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Oxidative stress plays a crucial role in Alzheimer’s disease (AD) from its prodromal stage of mild cognitive impairment. There is an interplay between oxidative stress and the amyloid β (Aβ) cascade via various mechanisms including mitochondrial dysfunction, lipid peroxidation, protein oxidation, glycoxidation, deoxyribonucleotide acid damage, altered antioxidant defense, impaired amyloid clearance, inflammation and chronic cerebral hypoperfusion. Based on findings that indicate that oxidative stress plays a major role in AD, oxidative stress has been considered as a therapeutic target of AD. In spite of favorable preclinical study outcomes, previous antioxidative components, including a single antioxidative supplement such as vitamin C, vitamin E or their mixtures, did not clearly show any therapeutic effect on cognitive decline in AD. However, novel antioxidative supplements can be beneficial for AD patients. In this review, we summarize the interplay between oxidative stress and the Aβ cascade, and introduce novel antioxidative supplements expected to prevent cognitive decline in AD.

Oxidative stress is part of the entire pathological process that underlies the development of Alzheimer's disease (AD), including the mild cognitive impairment (MCI) stage. Twendee X (TwX) is a supplement containing a strong antioxidative mix of eight antioxidants, which has been shown to have a clinical and therapeutic benefit in AD model mice. Here, we conducted a multicenter, randomized, double-blind, and placebo-controlled prospective interventional study to evaluate the efficacy of TwX in mitigating MCI. The primary outcomes were differences in Mini-Mental State Examination (MMSE) and Hasegawa Dementia Scale-revised (HDS-R) scores between baseline and six months for placebo and TwX groups. Seventy-eight subjects with MCI were randomized into placebo (n = 37) and TwX (n = 41) groups. MMSE scores at six months differed significantly between the TwX and placebo groups (p = 0.018), and HDS-R scores for the TwX group exhibited a significant improvement at six months relative to baseline (p = 0.025). The TwX group did not show any change in affective or activities of daily living scores at six months. The present study indicates that strong antioxidative supplement TwX is clinical beneficial for cognitive function in subjects with MCI.

[Purposes of the study] Reactive oxygen species (ROS) are associated with clinical condition of cancer patient, and closely involved in cancer growth and metastasis. Twendee X (TWX) is a composition consisting of Vitamin C, Glutamine, Cystine, Riboflavin, Succinic acid, Fumaric acid, Coenzyme Q10, and Niacin. TWX strongly reduces oxygen radicals or ROS in vitro and in vivo (World Intellectual Property Organization WO2013/072441 A1 COMPOSITION FOR PROTECTION AGAINST CELL-DAMAGING EFFECTS). TWX has significant effects of reducing tumor growth and inhibition metastasis. Hydrogen peroxide level (d-ROMs test) of tumor bearing nude mouse in serum was significantly reduced and increased NK-activity (2016 AACR meeting). Mechanism of action was examined for TWX using hepatic cancer cell line HepG2. [Experimental procedures] Human hepatic cancer cell line HepG2 was used in this study. Cell proliferation experiments of antioxidant by ICDD (France). Metabolome analysis was by Human Metabolome Technologies (Tokyo). In both experiments, HepG2 cell line treated one hour with TWX (60ug/ml). [Data] TWX inhibits the RAF-MEK-ERK pathway starting from ERK. Metabolome analysis shows total shift of metabolism, and especially increase mitochondrial metabolism and ATP production. [Conclusion] Effects of TWX on cancer cell line are not only increase immune activity, but also inhibit proliferation pathway, and mitochondrial metabolism change. Several publications using Vitamin-E or Cysteine as anti-oxidant showed cancer progression or increase metastasis. However, Vitamin-E or Cysteine has limited anti-oxidant effects and hydrogen peroxide level (d-ROMS test) dose not change. On the other hand, TWX has significant effects reduce hydrogen peroxide level, and Japanese Association for Dementia Prevention already started clinical trial for MCI and after stroke patients. For the cancer patients, clinical trial using TWX combination with standard therapy will be start soon. Citation Format: Haruhiko Inufusa. Mechanism of action for antioxidant Twendee X on hepatic cancer cell line HepG2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3490.

