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Background: The most widely used noninvasive screening tests for colorectal cancer are fecal occult blood tests. Stool DNA test was developed in recent years. However, direct comparative analyses of these tests within the same population are still sparse. Methods: A total of 2,842 participants who visited outpatient clinics or cancer screening centers were enrolled. Stool DNA test-I (KRAS, BMP3, NDRG4, and hemoglobin immunochemical tests), stool DNA test-II (SDC2 and SFRP2 tests), and fecal immunochemical test (FIT) alone were performed and colonoscopy was used as the gold standard among 2,240 participants. Forty-two and 302 participants had colorectal cancer and advanced adenomas (AA), respectively. Results: The sensitivity for colorectal cancer of stool DNA test-I, -II, and FIT was 90.5%, 92.9%, and 81.0%, respectively. The sensitivity for advanced neoplasm (AN; colorectal cancer plus AA) of stool DNA test-I, -II, and FIT was 34.9%, 42.2%, and 25.9%, respectively. The specificity of stool DNA test-I, -II, and FIT was 91.4%, 93.3%, and 96.8%, respectively, among those with negative results on colonoscopy. When the specificity of FIT was adjusted to match that of stool DNA tests by changing the threshold, no significant difference was seen in the sensitivities among the three tests for detecting colorectal cancer. For AN, the sensitivity of FIT was higher than DNA test-I and similar to DNA test-II under the same specificities. Conclusions: There was no significant advantage of the two stool DNA tests compared with FIT in detecting colorectal cancer or AN in this study. Impact: Our findings do not support extensive use of stool DNA tests instead of FIT.

The Asia-Pacific Colorectal Screening (APCS) scoring system was developed to stratify the risk of colorectal advanced neoplasm (AN). We aimed to evaluate the performance of the APCS score combined with a stool DNA test used for colorectal cancer screening.A total of 2842 subjects who visited outpatient clinics or cancer screening centers were enrolled. Age, sex, smoking status, and family history were recorded and APCS scores were calculated in 2439 participants. A stool DNA test (SDC2 and SFRP2 tests) and fecal immunochemical test (FIT) were performed and colonoscopy was used as the gold standard among 2240 subjects who completed all study procedures. We used a threshold of 4.4 μg/g for the FIT, in addition to the manufacturer's recommended threshold of 20 μg/g to match the specificity of a stool DNA test.Based on the APCS score, 38.8% (946 of 2439) of the subjects were categorized as high risk, and they had a 1.8-fold increase in risk for AN (95% CI, 1.4-2.3) compared with low and moderate risk. The APCS combined with the stool DNA test detected 95.2% of invasive cancers (40 of 42) and 73.5% of ANs (253 of 344), while the colonoscopy workload was only 47.1% (1056 of 2240). The sensitivity for AN of APCS combined with stool DNA test was significantly higher than that of APCS combined with FIT (73.5% vs 62.8% with FIT cut-off value of 20 μg/g, and 73.5% vs 68.0% with FIT cut-off value of 4.4 μg/g; both P.01).The APCS score combined with a stool DNA test significantly improved the detection of advanced colorectal advanced neoplasms, while limiting colonoscopy resource utilization (Chictr.org.cn, ChiCTR-DDD-17011169).

Limited literature exists on the characteristics of early gastric signet ring cell carcinoma (GSRCC) within 20 mm. This study aimed to explore this type of cancer from several aspects, to provide guidance for early detection and intervention of GSRCC.We retrospectively collected data from 24 patients diagnosed with early GSRCC ≤20 mm in Beijing Friendship Hospital from 2016 to 2021. According to tumor size, those lesions were divided into three groups: diminutive group (1-5 mm,Treatment strategies for lesions differed according to the size (The location of early signet ring cell carcinoma is not specific, and the diminutive lesions are often flat. Abnormal IP may be the early endoscopic feature of early GSRCC.

