I-group) received CDDP i.p. administration on resective surgery with 70 mg/m2 followed by CDDP i.v. of 80 mg/m2 (day 1, i.v.), accompanying 5-FU of 350 mg/m2/day (day 1-5, c.v.i.) in the 4th, 8th and 12th weeks. The I-group was randomly compared with the standard therapy group (S-group) of MMC of 6 mg/m2 i.v. in the 4th, 8th and 12th weeks and UFT of 3-4 capsules daily for postoperative one year. The results obtained were that 1. adverse reactions were found more in the I-group than in the S-group, particularly notable in the decrease in blood cells, loss of appetite and nausea/vomiting, and incidence of grade 3 or more being 13% (neutrophile leukocytes), 26% and 21%, respectively; 2. there was no significant difference between I- and S-groups in terms of 3-year survival or disease-free survival rates. (JFMTC: Japanese Foundation of Multidisciplinary Treatment for Cancer).

This interim report, for findings as of May, 1998, covers data on 435 gastrectomized patients with semi-advanced stomach cancer collected from 144 institutions between November, 1993 and March, 1996. The active arm of the study involved CDDP i.p. administration of 70 mg/m2 at the time of resective surgery, followed by UFT oral administration for one year at 3-4 capsules daily. A randomized control involved no adjuvant therapy after CDDP i.p. administered as in the active arm. The results obtained indicated no significant difference between the groups in terms of 3 year survival or disease free survival rates. Reports appearing elsewhere have suggested that 3-4 capsules/day of UFT may be insufficient to reach the threshold of the effective tissue level, and that 6 capsules may be necessary to obtain the expected results. (JFMTC: Japanese Foundation of Multidisciplinary Treatment for Cancer).

The presence of serum anti-p53 antibody has been reported to be associated with survival of patients with breast cancer, ovarian cancer, and hepatocellular carcinoma. To clarify prognostic significance of p53 antibody in colorectal cancer, serum p53 antibody was measured in patients with colorectal cancer. The 89 patients included 71 with colorectal cancer and 18 with colon polyp. An enzyme-linked immunosorbent assay was used to detect p53 antibodies in serum. Clinicopathological parameters such as age, sex, degree of differentiation of cancer, location of tumor, liver metastasis, stage classification, Dukes classification, CEA, CA19-9, and immunostaining of p53 and anti-p53 antibody were evaluated as prognostic factors of colorectal cancer. p53 antibody was positive in 18 of 71 (25%) with colorectal cancer, whereas it was positive in only 1 of 18 (6%) with colon polyp. The patients with p53 antibody had higher CEA and CA19-9 levels, higher positive rates of p53 protein expression in cancer cells, and higher liver metastasis rates. The p53 antibody positivity at stage classification I-IIIb/ Dukes classification A-C was significantly lower than that at stage classification IV/Dukes classification D. Overall survival in colorectal cancer patients with p53 antibody was significantly shorter than in those without p53 antibody. A Cox regression analysis showed that liver metastasis, stage classification, Dukes classification, CA19-9, and p53 antibody were significant prognostic factors in colorectal cancer. Serum anti-p53 antibody could serve as one of the prognostic factors in patients with colorectal cancer.

It has been recently reported that thymidine phosphorylase (dThdPase) is identical to platelet-derived endothelial cell growth factor (PD-ECGF). dThdPase is known to be one of the angiogenic factors. Expression of dThdPase has been detected both in cancer cells and in stroma cells. However, the role of dThdPase, in cancer cells and in stroma cells regarding, neovascularization in gastric adenocarcinoma is still unclear. In this study, we analyzed the correlation between dThdPase activity and neovascularization in gastric cancer.We obtained fresh samples (tumors and adjacent normal mucosas) from 67 patients with gastric cancer who underwent gastrectomy between 1993 and 1994. The dThdPase activity was analyzed by the ELISA (enzyme-linked immunosorbent assay) method and by the immunohistochemical staining. Microvessel density (MVD) and vascular endothelial growth factor expression of tumors were analyzed by immunohistochemical staining.The dThdPase activity of tumors (117 +/- 98 U/mg protein) was significantly higher than that of normal mucosas (46 +/- 36 U/mg protein, P0.0001). The expression of dThdPase was detected by immunostaining in stroma cells but not in mucosal cells in normal tissues. Based on the immunostaining results, 16 tumors expressed dThdPase both in cancer cells and stroma cells. The mean dThdPase activity and the mean MVD of these 16 tumors were significantly higher than those of 15 tumors with expression of dThdPase only in the stroma cells, and than those of 36 tumors with expression of dThdPase in neither the cancer cells or the stroma cells. The MVD of tumors strongly correlated with the dThdPase activity of tumors detected by the ELISA method (P0.0001).The total amount of dThdPase produced by cancer cells and stroma cells may influence tumor angiogenesis in gastric cancer.