Previously we showed that MRP-1/CD9 might prevent tumor metastasis by suppression of cell motility and invasion of tissue barriers. The present study explored the possibility of preventing metastasis of mouse melanoma BL6 by expression of MRP-1/CD9 through gene transfer. A replication-deficient adenovirus vector was used for the in vivo transfer of MRP-1/CD9 cDNA. Intratumor injection of an adenovirus vector (rAd-MRP-1/CD9) expressing MRP-1/CD9 resulted in a 73.7% reduction in the number of pulmonary metastases of mice and the median survival time of mice treated with rAd-MRP-1/CD9 was significantly longer than those treated with the rAd-beta-gal vector (103.2 approximately plus;8.5 days vs 71.2 approximately plus;5.2 days, P0.001 respectively). These results support the expression of MRP-1/CD9 through gene transfer as a therapeutic strategy for preventing metastases and prolonging survival, and support the feasibility of gene transfer in a clinically relevant setting.

BACKGROUND Tumor procoagulant is one of the factors responsible for disseminated intravascular coagulation and metastasis. The authors found procoagulant activity in LK52 human squamous cell carcinoma cells, which they designated cancer cell-derived blood coagulating activity 1 (CCA-1). A monoclonal antibody (MoAb) was generated to characterize this CCA-1 procoagulant activity. To date, antibodies that show an inhibitory effect on procoagulant activity as well as high reactivity in cancer cells are well known for their tissue factor specificity. METHODS Characterization of the procoagulant activity of CCA-1 was performed and an anti-CCA-1 MoAb, FS01, was generated. CCA-1 expression on the cancer cell surface was examined by flow cytometry. Procoagulant activity of various cancer cell lines and the inhibitory effect of the FS01 MoAb on this procoagulant activity was monitored by a clot timer. RESULTS The enzymologic character differed from that of cancer procoagulant (CP). The FS01 MoAb inhibited the procoagulant activity of CCA-1, but did not inhibit that of tissue factor. A positive correlation was observed between the expression intensity of CCA-1 and the inhibitory effect of the FS01 MoAb on the procoagulant activity of cancer cell lines. Expression of CCA-l was observed more frequently than that of tissue factor in human cancer cell lines. CONCLUSIONS The FS01 MoAb generated in the current study is a new antibody that reacts with various cancer cell lines, but not with normal cells. FS01 inhibits cancer cell-derived procoagulant activity and does not react with tissue factor and CP. CCA-1, which is recognized by the FS01 MoAb, appears to play a major role in cancer cell-derived procoagulant activity. Cancer 1998;82:1563-9. © 1998 American Cancer Society.

We have generated a monoclonal antibody (MAb), FS01, which inhibits the procoagulant activity (CCA-1) produced by a human squamous cell carcinoma cell line, LK52. Expression of the antigen recognized by FS01 MAb in various cancer cell lines correlated well with the procoagulant activities of the expressing cell lines. Our objective was to characterize the molecule reacting with FS01 MAb and to analyze its involvement in the CCA-1 procoagulant activity. The molecule was identified as a glycolipid and found to be involved in the procoagulant activity because both procoagulant activity and reactivity to FS01 MAb were lost after endoglycoceramidase treatment of CCA-1. Furthermore, FS01 MAb recognized the Lewis Y (Le y ) antigen. To confirm the involvement of a glycolipid incorporating the Ley antigen in the procoagulant activity, we attempted to purify CCA-1 from LK52 culture supernatant. In one of the purification steps, a fraction containing low procoagulant activity (CCA-1p) separated from the Le y -positive fraction (CCA-1c). Although CCA-1c alone did not show procoagulant activity, the procoagulant activity of CCA-1p was augmented by CCA-1c and this augmentation was inhibited by FS01 MAb. Furthermore, CCA-1c enhanced the procoagulant activity of 33 cell lines tested as well as CCA-1p. In addition, purified Ley glycolipid from canine intestine augmented the procoagulant activity of CCA-1p, and this augmentation also could be inhibited by FS01 MAb. We conclude that Ley glycolipid is a co-factor for the procoagulant activity derived from cancer cells. Int. J. Cancer 73:903‐909, 1997. r 1997 Wiley-Liss, Inc.

A case of gas-producing liver abscess complicated with diabetes mellitus was reported. A 66-year-old woman was admitted to the hospital because of a high fever and abdominal pain in the right upper quadrant. She showed a stinking abdominal pain and Blumberg's sign, complicated with shock symptoms 3 days after the admission. Emergency operation was performed under a diagnosis of peritonitis and septic shock due to rupture of liver abscess. Operative gross finding disclosed no pus nor ascites in the abdomen. An echo free space was found in the right lobe of the liver by ultra sound examination. The echo free space of the liver was punctured, and only gas with feces's smell was discharged. Therefore, silicon pleated drain was inserted into the cavity of the liver. Klebsiella pneumoniae and E. coli were detected from the drain discharge and arterial blood. The patient was recovered from disseminated intra-vascular coagulation and adult respiratory distress syndrome which occurred after the operation. Only 29 cases of gas-producing liver abscess have been reported in Japan. Twenty three (79%) of these 29 cases were associated with diabetes mellitus. It is believed that the disease is rare and has a poor prognosis. This is a case that appropriate surgical treatment can survive such patient's life.