Introduction: Asia-Pacific Colorectal Screening (APCS) score has been implemented for colorectal cancer screening in asymptomatic cohort in many regions. However, no study has validated its efficiency in Asian outpatients with mild-self-limited gastrointestinal symptoms yet. The purpose of this study was to validate its efficiency in asymptomatic subjects and outpatients in Ningxia. Methods: The records of 329 asymptomatic participants and 300 outpatients were collected and analyzed from database in the General Hospital of Ningxia Medical University from September 2017 to April 2018. These 2 main groups were divided into 3 tiers based on the scores calculated by the category of APCS score. The detection rates of advanced colorectal neoplasia (ACRN) were further compared according to histopathological classifications of tissues acquired during colonoscopy. Results: Among the 329 participants screened in the asymptomatic cohort, 78 subjects (23.7%) were in the low-risk (LR) tier, 187 subjects (56.8%) in the moderate-risk (MR) tier, and 64 subjects (19.5%) in the high-risk (HR) tier. The percentage of ACRN in the LR, MR, and HR groups was 1.3, 8.6, and 20.3%, respectively. In the 300 outpatient cohorts, 78 patients (26%) were in the LR tier, 140 patients (46.7%) in the MR tier, and 82 patients (27.3%) in the HR tier. The detection rates of ACRN in the LR, MR, and HR groups were 0, 10, and 39%, respectively. Conclusion: APCS score is an effective method for ACRN screening in asymptomatic and also the outpatient subjects. Individuals with HR scores should be given priority for colonoscopy.

Background Colorectal endoscopic submucosal dissection (ESD) is a challenging operation because it is often difficult to attain good visualization of the submucosal layer. Lots of traction methods have been developed to facilitate submucosal dissection, but still not widely applied. Therefore, we designed a new traction device "traction ring". This pilot study is to evaluate if traction ring is feasible and safe for colorectal ESD. Methods We retrospectively analyzed 20 patients with colorectal lesions who had undergone traction ring assisted ESD. The main outcome was the rate of en bloc resection, R0 resection rate, procedure time, resection time, intraoperative and postoperative complications. Results The median procedure time was 74.5 min (range 35–269 min.). The median resection time was 55 min (range 25–209 min.). The application of the traction system accounting for only 2.7% of the whole procedure time. Rate of en bloc resection was 95% (19/20). Rate of R0 resections was 90% (18/20). All traction rings were successfully settled and retrieved. No serious excessive intraoperative bleeding was observed. One patient was found perforation after treatment, no further intervention was required. No delayed complications were observed within one month after the operation. Conclusions This retrospective study demonstrated that the new countertraction method (traction ring) may become a safe, effective and simple way to assist ESD for colorectal lesions.

Objectives To investigate the efficacy and safety of premedication with simethicone/pronase during esophagogastroduodenoscopy (EGD) with sedation. Methods 610 patients were randomly allocated to two groups based on the type of premedication given. The premedication used in the control group was 10ml of lidocaine hydrochloride mucilage (LHM, N=314) and the premedication used in the intervention group was 80mL of simethicone/pronase solution plus 10ml of lidocaine hydrochloride mucilage (SP/LHM, N=296). The EGD was performed under sedation. The visibility scores, number of mucosal areas that need to be cleansed, water consumption for cleansing, time taken for the examination, diminutive lesion, pathological diagnosis, patients’ gag reflex and the oxygenation (pulse oximetry) were recorded. Results The SP/LHM has significantly lower total visibility score than the LHM (7.978±1.526 vs 6.348±1.097, p