Transforming growth factors beta (TGFs beta) are involved in a variety of important cellular functions, including cell growth and differentiation, adhesion, migration, extracellular matrix formation, and immune function. Moreover, it has been reported that TGFs beta are correlated with angiogenesis. However, the role of TGF-beta as an angiogenic factor in gastric carcinoma is still unclear.TGF-beta1 expression was determined in 101 patients with gastric carcinoma by immunohistochemical procedures, and this expression was compared in the current study with both the expression of vascular endothelial growth factor (VEGF), which is thought to be the most potent angiogenic factor, and microvessel density, to evaluate the effect of TGF-beta1 on the angiogenesis of gastric carcinoma tissues.TGF-beta1 expression was detected in 23 tumors (22.8%). TGF-beta1 expression was more frequent in differentiated than in undifferentiated gastric carcinoma. Furthermore, TGF-beta1 expression was significantly correlated with the depth of invasion and the stage of disease. There was a close correlation between TGF-beta1 expression and VEGF expression. There was no correlation between TGF-beta1 expression and microvessel density, whereas VEGF expression was significantly correlated with microvessel density. With regard to prognosis, the 5-year survival rate was 55.9% for patients with TGF-beta1 positive tumors and 67.0% in patients with TGF-beta1 negative tumors. Accordingly, the prognosis for patients with TGF-beta1 negative tumors was significantly better than that for patients with TGF-beta1 positive tumors. Multivariate analysis indicated that lymph node metastasis, tumor size, and TGF-beta1 expression were independent prognostic factors.These results suggest that TGF-beta1 might be associated with tumor progression by indirectly stimulating angiogenesis through the up-regulation of VEGF expression in gastric carcinoma.

When the DNA of cells is damaged, wild-type p53 protein induces the expression of p21 (Waf1/Cip1/Sdi1), and regulates the progression of the cell cycle by inducing G1 arrest. Thus, wild type p53 or p21 protein negatively regulates cancer cell proliferation. However, in tumor with loss of expression of functional wild-type p53 protein by mutation or allelic deletion of the gene for p53, whether the proliferative activity of cancer cells might be accelerated or not is unclear. In this study, we investigated the correlation between the level of expression of mutated p53 protein and the proliferative activity of cancer cells in advanced gastric cancer.Ninety-seven samples from patients with gastric cancer that had invaded the serosa without lymph node metastasis (t3, n0, stage II) were investigated by immunohistochemical staining with a monoclonal antibody against p53 and against p21, and with the monoclonal antibody Ki-67. DNA ploidy patterns were analyzed by flow cytometry. The immunoreactivity against p53 and the proliferative activity of cancer cells were scored in terms of a labeling index (LI; percentage of immunostained cells) in each case. Moreover, the prognostic values for 93 surviving patients were evaluated by univariate and multivariate analysis.The mean p53 LI was 24% (range: 0-82.4%) and the mean Ki-67 LI was 23.1% (range: 0-70.7%) in 97 tumors. The expression of p21 protein was detected in 30 of 97 tumors (30.9%) and DNA aneuploidy was detected in 36 of 97 tumors (37.1%). There was significant correlation between the p53 LI and the Ki-67 LI (r = 0.61, t = 7.456, p0.001) in 97 tumors. Although, no significant difference was detected, the mean p53 LI (18.3%) of 30 tumors with expression of p21 protein was lower than that of 67 tumors without expression of p21 protein (26.6%, p = 0.096). However, no significant correlation between expression of p21 protein and Ki-67 LI was observed. The p53 LI was not an independent prognostic factor in 93 surviving patients by multivariate survival analysis (p = 0.069). However, the 5-year survival rate of 50 patients with a low level of p53 LI (p53 LI (or = 10%, 78.3%) was significantly better than that of 43 patients with a high level of p53 LI (p53 LI10%, 62.1%, p = 0.045).Accumulation of mutated p53 protein might suppress the expression of p21 protein in gastric adenocarcinoma, and cancer cells with overexpression of mutated p53 protein might have a high proliferative activity.