Background. Expression of oncofetal fibronectin (oncFN) and normal fibronectin (norFN) in colorectal cancer specimens was examined to investigate the correlation between fibronectin localization and histologic grade, liver metastasis, and prognosis. Methods. Immunohistochemical staining of oncFN and norFN was performed on 99 primary lesions and 12 liver metastases of colorectal cancer. The expression of norFN and oncFN was evaluated by grading the intensity of staining as negative, positive, or strongly positive. Results. Positive staining for oncFN correlated positively with increasing stage. The rate of strongly positive staining for oncFN was 53% for primary lesions with liver metastasis, significantly higher than the oncFN-positive rate of 13% for metastasis free cases (P < 0.05). Liver lesions had an oncFN-positive rate of 92%. The postoperative 5-year survival rate for 51 cases classified as Dukes Stage C was 77.8% for oncFN-negative cases, 36.5% for oncFN-positive cases, and 22.2% for oncFN-strongly positive cases; these rates were significantly different (P < 0.01). Conversely, there was no correlation between norFN and any clinical variable. Conclusion. Expression of oncFN is correlated with a poor prognosis of Dukes C colorectal cancer and may serve as a useful postoperative prognostic sign. Cancer 1995;75:2802–8.

[Purposes of the study] Reactive oxygen species (ROS) are associated with clinical condition, and closely involved in cancer growth and metastasis. Twendee X is a composition consisting of Vitamin C, Glutamine, Cystine, Riboflavin, Succinic acid, Fumaric acid, Coenzyme Q10, and Niacin. Twendee X strongly reduces oxygen radicals or ROS in vitro and in vivo (World Intellectual Property Organization WO/2013/072441 COMPOSITION FOR PROTECTION AGAINST CELL-DAMAGING EFFECTS). Effects of Twendee X on human colon cancer cell lines growth and metastasis in nude mouse was examined. [Experimental procedures] Human colon cancer cell line RPMI4788 human gastric cancer cell line MKN45 was used in this study. Ten nude mice (BALB/C nu-nu, 4wks, Male) per each group of experiments were used, and repeated twice. Two cell lines were inoculated in subcutaneous of nude mouse, and tumor growth monitored for days. Lung metastasis was counted weeks after injection RPMI4788 cells into the tale vein of nude mouse. Twendee X was added to the free drink water around 37mg/kg/day by calculation. Serum of tumor bearing nude mouse was sampled just before sacrifice. Free Calpe Diem (Diacron International, Italy) was used to measure ROS related marker d-ROMs test (detect hydrogen peroxide). Natural Killer cell (NK) activity also measured in tumor bearing nude mouse. [Data] Tumor growth of RPMI4788 and MKN45 was reduced 25% and 40% respectively by Twendee X administration. Metastatic nodule of RPMI4788 was reduced to 1/7 by Twendee X administration. Increase value of d-ROMs test reduced to 60% compare to the control by Twendee X administration. NK activity of RPMI4788 tumor bearing nude mouse was reduced to 28% compare to the normal mouse, and increased to 56% by Twendee X administration. [Conclusion] Twendee X has significant effects of reducing tumor growth and inhibition metastasis. Hydrogen peroxide level (d-ROMs test) of tumor bearing nude mouse in serum was significantly reduced. ROS effects on immune cell activity, and Twendee X recovered suppressed NK activity. Thus, Twendee X effects on ROS inhibition and suppressed RPMI 4788 tumor growth and metastasis to the lung of nude mouse. Metastatic nodules receiving larger blood flow compare to the tumor in the back of mouse, thus means anti-tumor immune cells in blood may easier attack cancer cells. Citation Format: Haruhiko Inufusa. Targeting reactive oxygen species in human cancer cell growth and metastasis in nude mouse. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2812.