Background Detecting gastric cancer in the early stage is the key factor in reducing cancer mortality. Endoscopists can now detect early cancer with the aid of advanced imaging in endoscopy. A major technical obstacle in achieving a good quality of endoscopic examination of the gastric mucosa is the presence of foam and mucus over the mucosal surface. Pronase has become a common anti-mucus agent used for improving visibility during upper gastrointestinal endoscopy examination, especially in eastern Asia. While N-acetylcysteine (NAC) can serve as an alternative mucolytic agent more economically. The aim of this study is to investigate the efficacy and cost-benefit of premedication with NAC during esophagogastroduodenoscopy (EGD). Methods We enrolled 1,200 patients and randomly allocated them into 4 groups with different types of premedication. Premedication used in regular group (RG) was 10 mL lidocaine hydrochloride mucilage (LHM) and in pronase+simethicone group (P+S) was 60 mL pronase/simethicone solution plus 10 mL LHM. Meanwhile, 60 mL NAC600 mg/simethicone solution plus 10 mL LHM and 60 mL NAC1200 mg/simethicone solution plus 10 mL LHM were given to the patients in NAC600+simethicone group (NAC600+S) and NAC1200+simethicone group (NAC1200+S), respectively. We recorded visibility scores, times of cleaning, time taken for examination, diminutive lesions and pathological diagnosis. Cost-benefit of premedication was analyzed. Results The visibility of NAC600+S and NAC1200+S are significantly better than RG, although P+S exhibited the lowest total visibility score and shortest procedure duration (figure 1). Times of cleaning was remarkably decreased in NAC600+S and NAC1200+S compared to RG, but was inferior to P+S. There is no significant difference in detection rate of diminutive lesions and pathological diagnosis. Cost-benefit analysis has shown that net medical yield of NAC/simethicone is higher than that of pronase/simethicone. Conclusions NAC can not only facilitate the visibility during EGD, but be more cost-beneficial. It should be recommended as a good alternative premedication to pronase during EGD, especially in developing countries and regions.

Sporadic tumors of the duodenal papilla are rare, with a prevalence of 0.04% to 0.12% in autopsies [1], most of which are adenomas histologically [2].

Background Calcyclin Binding Protein/(Siah-1 interacting protein) (CacyBP/SIP) acts as an oncogene in colorectal cancer. The nuclear accumulation of CacyBP/SIP has been linked to the proliferation of cancer cells. It has been reported that intracellular Ca2+ induces the nuclear translocation of CacyBP/SIP. However, the molecular mechanism of CacyBP/SIP nuclear translocation has yet to be elucidated. The purpose of this study was to test whether the Ca2+-dependent binding partner S100 protein is involved in CacyBP/SIP nuclear translocation in colon cancer SW480 cells. Methods The subcellular localization of endogenous CacyBP/SIP was observed following the stimulation of ionomycin or BAPTA/AM by immunofluorescence staining in SW480 cells. S100A6 small interfering RNAs (siRNA) were transfected into SW480 cells. Immunoprecipitation assays detected whether S100 protein is relevant to the nuclear translocation of CacyBP/SIP in response to changes in [Ca2+]i. Results We observed that endogenous CacyBP/SIP is translocated from the cytosol to the nucleus following the elevation of [Ca2+]i by ionomycin in SW480 cells. Co-immunoprecipitation experiments showed that the interaction between S100A6 and CacyBP/SIP was increased simultaneously with elevated Ca2+. Knockdown of S100A6 abolished the Ca2+ effect on the subcellular translocation of CacyBP/SIP. Conclusion Thus, we demonstrated that S100A6 is required for the Ca2+-dependent nuclear translocation of CacyBP/SIP in colon cancer SW480 cells.

This study retrospectively compared the safety and efficacy of two endoscopic techniques for treating newly diagnosed achalasia, pneumatic dilation (PD), and peroral endoscopic myotomy (POEM). Demographics, clinical and manometric data, and outcomes were collected from the medical records of patients who received POEM or PD as the primary therapy for achalasia at our hospital from January 2012 to August 2015. Of 72 patients, 32 and 40 received POEM and PD, respectively. The two groups had similar preoperative features. On short-term follow-up, improvements in high-resolution esophageal manometry and barium esophagogram parameters were similar. For PD, the success rates at 3, 6, 12, 24, and 36 months were 95, 88, 75, 72, and 60%, respectively. For POEM, these were 96, 96, 96, 93, and 93% (P = 0.013, log-rank test). On subgroup analysis, the success rate was higher with POEM than that with PD in all 3 manometric subtypes, but only that of type III was statistically significant. POEM required significantly longer operative time and hospitalization than did PD (P