Of patients radically operated on for breast cancer in our department, 46 patients who had recurrent breast cancer were clinicopathologically evaluated to clarify the prognostic factors of recurrent breast cancer. Furthermore, p53, p21 and cyclin D1 protein expression were studied immunohistochemically and their prognostic value was evaluated. The relapse-free interval was highly related to the survival rate after the recurrence. p53 overexpression was correlated with the progression of clinical stage and lymph node metastasis and negatively correlated with the expression of estrogen receptor. The p21 positive and p53 negative cases had a significantly better prognosis than the p21 negative and p53 positive cases. The combination of p53 and p21 protein expression seemed to have prognostic value in recurrent breast cancer.

The human MAGE gene products are recognized by major histocompatibility complex-restricted cytotoxic T lymphocytes. We analyzed by RT-PCR the expression of MAGE-1, MAGE-2, MAGE-3 and HLA-A2 genes in 10 human gastric cancer cell lines and 46 human stomachs removed due to advanced gastric carcinomas. All the cell lines expressed MAGE genes, except for MKN-45 and -74 which lacked the expression of MAGE-1 and -3 genes in this study. Of the 46 gastric carcinomas, MAGE-1, -2 and -3 genes were expressed in 14 (30%), 10 (22%) and 26 (57%) cases, respectively, regardless of histological type. Normal gastric mucosa and intestinal metaplastic mucosa showed no expression of these genes. HLA-A2 gene expression was noted in 14 both normal and carcinoma cases. Simultaneous expression of MAGE-3 and HLA-A2 genes was noted in 7 cases. Mean apoptotic and Ki-67-labeling indices (AI and KI) of carcinoma cells were 2.3 +/- 0.5 and 48.1 +/- 6.0 in 7 cases, and 2.8 +/- 0.2 and 47.3 +/- 2.7 in the other 39 cases lacking the expression of MAGE-3 and/or HLA-A2 genes, respectively. The two-year survival rate did not differ between the two groups. Although this study confirmed the relatively higher expression of the MAGE gene family in human advanced gastric carcinomas, it might suggest that simultaneous expression of MAGE-3 and HLA-A2 genes does not necessarily imply the induction of cancer cell apoptosis by CTL.

An examination was made of the postoperative long-term effects of limited gastrectomy (LG) on nutritional status, as one of the factors that influence quality of life.Nutritional status in 33 patients who underwent LG for early gastric cancer, was compared with that of 36 patients who underwent standard gastrectomy (SG). Nutritional conditions were assessed preoperatively, and then 1 and 2 weeks, 1 and 6 months, and 1, 2 and 3 years after surgery.Postoperative recovery of both body weight and body fat mass was greater in LG than SG, and there were significant differences between the two groups of patients from six months after surgery until three years had elapsed.LG minimized the extent of nutritional impairment during long-term recovery from surgery, as compared with SG. LG would seem to be a suitable operative procedure for some patients with mucosal gastric cancer, without reducing radicality for cancer.

Telomerase activity was analyzed in 52 patients with resected esophageal squamous cell carcinoma by telomeric repeat amplification protocol (TRAP) assay. Telomerase activity was detected in 41 out of 52 (79%) carcinomas and detected in 46 out of 52 (89%) normal esophageal epitheliums adjacent to the carcinomas. In carcinoma, the telomerase activity was detected more frequently in relatively early stages of carcinoma (in 8 of 9 patients with stage 0, in 3 of 3 patients with stage I, and in 4 of 4 patients with stage II). However, the telomerase activity was detected in only 8 of 12 patients with stage III and in 16 of 21 patients with stage IV. Thus, the percentage of patients who had tumors without telomerase activity increased in more advanced stages. These results indicate that telomerase activity may correlate with carcinogenasis of esophageal cancer, however, it may not correlate with progression of esophageal squamous cell carcinoma.