大腸癌および大腸癌肝転移で切除された原発巣99例と肝転移巣12例について癌胎児性fibronectin (onc FN) および正常型fibronectin (nor FN) の免疫組織化学的染色を行い, oncFNとnor FNの局在と進行度, 肝転移および予後との関連を検討した.染色性を陰性 (-), 陽性 (+), 強陽性 (++) と3群に分け判定した.onc FNの発現形式はnorFNと異なり, carcinoembryonic antigen (CEA) などの分泌型抗原の様相を呈した.oncFN染色性は, stageの進行と正の相関を認めた.また肝転移例原発巣のonc FNの強陽性率は53%であり, 非肝転移例の13%と比較し有意に高率であった (p

Two monoclonal antibodies AI18 (IgM) and AI51 (IgG1) were generated against the Factor X activating coagulant serine protease which extracted from LK52 human squamous cell carcinoma cell line. Both antibodies inhibit the activity of LK52 coagulant serine protease, but not of Russel's viper venom. AI18 and AI51 react with human lung and colon cancer cell lines, but not react with mouse cancer cell lines, rabbit cancer cell line or human fibroblast cell lines from lung and colon. From these results, LK52 coagulant serine protease was named as coagulant cancer antigen 1 (CCA-1). This is a first report of a novel CCA-1 expression on various human cancer cell lines. CCA-1 may play a important role in unbalanced haemostasis of cancer patients.

新しい腺癌肝転移モデルを樹立した.1985年10月に当院で手術した直腸癌組織をヌ-ドマウスの背部に移植し, ヒト大腸癌細胞株KUM・RK-4Nを樹立し以後継代した-1990年2月にこの腫瘍より初代培養を行い培養細胞株を樹立した.この細胞をヌードマウスに尾静脈注入および前腸間膜静脈より注入したところ, 約2-3週間で肺および肝臓に明瞭な結節型の転移巣を生じた.ヒト癌でのヌ-ドマウスにおける転移はまれであり, この培養細胞の染色体分析およびアイソザイム分析ではマウス由来の癌と考えられ, また胸腺正常同系マウスへの移植は拒絶されたため, ヌードマウス由来腺癌と考えXK-4細胞と名ずけた.今回, このXK-4細胞の生物学的特性と肺および肝臓への転移能を検討した.

Tumor procoagulant is associated with cancer at advanced stages of malignancy such as infiltration and metastasis. In the present study, we investigated the role of Le y glycolipid in the mechanism of cancer metastasis. Le y glycolipid acts as an important cofactor in the expression of the blood-coagulating activity of cancer cell-derived coagulating activity 1 (CCA-1), which is one of the known tumor procoagulants. Monoclonal antibody (MoAb) FS01, which serves as the Le y -recognizing epitope, inhibits the procoagulant activity of CCA-1 was found to dose-dependently inhibit the procoagulant activity of normal plasma induced by the human lung adenocarcinoma cell line, HAL8, which shows a high level of Le y expression. It did not, however, inhibit the procoagulant activity of the human colon cancer cell line, RPMI4788, which does not express Le y . Administration of FS01 MoAb inhibited lung metastasis of HAL8 cells, but not that of RPMI4788. The absence of antibody-dependent cellular cytotoxicity and complement-mediated cytotoxicity of FS01 MoAb against the HAL8 cell line suggests that the inhibition of HAL8 metastasis by FS01 MoAb derives from the inhibition of blood-coagulating activity of the latter. These findings indicate that Le y glycolipid plays an important role in the mechanism of cancer metastasis via the procoagulant activity of CCA-1. 1.

Galectin-3 is a lactosamine-specific lectin that binds to laminin sugar-sites, and up-regulated expression of galectin-3 in primary colorectal cancer is involved in cancer progression and metastasis. Inhibitory effects of cell adhesion and liver metastasis of adenocarcinoma via portal vein by lectin-binding sugar and anti-galectin-3 antibody was examined to determine the role of galectin-laminin binding in cancer liver metastasis. Highly metastatic adenocarcinoma cell lines XK4-A3 and RPMI4788 were used in in vitro cell attachment and nude mice liver metastatic experiments, and inhibitory effects of anti-galectin-3 antibody or lectin-binding sugars were examined. The in vitro adhesion assay demonstrated that the anti-galectin-3 antibody and alpha-lactose inhibited XK4-A3 and RPMI4788 cell adhesion to laminin in a dose-dependent manner. The liver metastasis of XK4-A3 and RPMI4788 was reduced 50 and 60%, respectively (P