Introduction : The incidence of ulcerative colitis has increased tremendously over the past few decades, and caused much more attention in China. Several studies have revealed that both environmental and genetic factors play crucial roles in the formation and development of this disease. Among the environmental factors, the smoking condition is highly significant. Although current smoking has been associated with a protective effect on both the development and progression of ulcerative colitis (UC), the clinical characteristics of UC patients with current smoking in China is still unclear. Aim : To clarify the relationship between current smoking and the UC in the northwest of China, patients diagnosed with ulcerative colitis in the General Hospital of Ningxia Medical University from 2003.03 to 2012.09 were assessed. Material and methods : Data were collected about clinical features, cigarette smoking conditions, disease location, and therapeutic requirements. A total of 421 patients were included in this study, at diagnosis, 341 (81%) patients were non-smokers, and the current smokers amounted to 80 (19%). Results : As for disease location, there were no differences between the current smokers and non-smokers regarding proctitis and extensive colitis, but the current smokers had less left-sided colitis than non-smokers (p = 0.049). Based on the Modified Mayo score, heavier smokers had lower scores than other smokers and non-smokers. For the therapeutic requirements, current smokers needed fewer corticosteroids than non-smokers (p = 0.048). Conclusions : We found that current smoking is closely associated with a protective factor for UC.

Cyclooxygenase-2 (COX-2), the key enzyme in prostaglandin synthesis, is often over-expressed in human gastric cancer. Recently, 15-hydroxyprostaglandin dehydrogenase [NAD+] (15-PGDH), the key enzyme in prostaglandin degradation, was found to be down-regulated in human gastric cancer tissues, but little is known about its role in gastric tumorigenesis. In this study, expression plasmids containing 15-PGDH siRNA were constructed and transfected into the gastric cancer cell line MKN45, which expresses endogenous 15-PGDH at a high level. The 15-PGDH gene was also transfected into the gastric cancer cell line SGC7901, which expresses endogenous 15-PGDH at a low level. When compared with the empty vector transfectant, MKN45 cells stably transfected with the 15-PGDH siRNA plasmid had a significantly increased proliferation rate. In contrast, SGC7901 cells stably transfected with the 15-PGDH cDNA had a significantly decreased growth rate. Furthermore, increased expression of 15-PGDH suppressed clone formation of gastric cancer cells in plate and soft agar colony formation assays in vitro and suppressed tumor formation in athymic nude mice in vivo. Stable silencing of 15-PGDH in gastric cancer cells also enhanced cell cycle entry in vitro. These results demonstrate for the first time that 15-PGDH acts as a tumor suppressor in human gastric cancer and provide further validation for 15-PGDH as a potential therapeutic target for human gastric cancer.

Background and aims HER2 , an oncogene, has been found to be over-expressed in 10–40% of human gastric carcinomas. The aims of this study were to investigate if a fusion protein consisting of anti-HER2 sFv and constitutively active caspase-3 was capable of inducing apoptosis in HER2-expressing human gastric cancer cells and blocking the growth of human gastric cancer xenografts in nude mice. Methods NIH3T3 cells stably transduced with the pcDNA3.1-HER-PE-CP3 recombinant plasmid containing a secretion signal, a single-chain anti-HER2 monoclonal antibody fragment, a Pseudomonas exotoxin A translocation domain and a constitutively active caspase-3 molecule were used to induce apoptosis in human gastric cancer cells both in vitro and in vivo. Immunofluorescence staining and western blotting were used to examine the expression of the recombinant protein HER-PE-CP3. Apoptosis was determined by flow cytometry and TUNEL assay. Results Co-cultivation of HER-PE-CP3/ NIH3T3 with human gastric cancer cells led to internalisation of HER-PE-CP3 and apoptosis in HER2-expressing human gastric cancer cells but not in HER2-negative cancer cells. Inoculation of HER-PE-CP3/NIH3T3 in nude mice resulted in potent inhibition of human gastric cancer xenografts and much prolonged survival time of the tumour-bearing mice compared with the control. Significantly more apoptotic cells were detected in xenografts in mice receiving HER-PE-CP3/NIH3T3 than in control mice. Conclusions The HER-PE-CP3 chimeric molecule could induce selective apoptosis and potent growth inhibition of HER2-positive human gastric cancer cells and might represent a novel HER2-directed treatment option for human gastric cancer.