In this study, we investigated the correlation between the telomerase activity and the microsatellite instability in colorectal cancers and adenomas. Telomerase activity was detected in 75% of cancers and in 33% of adenomas. Microsatellite instability was detected in 24% of sporadic cancers, in 54% of multiple cancers, and in 22% of adenomas. Telomerase activity and microsatellite instability were independent events in colorectal carcinogenesis. The patients with telomerase activity and microsatellite instability showed poor prognosis. These results indicate that colorectal cancers with both telomerase activity and microsatellite instability may have more malignant potential, and adenomas with both them may be more intensive precursor of colorectal cancers.

To evaluate whether the expression of p53 and that of p21 are independent prognostic factors in patients with advanced gastric cancer, we investigated clinicopathological factors and the expression of p53 and p21 in 158 patients with gastric cancer that had invaded the serosa and who had undergone curative gastrectomy. In multivariate survival analysis of 156 surviving patients, we evaluated the size of the tumor, lymph node metastasis, venous invasion, lymph node dissection, expression of p53, and expression of p21 as independent prognostic factors. Moreover, we divided patients into four groups according to the expression of p53 and p21 in their tumors [group A, p53-/p21-, N = 40 (one died within 30 days of surgery); group B, p53-/p21+, N = 23; group C, p53+/p21-, N = 58; and group D, p53+/p21+, N = 37 (one died within 30 days of surgery)]. The 5- and 10-year survival rates of 39 patients in group A were 71.7% and 64.3%, those of 23 patients in group B were 81.4% and 81.4%, those of 58 patients in group C were 35.6% and 30.2%, and those of 36 patients in group D were 67.9% and 60.7%. The prognosis of patients in group C was poorer than that of patients in the other three groups. This result indicates that the evaluation of the expression of both p53 and p21 expression might provide prognostic information that is more accurate than that provided by evaluation of the expression of p53 alone.

We examined the relationship between apoptosis induced by 5-fluorouracil (5-FU) that was given preoperatively to colorectal cancer patients and DNA ploidy pattern, and investigated the cell cycle changes, and the expression of Ki-67. Twenty-nine patients with advanced colorectal cancer were divided into four groups, 3 days, 5 days, 7 days, and 10 days. Groups received continuous intravenous 5-FU at 500 mg/body/day preoperatively. Then, patients were divided into two groups by DNA ploidy pattern, diploid(D) and aneuploid(A). Apoptotic cells were stained by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method. The expression of Ki-67 was examined by immunohistochemical staining. We used flow cytometry (FCM) for analysis of cell cycle distribution. Apoptosis of cancer cells was mostly increased in 7 days 5-FU administration in both D and A groups. The expression of Ki-67 was reduced according to the prolongation of the term of 5-FU administration in both D and A groups. We assessed S-phase fraction (SPF) to evaluate the cell cycle changes by 5-FU. Tumor samples of all patients after injection of 5-FU showed S-phase accumulation. The ratio of SPF (after 5-FU/before 5-FU) was the highest in the 5-day 5-FU administration group in both D and A groups. We concluded that apoptosis and S-phase accumulation were increased, and proliferative activity was decreased by preoperative 5-FU administration in colorectal cancer patients. However, there was no clear correlation between DNA ploidy pattern and these changes.