Tumor angiogenesis is an essential step for tumor cell growth, progression and metastasis. Vascular endothelial growth factor (VEGF) is mitogen specific for endothelial cells, and therefore is believed to play a key role in tumor angiogenesis. However, the mechanisms underlying the regulation of VEGF expression remain virtually unknown and the only major regulator of VEGF expression has been reported to be hypoxia. Recently, it was reported that a mutant p53 in#duced the expression of VEGF mRNA, and that wild-type p53 down-regulated endogenous VEGF mRNA levels. In contrast, it has also been reported that mutant ras oncogenes were associated with the marked up-regulation of VEGF in transformed epithelial cells. Based on these results, we performed a retrospective study of the p53 and K-ras genes status and VEGF gene expression in the tumor tissues from 181 patients with non-small cell lung cancer using SSCP, sequencing, RT-PCR and immunohistochemical techniques. Forty-six carcinomas (25.4%) were evaluated as having high VEGF expression, and 135 tumors (74.6%) had low VEGF expression. Of the 181 primary NSCLC studied, 63 carcinomas (34.8%) contained mutations of p53, whereas only 14 carcinomas (7.7%) had mutations of K-ras. There were no significant relationships between VEGF expression and p53 status or each mutant exon of p53. In contrast, a significant difference was found between VEGF expression and K-ras status. Of the 14 tumors with mutant K-ras genes, 7 cases (50.0%) had high VEGF expression whereas only 39 of the 167 tumors with wild-type K-ras (23.4%) had high VEGF expression (p=0.0278). The mean VEGF conservation rate for the 14 tumors with mutant K-ras genes was 0.77+/-0.58 and the rate of the 167 tumors with wild-type K-ras genes was 0.49+/-0.46 (p=0. 0350). Moreover, the overall survival rate of patients with high VEGF expression was lower than patients with low VEGF expression (45.7% vs 60.7%, p=0.0419). On the other hand, there was no significant difference in the overall survival rate between patients with a mutant p53 and those with a wild-type p53; there was also no difference in the overall survival between patients with a mutant K-ras and those with a wild-type K-ras. The Cox regression model analysis indicated that three variables, VEGF status, K-ras status and nodal status, were found to be significant indicators for prognosis (p=0.0236, p=0.0172 and p

Recent investigations have revealed that mutations of the loop-sheet-helix motif of p53 is a significant factor for a poor prognosis in patients with non-small-cell lung cancer (NSCLC). To clarify this mechanism, bcl-2 and bax expression were evaluated in relation to mutations of p53. Tumor tissues of 203 patients with NSCLC were analyzed. Immunohistochemistry was performed to evaluate bcl-2 and bax expression, and polymerase chain reaction single-strand conformation polymorphism following direct sequencing was performed to investigate p53 status. A total of 79 carcinomas were bcl-2 positive, 146 carcinomas were bax positive, and 72 carcinomas had missense mutations of p53. There was no difference in bcl-2 expression in relation to p53 status. On the other hand, tumors with structural mutations of p53 had significantly lower expression of bax than those with wild-type p53 (P = 0.0026). In contrast, tumors with mutations of the loop-sheet-helix motif of p53 had significantly higher expression of bax than those with wild-type p53 (P = 0.0236). The frequency of a bcl-2/bax ratio of ≥1 was significantly lower in tumors with mutations of the loop-sheet-helix motif than that in tumors with wild-type p53 (P = 0.0240). The bcl-2/bax ratio status was a significant factor for a prognosis in patients with NSCLC (P = 0.0083). Mutations of the loop-sheet-helix motif of p53 were correlated with overexpression of bax, while other mutations of p53 were correlated with low levels of bax expression. This variation in pattern of bax expression in relation to mutant p53 might reflect the biological behavior of tumors in patients with bcl-2-positive NSCLC.

It has suggested that hypercoaglability was induced activated factor VII by tissue factor in the estrinsic pathway from malignant tumor. Procoaglant activity may relate with tumor gorwth and development of metastasis. The procoaglant activity of four human cancer cells and of culture supernatant was measured by a one-stage plasma recalcifiation assay. The cell suspension of LK-2, LK-17, LK-52 and HLC-1 was shorter markedly the recalcifiation time of normal human plasma, not of factor VII deficient plasma. The cultre suspension of LK-2 was longer markedly the recalcifiation time of normal human plasma. The culture suspension of LK-17 was shoten markedly the recalcifiation time of normal human plasma, not of factor VII deficient plasma. The culture suspension of LK-52 was shoter the recalficication time of both normal human plasma and factor X, of factor VII deficient plasma. These Beta incident that LK-52 cancer cell line products a directer activater of coagulation facter X in cluture medium.