Calcyclin-binding protein or Siah-1-interacting protein (CacyBP/SIP), a component of the ubiquitin-mediated proteolysis, could bind SKP1-CUL1-F box protein complex and participate in beta-catenin degradation, which was found to be related to the malignant phenotypes of gastric cancer and renal cancer. However, the role of CacyBP/SIP in pancreatic cancer progression still remains unclear. Therefore, the aim of the present study was to investigate the expression and clinical significance of CacyBP/SIP in pancreatic cancer.Immunohistochemistry was carried out on paraffin-embedded sections of pancreatic cancer and normal pancreatic tissues. In addition, Western blot and semiquantitative RT-PCR were carried out to analyze mRNA and protein expression of CacyBP/SIP in 8 pairs of freshly resected pancreatic cancer and their adjacent nontumorous tissue.CacyBP/SIP expression was significantly increased in pancreatic cancer tissue (28/68 or 41.2%) and correlated with differentiation degree, higher TNM (tumor, node, metastasis) stage and distance metastasis. Also, mRNA and protein expression of CacyBP/SIP were found to be at higher levels in almost all cancer tissues compared to adjacent tissues.CacyBP/SIP protein might play an important role in the process of pancreatic carcinogenesis and high-level CacyBP/SIP expression might be related to the malignant potential of pancreatic cancer.

Multidrug resistance (MDR) is a major impediment to the effective chemotherapy of many human malignancies. Although much effort has been devoted to develop new drugs for overcoming MDR, until now, still no useful method of reversing MDR, suitable for clinical use, has emerged from this large quantity of work. Some researchers have reported that proteasome inhibitors could induce apoptosis in a variety of cancer cells. In the present study, we found that, in vincristine-resistant human gastric cancer cell line SGC7901/VCR, proteasome inhibitor MG132 was an effective inducer of apoptosis, and also had the capacity of downregulating the expression of anti-apoptotic Bcl-2 and MDR1 (P-gp), by which MG132 resensitized tumor cells to the apoptosis induced by anticancer drugs. Data presented by drug sensitivity assay further demonstrated that MG132 could reverse the resistant phenotype of gastric cancer cells effectively through both enhancing drug-induced apoptosis and inhibiting P-gp. The further study of the effectiveness and safety of proteasome inhibitor in vivo may be helpful for developing a new possible strategy to treat gastric cancer MDR.

Osteopontin (OPN), cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) are overexpressed in various experimental models of malignancy. However, the correlation and role of the three molecules in gastric cancer is unclear. In the present study, we found that OPN, COX-2 and VEGF were overexpressed in 53 cancerous tissues with gastric cancer compared with 40 normal mucosa tissues by immunohistochemistry method. Moreover, the results indicated co-expression of OPN, COX-2, and VEGF in gastric cancer. Levels of OPN, COX-2, and VEGF were all significantly correlated with TNM stage, lymph node metastasis and distant metastasis (P0.05), while not related to prognosis of patients. In addition, individual levels of OPN, COX-2, and VEGF were all significantly correlated with microvessel density (MVD), valued by CD34 staining directly with r-values of 0.416, 0.400, and 0.566, respectively (P0.01). Both OPN and COX-2 levels showed a positive correlation with VEGF (P0.05). Meanwhile, expression of COX-2 is in relation to OPN (P0.01). Overall, survival for patients with high MVD was significantly lower than for patients with low MVD (P0.05). Our findings indicate that OPN, COX-2, and VEGF synergically promote angiogenesis and metastasis in gastric cancer. It may be an important and useful strategy to target these molecules for prevention and therapy of tumor.