Many genes participate in the regulation of cell cycle progression from G1 to S phase. Functional loss of one or more of these genes has been reported to be associated with carcinogenesis and/or tumor progression and poor prognosis in many cancers. In a series of 126 patients with hepatocellular carcinoma (HCC), we immunohistochemically evaluated tumor expression of the cell cycle-related gene protein products of Rb, p21 (WAF1), and p53. Positive immunostaining for Rb, p21, and mutant p53 protein was detected in 58%, 33%, and 37% of the tumors, respectively. The proportion of HCCs exhibiting aberrant p53 protein expression increased significantly with advancing stage of disease (p0.001), poorer histological classification of differentiation (p0.01), and increasing tumor size (p0.01). A decrease in the proportion of HCCs expressing p21 protein was also associated with advancing clinical stage of disease (p0.01), and larger tumor size (p0.05). The only clinicopathological feature found to be associated with Rb status, was intrahepatic metastasis, which occurred with a higher frequency in HCCs exhibiting positive immunoreactivity for Rb protein expression (p0.05). Multivariate survival analysis revealed that, amongst the protein products of the different genes evaluated, only positive immunostaining for aberrant p53 protein expression served as an independent prognostic indicator, being significantly associated with worse survival in patients with HCC (p = 0.023). Analysis for relationships between gene products showed an inverse correlation between expression of aberrant p53 protein and p21 protein (p0.01), and also an inverse correlation between p21 protein and Rb protein expression (p0.05) in these cases of HCC. These findings demonstrate that positive immunostaining for mutant p53 protein expression is a significant indicator of tumor progression and poor prognosis, confirm that p21 protein expression is induced in a p53-dependent manner, and suggest that Rb protein expression may be regulated to some extent by p21 in HCC.

The aim of this study was to investigate the relationship between the expression of p53 protein and radiochemosensitivity in patients with advanced esophageal cancer. We analyzed the expression of p53 protein by an immunohistochemical method in tumors from 28 patients with advanced esophageal cancer who had been treated by absolute noncurative resection. Four patients died from postoperative complications. Fourteen of the remaining 24 patients were treated with postoperative radiochemotherapy. The expression of p53 protein was detected in tumors from 16 of 28 patients (57.1%). The mean survival period was 16.4 months for 14 patients treated with radiochemotherapy, as compared to 6.9 months for 10 patients who were not treated with radiation therapy (P = 0.112). In the case of the 14 patients treated with radiochemotherapy, the mean survival period was 24.7 months for seven patients with p53-negative tumors while it was only 8 months for seven patients with p53-positive tumors. However, difference was not statistically significant (P = 0.149). Postoperative radiochemotherapy prolonged the survival of some patients with noncuratively operated esophageal tumors that were p53-negative. However, the enhanced survival was not statistically significant.

Two colon preparation methods were compared for their efficacy. The comparison was carried out between whole bowel irrigation using polyethylene glycol-electrolyte lavage solution (PEG-ELS) and total parenteral nutrition (TPN).Twenty-six consecutive patients suffering from colorectal cancer without apparent obstruction were randomly divided into two groups. Twelve patients received 2 litres of PEG-ELS one day before the operation following two tablets of sennoside (24 mg) two days before the operation without restriction of food until PEG-ELS. Fourteen patients received magnesium citrate (34 g) and sennoside (24 mg) on the day before the operation with 7 days' TPN and fasting.The average time of defecation was 9 in the PEG-ELS group and 5 in the TPN group (p0.001), with little difference between the necessary times to complete defecation (PEG-ELS: 280 min. vs. TPN: 260 min.). However, although half of the PEG-ELS group complained of side effects of abdominal fullness the overall incidence of side effects in both groups were the same of 50%. There were no significant changes between the data of blood test and routine chemistry before and after the bowel preparation in both groups. The purity of the colorectum judged at operation in both groups were excellent, no residue at all, except in one patient of the TPN group. The total bacterial counts of the rectal samples obtained at operation of PEG-ELS group were fewer than that of TPN group but not statistically significant.Colon preparation using PEG-ELS is just as good as the 7-days' TPN treatment and fasting, and exceeds from the standpoint of saving time.