Association between Ley expression and prognosis of breast cancer was investigated using monoclonal antibody (MoAb) FS01, which recognizes Ley as an epitope, inhibits the procoagulant activity of cancer cell-derived coagulating activity 1 (CCA-1). Expression intensity and procoagulant activity of CCA-1, tissue factor and HLA-DR on breast cancer cell lines were also examined. Immunohistochemical staining of Ley was performed on primary lesions of 223 breast cancer patients who received absolute curative operation. Flow cytometric analysis and clot timer was used to detect expression and activity of each procoagulant on cancer cell lines. The Ley expression was 73.5%, and no significant relation was observed between clinicopathological factors and intensity of Ley expression. The group showing strong Ley positivity had a significantly poorer prognosis than the Ley-negative group in 5-year disease-free survival (p=0.019). Multivariate analysis using the Cox's proportional hazards' regression model showed that Ley expression is an independent prognostic factor (p=0.018), following tumor size and lymph node metastasis. Ley expression on cancer cell surface is higher than tissue factor and HLA-DR. FS01 and anti-tissue factor MoAb inhibited the coagulating activity of tissue factor-expressing lines, but no cells were inhibited by staphylococcal enterotoxin A, which is known to inhibit the coagulating activity of HLA-DR. CCA-1 and tissue factor plays a important role in the blood coagulating activity of breast cancer cell lines. Breast cancer patients are thought to have a poor prognosis because Ley expression on the surface of the cancer cell induces blood coagulation via CCA-1.

As part of our evaluation of MRP-1/CD9 and KAI1/CD82 as prognostic predictors among patients with cancer, we have extended our studies to solid tumors of a variety of anatomical sites. Normal tissues were included for comparison. Immunohistochemical techniques were used throughout. Our results indicate that MRP-1/CD9 was strongly expressed by many normal tissues, including the epithelium of the gastrointestinal tract, alveolar epithelium of the lung, urothelium and smooth muscle. Expression was weak in the pituitary gland, spleen and hepatocytes, and absent in testes and spinal cord. KAI1/CD82 was also expressed by many normal tissues, but was absent in some MRP-1/CD9-positive tissues (e.g., smooth muscle, adrenal cortex, urothelium, myelin of peripheral nerves, epithelium of amnion). On the other hand, KAI1/CD82 was strongly expressed in spinal cord gray matter, which was MRP-1/CD9-negative. Expression of these glycoproteins was detected in almost all types of tumors examined. In certain cancers, MRP-1/CD9 and KAI1/CD82 positivity was inversely related to lymph node involvement. Whereas lymph node metastases were present in 22.2% of lung cancer patients whose tumors were MRP-1/CD9 and KAI1/CD82-positive, 65.5% of patients with MRP-1/CD9 and KAI1/CD82-reduced/negative tumors had lymph node metastases. A similar inverse relationship was seen in colon cancer and breast cancer patients with respect to MRP-1/CD9 expression. The present data, together with our previous results suggest that evaluating the MRP1/CD9 and KAI1/CD82 status of cancers of the lung, breast and colon may provide useful information on the metastatic potential of the tumors.

A protein which enhanced urokinase-type plasminogen activator (u-PA) activity was purified from the extracts of extracellular matrix of highly metastatic cell line HAL-8 derived from human lung adenocarcinoma. The protein showed a single band with molecular weight of 65 kDa after the purification by Sephadex G-150 and diethylaminoethyl-cellulose followed by reversed phase separation in a high performance liquid chromatography system. The purified protein in the immobilized conditions enhanced u-PA activity in both plasminogen activation and S-2444 amidolysis by 4.6- and 2.8-fold increases in the second order rate constants (Kcat/K(m)), respectively. This protein was related to neither plasminogen nor single-chain u-PA by the immunological studies and with respect to retention time on reversed phase analysis. These results suggest that the purified material acts as an enhancer of u-PA in extracellular matrix of the cancer cells, inducing an effective tissue destruction and cell invasion and possessing a highly metastatic potential.