We have investigated the roles of dopamine and cAMP-regulated phosphoprotein (DARPP-32) in the multidrug resistance (MDR) of gastric cancer cells and the possible underlying mechanisms. The up-regulation of DARPP-32 was found to significantly enhance the sensitivity of cells of human adriamycin (ADR)-resistant gastric adenocarcinoma cell line SGC7901/ADR to vincristine, ADR, 5-fludrouracil and cisplatin. The results of an in vivo drug sensitivity assay confirmed that DARPP-32 may play a specific role in the MDR of gastric cancer. DARPP-32 significantly down-regulated the expression of P-glycoprotein and zinc ribbon domain-containing 1 (ZNRD1), but did not alter the expression of MDR-associated protein or glutathione-S-transferase. The up-regulation of ZNRD1 significantly inhibited the drug sensitivity of gastric cancer cells over-expressing DARPP-32, indicating that ZNRD1 may be important in the DARPP-32-mediated MDR of gastric cancer. DARPP-32 was also able to significantly decrease the anti-apoptotic activity of SGC7901/ADR cells. Further study of the biological functions of DARPP-32 may be helpful for understanding the mechanisms of MDR of gastric cancer cells and developing possible strategies to treat gastric cancer.

The p75 neurotrophin receptor (p75NTR) is a focus for study at present. However, its function in gastric cancer was not elucidated. Here, we investigated its relation with metastasis of gastric cancer. By immunohistochemistry, we found that the positive rate of p75NTR expression in metastatic gastric cancer was 15.09% (16 of 106), which was lower compared with nonmetastatic gastric cancer (64.15%; 68 of 106). The average staining score in nonmetastatic gastric cancer was significantly higher than in metastatic gastric cancer (1.21 ± 0.35 versus 0.23 ± 0.18; P < 0.01). p75NTR protein level was also lowly expressed in the highly liver-metastatic gastric cancer cell line XGC9811-L compared with other gastric cancer cell lines by Western blotting. It could also significantly inhibit the in vitro adhesive, invasive, and migratory and in vivo metastatic abilities of gastric cancer cell lines SGC7901 and MKN45 by reducing urokinase-type plasminogen activator (uPA) and matrix metalloproteinase (MMP)-9 proteins and by increasing tissue inhibitor of matrix metalloproteinase (TIMP)-1 protein. Further studies showed that p75NTR could suppress the nuclear factor-κB (NF-κB) signal. SN50, a specific inhibitor of NF-κB, which could inhibit in vitro invasive and migratory abilities of gastric cancer cells, reduced expression of uPA and MMP9 proteins and increased expression of TIMP1 protein. Taken together, p75NTR had the function of inhibiting the invasive and metastatic abilities of gastric cancer cells, which was mediated, at least partially, by down-regulation of uPA and MMP9 proteins and up-regulation of TIMP1 protein via the NF-κB signal transduction pathway. Our studies suggested that p75NTR may be used as a new potential therapeutic target in metastatic gastric cancer. (Mol Cancer Res 2007;5(5):423–30)