To estimate the correlation between the expression of mutated p53 protein and the nuclear DNA ploidy of gastric cancer cells, as well as the influence of these factors on the prognosis of patients with advanced gastric cancer, we selected 161 patients with gastric cancer that had invaded the serosa and who were treated by potentially curative gastrectomy. The expression of p53 protein was detected in 95 (59%) patients and aneuploidy was detected in 70 (43.5%) patients. The percentage of tumors with DNA aneuploidy among the 95 tumors that were p53-positive was 51.6% and this percentage was significantly higher than that of tumors with DNA aneuploidy among the 66 tumors that were p53-negative (31.8%, P = 0.02). The percentage of cells in the G2M phase of the cell cycle was significantly higher in p53-positive tumors than that in p53-negative tumors (P = 0.024). The 161 patients were divided into four groups according to the expression of p53 protein and the DNA ploidy of their tumors. Patients with p53-positive tumors and DNA aneuploidy were allocated to group A (n = 49), patients with p53-positive tumors and DNA diploidy were allocated to group B (n = 46), patients with p53-negative tumors and DNA aneuploidy were allocated to group C (n = 21), and patients with p53-negative tumors and DNA diploidy were allocated to group D (n = 45). The 5-year survival rate of group D was 73.7% and it was significantly higher than that of group A (41.8%, P = 0.007). These observations indicate that mutated p53 protein might accelerate cell proliferation. Moreover, analysis of both p53 status and DNA ploidy of tumors seems to provide very important information for evaluation of the prognosis of patients with advanced gastric cancer.

A retrospective study was designed to evaluate the clinical significance of preoperative serum levels of carcino-embryonic antigen (CEA) and CA72-4 in patients with advanced gastric adenocarcinoma.Pre-operative serum levels of CEA and CA72-4 were analyzed and compared with the proliferative activity of cancer cells, monitored in terms of the number of argyrophilic nucleolar organizer regions (AgNORs), in tumors from 153 patients with resected gastric adenocarcinoma with serosal invasion.Elevated levels of CEA were frequently found in patients with lymph node metastasis and liver metastasis. By contrast, elevated levels of CA72-4 were frequently found in patients with peritoneal metastasis at the time of operation. A close correlation was found between pre-operative levels of serum CA72-4 and the AgNOR scores of tumors. Even when apparently curative operations were performed, peritoneal metastasis occurred these were detected quite soon, after apparently curative operations in patients with high pre-operative serum levels of CA72-4 and tumors with high proliferative activity.This phenomenon might explain the poor prognosis of patients with high levels of CA72-4.

The relationship between the expression of p53 and p21 in cases of advanced gastric cancer (t3) with lymph node metastasis [n1(+) or n2 (+)] and the long-term effects of continuous hyperthermic peritoneal perfusion (CHPP) immediately after resection was evaluated.Between 1983 and 1989, 74 patients with gastric cancer [t3, n1(+) or n2(+)] underwent curative gastrectomy and survived without postoperative complications. These patients were followed up for at least six years.Thirty-seven patients died from recurrence of cancer. In 34 patients who had tumors designated p53-/p21+ or p53+/p21+, CHPP reduced the percentage of patients who died from recurrence of cancer from 10/20 (50%) to 3/14 (21.4%, P = 0.184). However, in 40 patients who had tumors designated p53-/p21- or p53+/p21-, the percentage of patients who were treated with or without CHPP and died from recurrence of cancer was the same, 6/10 (60%) and 18/30 (60%). In the case of patients who had p21-positive tumors, the 5-year survival rate of 11 patients who were treated with CHPP (72.7%) was higher than that of 17 patients who were not treated with CHPP (41.2%, p = 0.027). However, in the case of patients with p21-negative tumors, the 5-year survival rate of 10 patients who were treated with CHPP (50%) was not very different from that of 28 patients who were not treated with CHPP (42.9%, p = 0.308).These results indicate that the CHPP might be effective in preventing the recurrence of cancer in patients with p21-positive regardless of the expression of p53.

We present a method for the cytological evaluation of the therapeutic effects of various treatments on unresectable metastasis in the liver from colorectal cancer. A degeneration index (DI) for cancer cells obtained by aspiration was determined in 17 patients before and after treatment. The relationships of the rate of tumor reduction and of the post-treatment survival period to the treatment-induced changes in DI were studied. The treatment-induced change in DI was represented by the difference between the pretreatment DI and the posttreatment DI. The treatment-induced change in DI was significantly correlated with the rate of tumor reduction (r = 0.794, P0.001). The survival rate was significantly higher in patients with higher changes in DI than in patients with lower changes (P0.05). The change in the DI of cancer cells seems to be a useful parameter for evaluating the effects of chemotherapy on unresectable liver metastases from colorectal cancer.