PURPOSE: The usefulness of postoperative carcinoembryonic antigen (CEA) monitoring and improvements in imaging techniques have renewed enthusiasm for second-look operations (SLO) as the most effective treatment for recurrent colorectal cancer by reresection following early detection. The aim of our study is to evaluate the role of CEA and imaging techniques-directed SLO. METHODS: Seven hundred fifty-six patients with Dukes Stages B and C, who had undergone curative resection, were monitored postoperatively using CEA and imaging techniques. An SLO was performed on any potentially resectable recurrence, and in addition, an SLO was done when a persistently rising CEA value was detected. RESULTS: Recurrence developed in 18.8 percent (142/756) of patients, and 90.8 percent (129/ 142) of the recurrences were detected within the first three years following curative resection. When comparing carcinomas of the colon with that of the rectum, the former were associated with significantly more hepatic and intra-abdominal recurrences, whereas the latter had significantly more locoregional and pulmonary recurrences. Seventy-two patients underwent SLO. Of these patients, 54.2 percent (39/72) had all of their disease resected, and 1.4 percent (1/72) had no detectable disease at the SLO. Among the 142 patients with recurrence, 71 (50 percent) patients underwent SLO. The resectable group at SLO carried a significantly better survival than the unresectable recurrence group (41.3vs.5.2 percent;P

PURPOSE: The aim of this study was to clarify the distribution of lymph node metastasis in colorectal cancer. We also examined the relationship between the primary tumor (T) and the regional node (N) categories of the TNM (primary tumor, regional nodes, metastasis) classification. METHOD: Lymph nodes of surgical specimens in 311 consecutive patients with colorectal cancer were studied using the modified clearing method. RESULTS: Lymph node metastasis was seen in 59.2 percent of the total cases. The upward metastasis rate was 30.7 percent. In the longitudinal spread, most of the lymph node metastasis was seen within 10 cm. On the oral side in rectal cancer, there was no metastasis beyond 4 cm. The lateral metastasis rate in rectal cancer was 8.8 percent and in the lower rectum, the rate of cancer within 6 cm from the anal verge or beyond pT3 was much higher. CONCLUSION: In the TNM classification, there was no significant difference between colon and rectal cancer except pT1 with rectal cancer. In the lower rectal cancer within 6 cm from the anal verge or beyond pT3, there is a high risk of lateral metastasis, and lateral lymph node dissection or radiation therapy should be performed.

In anterior resection with anastomosis using the double-staple technique for low-lying rectal cancer in male patients, the approach to the anal canal with a stapling instrument via the abdominal area is limited by the narrow pelvis. The stapling and transection of the anal canal via the posterior transsacral approach prior to performing an anterior resection thus enables the lower rectum and anal canal to be visualized, so that the anal canal can be accurately stapled and transected even in male patients with a narrow pelvis.

胃癌患者の腹腔滲出細胞を採取してその抗腫瘍活性の測定とOK432術前投与によるその変動効果を検討した.平均回収細胞数は (2.2±0.4) ×107個で各臨床病期間で有意差を認めなかった.抗腫瘍活性の測定として, (1) lymphokine-activated killer (LAK) 誘導能, (2) 腹腔マクロファージの腫瘍増殖抑制能を検討したがLAK誘導能ではいずれの症例からも有意な活性は認められず, またOK432術前投与にても効果は認めなかった.腫瘍増殖抑制能ではStage II, III症例で比較的高い活性を認め (Stage I 16.3±6.0%, Stage II 45.0±4.4%, Stage III 23.0±9.0%, Stage IV 12.2±10.2%) またOK432投与にてその活性増強を認めたものの総体的にその抗腫瘍活性は高くなく, 腹腔滲出細胞の局所防禦に占める役割は大きいものではないと考えられた.

It has been suggested in experimental animal cancer system that platelet aggregating potential of cancer cell and platelet aggregating material (PAM) is one of the most important factor in cancer metastasis. We established a human lung cancer cell line (KUM·LK-2) which produce spontaneous lung metastasis in nude mouse. PAM was extracted from KUM·LK-2 cell and PAM owned aggregating potential in human heparinized-PRP. PAM was differ from well known aggregating materials like ADP, collagen and others. PAM didn't contain fibrinogen and sialic acid, and was not concerned witht hromboxan composition. PAM was considered high molecular weight protein itself containing phospholipid and aggregating potential was concerned with ATP composition of platelet. This was first confirmation of platelet aggregating potential in human cancer cell line which own metastatic potential. Platelet aggregating potential of KUM·LK-2 cell may play a important role in blood borne metastasis.