To investigate the prognostic significance of DNA ploidy pattern and DNA index (DI), DNA contents were measured by flow cytometer in 412 patients with colorectal cancer and correlation between their prognoses and DNA contents were analyzed on the same clinical stage. There were significant differences in the survival rate and the incidence of tumor recurrence between diploid and aneuploid tumors, especially the poor survival rate and frequent tumor recurrence in the aneuploid tumor with DI above 1.5. Cox's multiple regression proportional hazard model was used to investigate the prognostic value of DNA ploidy pattern, DI and clinicopathological findings. From these analyses, DI 1.5 was found to be the most significant prognostic factor. These results suggest that flow cytometrically evaluated DI values have a relevant independent power for predicting the clinical outcome of colorectal cancer patients.

This study examined p53 gene alterations in human gastric mucosa, intestinal metaplasia and well-differentiated (cohesive type) adenocarcinomas to clarify to the role of the p53 gene in gastric tumorigenesis by means of fluorescence in situ hybridization (FISH), polymerase-chain-reaction-single-strand-conformation polymorphism (PCR-SSCP) and immunohistochemistry. Gene alterations were compared with numerical changes of chromosome 17. Samples were obtained from 31 surgically resected stomachs affected with gastric cancer. There was no nuclear p53 protein in cells from normal gastric mucosa. Among 23 specimens of intestinal metaplasia, cells with p53 protein were variably detected in 5 incomplete metaplasias (colonic type). A histological study revealed a mildly dysplastic appearance. PCR-SSCP identified p53-gene mutation in exons 5 and 8 in 2 of the 5 samples. Numerical aberrations of chromosome 17 and the p53 gene were undetectable both in normal mucosa and in intestinal metaplasia. Variable numbers of tumor cells contained p53 protein in 13 (54%) of 24 carcinomas, and PCR-SSCP revealed abnormal bands in 5 of them. Mutations were detected in exons 5, 7, 7, 7 and 8. FISH analysis demonstrated that 6 carcinomas emitted 3 or 4 signals for chromosome 17, and 7 gave one signal for the p53 gene in over 20% of the cells. Ten (77%) of 13 carcinomas examined by FISH appeared to show a p53-gene deletion.

Between 1976 and 1984, 234 patients with advanced gastric cancer (pT3, pT4) were curatively gastrectomized and intraperitoneal free cancer cells which were detected by lavave of the Douglas cavity were recognized in 17.8% of the patients with pT3 and 25.0% of those with pT4. Even if curative surgery was performed for patients with free cancer cells, their five-year survival rate was very poor. The five-year survival rate was very poor. The five-year survival of patients with curatively operated gastric cancer without free cancer cells was 49.3%, whereas that of patients with free cancer cells was significantly lower (15.4%). Then the results of treatment for patients with free cancer cells were analyzed in regard to the effect of continuous hyperthermic peritoneal perfusion (CHPP) as prophylaxis of peritoneal metastasis. Five-year survival rate in the patients treated with CHPP was 33.3% and it was only 4.2% in the controls. In patients without intraperitoneal free cancer cells, however, no difference was observed in the five-year survival rates between those patients who had been treated with CHPP and the controls. Therefore, it is reasonable to assume that CHPP can be effective in preventing peritoneal recurrence of resected gastric cancer that is positive for free cancer cells in the peritoneal cavity and peritoneal micrometastasis.

The proliferative activity of cancer cells, as indicated by numbers of argyrophilic nucleolar organizer regions (AgNORs), at five sites along the invasive margin of the tumor was assessed for 65 patients with histologically proven and curatively operated gastric adenocarcinoma that had invaded the serosa. The mean number of AgNORs in cells at the invasive margin of the tumor was 4.6. Patients with high AgNOR scores (AgNOR scoreor = 4.6, n = 34) had a significantly elevated frequency of lymphatic involvement (82%) as compared with patients with low AgNOR scores (AgNOR scores4.6, n = 31, 45%), regardless of the size and histologic type of the tumor, and the depth of invasion of the gastric wall. The 5-year survival rate of 33 patients with high AgNOR scores (22.7%) was significantly poorer than that of 31 patients with low AgNOR scores (55.4%, p = 0.024). Thus, the AgNOR score of cancer cells at the invasive margin of the tumor appears to be a good predictor of the prognosis of patients with advanced gastric cancer with serosal invasion.