胃癌切除症例130例にクリアリング法を用いてリンパ節のsinus histiocytosisについて検索し, 癌転移様式との関係を検討した. クリアリング法による検索リンパ節数は7,465個で転移リンパ節数743個, 転移度10.0%, 1症例当りの検索リンパ節数は57.4個で従来の触診法の25.5個に比べ倍以上に増加した. sinus histiocytosisはリンパ節が大きくなるほど, また, 第1群から第3群リンパ節へと高次になるに従い. 高くなった. 年齢. 癌占居部位・肉眼型・組織型には影響されず, 癌のstageと関係したが, 癌の深達度よりもリンパ節転移により深い関係が認められた. このことはsinus histiocytosisの局所における防御反応としての意義を示唆した.

癌の転移に関与する因子には癌細胞側因子, 宿主側因子, 両者の相互作用などがあるが, 今回, 肺癌培養細胞3株を用い, 癌細胞側因子 (血小板凝集育旨, plasminogen activator産生能) とnude mice皮下移植での肺転移形成能との相関について検討した.血小板凝集能, urokinase産生能は3株共に見られたが, 自然肺転移を生じるKUM.LK-2細胞はtissue plasminogen activatorをも産生しており, この物質の産生能が肺転移形成に関与していると推測された.

The usefulness of a new tumor marker, KM01, the monoclonal antibody of which, for established cells of human colonic carcinoma, detects the presence of tumor, was determined using both clinical procedures and an experimental tumor line, and was compared with those of CA19-9, CEA and IAP. The measurement of KM01-RPHA was found to be convenient, showing positive rates of 80.0% and 44.0% in cases of pancreatic carcinoma and gastric cancer, respectively, and furthermore showing a high positive rate of 80.0% in cases of relapsed gastric and colonic carcinomas. Consequently KM01 seems to be useful as a tumor marker. The results of measurements with KM01 and CA19-9 were consistent and the correlation between antibody values of KM-01-RPHA and measurements with CA19-9 was high. The false-positive rate for KM01 was higher than that for CA19-9, suggesting that KM01-RPHA contains antigenic determinants similar to those of CA19-9, facilitating measurement over a wider range than in the case of CA19-9. The antibody value of KM01-RPHA was correlated with CEA but not with IAP. In culutured sera of three kinds of established cancer cells, release of KM01 was found. Among these KUM MK-2 was transplanted into nude mice, and release of KM01 into the sera of the nude mice was initially recognized.

We reported previously the establishment and platelet aggregating activity of human lung adenocarcinoma cell line KUM·LK-2 which produce spontaneous metastasis in nude mouse. Platelet aggregating potential in vivo and vitro, and metastatic potential of KUM·LK-2 were performed under using anti-platelet agent DN 9693 which owned phosphodiesterase blocking activity and not owned anti-cancer activity. DN 9693 blocked platelet aggregating potential and fatal acute thromboembolism of nude mouse induced by KUM·LK-2 cell. KUM·LK-2 metastases were produced subcutaneous implantation group only. But in the case of naturall killer remission using anti-asialo GM1, KUM·LK-2 produce metastases intravenous injection group too. DN 9693 blocked the metastasis of KUM·LK-2 in the case of natural killer reduced nude mouse completely. From these results DN 9693 blocked platelet aggregating activity of KUM·LK-2 cell in vivo and vitro, and blocked metastasis. It was concluded that platelet aggregating activity of KUM·LK-2 cell is necessery to produce metastasis. It was first experiment of human cancer metastasis blocking using anti-platelet agent and proof that anti-platelet agent may usefull for anti-metastasis of human cancer.

It has been reported in animal experimental system that Platelet Aggregating Material (PAM) of cancer cell play important role in cancer metastasis. Many human cancer cell lines has been also studied the platelet aggregation activity of PAM. But correlation between the platelet aggregation activity and metastatic potential of human cancer cells were usually unkncwn. We established a human lung adenocarcinoma cell line KUM-LK-2 which produce spontaneous lung metastasis when cells implanted into subcutaneous of nude mice. PAM was extracted from KUM-LK-2 cells following the method of D.Mbhanty and P.Hilgard. Platelet aggregation study of PAM was performed by human Heparinized platelet rich plasma useing NIKO Hematracer PACT2D. Character of PAM were examined by physical and chemical treatment. KUM-LK-2 PAM shew 80% of platelet aggregation in maximum after 150 sec lag time, and aggregation was not found by Citrated PRP.It is concluded that PAM is high moleculer protein and contain Phospholipid and aggregation activity is concerning with ATP composition of platelet. PAM dose not contain Fibrinogen and Sialic acid, and not concern with Throrriboxan composition. This study is first case of platelet aggregation activity of PAM extracted human canoer cells which contain metastatic potential.