Angiogenesis plays a crucial role in tumor development and growth. The present study was carried out to investigate the potential involvement of the cyclooxygenase-2 (Cox-2) pathway in the regulation of angiogenesis in hepatocellular carcinoma (HCC). We inhibited Cox-2 expression in HCC cell line HuH-7 by selective Cox-2 inhibitor (SC-58635) or Cox-2 siRNA. Conditioned media (CMs) from HuH-7 cells were used in angiogenic assays in vitro and in vivo. Compared with CMs from untreated and negative siRNA treated HuH-7 cells, CMs from SC-58635 and Cox-2 siRNA treated HuH-7 dramatically suppressed the proliferation, migration, and differentiation of human umbilical vein endothelial cells (HUVECs) in vitro and neovascularization in vivo. These inhibitory effects could be partially reversed by the addition of exogenous PGE2 to CMs. Furthermore, Cox-2 inhibition by SC-58635 resulted in PGE2 reduction accompanied by the down-regulation of four PGE2 receptor (EP receptor) subtypes. Treatment with SC-58635 led to the down-expression of proangiogenic factors such as VEGF, HGF, FGF2, ANGPT1 and ANGPT2 in HCC. An approximately 78% reduction of VEGF level has been found in the CM from SC-58635 treated HuH-7. Our results suggest an involvement of Cox-2 in the control of HCC-associated angiogenesis. PGE2 as a vital angiogenic factor may act directly on endothelial cells to promote HuH-7-stimulated angiogenic process. Moreover, Cox-2/PGE2/EP/VEGF pathway possibly also contributes to tumor angiogenesis in HCC. This study provides the rationale for clinical studies of Cox-2 inhibitors on the treatment or chemoprevention of HCC.

Peritoneal dissemination in gastric cancer is the most frequent cause of the noncurative resection and recurrence after curative resection. We, therefore, evaluated the feasibility of a peptide, which was obtained by screening a random phage display library, in the treatment of peritoneal metastases of gastric cancer. In this study, a novel cell line, GC9811-P, with a high potential peritoneal metastasis of gastric cancer derived from its parental cell line, GC9811, was established. Using a phage display library, we isolated a specific peptide that selectively bound to GC9811-P cells rather than its parental GC9811cells. The isolated phage-displaying peptide, SMSIASPYIALE (named peptide PIII), was obtained after four rounds of selection, showing a tendency to preferentially bind to GC9811-P cells compared with a panel of other gastric cancer cell lines, and preferentially accumulate in peritoneal metastasis tumor tissue in comparison with control organs, peritoneum, liver, pancreas, spleen, lung, and kidney. Further study showed that synthetic peptide PIII could significantly inhibit adhesive and invasional ability of GC9811-P cells and could effectively block the corresponding phage binding to the GC9811-P cells, whereas, exposure of the cells to various concentrations of peptide PIII showed no obvious cell growth inhibition. Furthermore, a highly reproducible animal experimental model of gastric cancer with peritoneal dissemination was established in nude mice by injecting a suspension of the cell line into the gastric wall of nude mice. Animals intraperitoneally treated with peptide PIII in this model or another animal model of gastric cancer with peritoneal dissemination established using MKN45 cells showed suppressed tumor metastasis to peritoneum and significantly prolonged survival. In conclusion, the selected peptide PIII was a biologically active peptide and could effectively inhibit peritoneal dissemination of gastric cancer.

Tumor growth, progression and metastasis are critically dependent on blood supply, which has received increased attention as a potential target of new anticancer therapies. Antiangiogenic therapy to limit and even reverse the growth of tumors is under investigation and showing promise. Moreover, tumor vascular express specific surface proteins (“vascular zip codes”), not present in resting blood vessels of normal tissues, are suitable for targeting purposes. The specific “vascular zip codes” can be identified by screening phage-displayed peptide library in vivo. This technology is simple but powerful, providing the advantages of selectivity of action plus improved accessibility and efficacy. To date, a variety of tumor-homing peptides have been isolated in this method, and most of the peptides have been used for targeting devices to concentrate drugs and other therapeutic materials to tumors. Such a targeting strategy can decrease toxicity and increase the therapeutic efficacy of the drug.

Anti-calcyclin binding protein (CacyBP) monoclonal antibodies (MAb) were produced using an in vitro immunization method. BALB/c mouse splenocytes were immunized with purified 6 x His-CacyBP fusion protein and fused with myeloma cells using polyethylene glycol (PEG) 4000. By selection using enzyme-linked immunosorbent assay (ELISA), three anti-CacyBP MAbs were obtained. The MAb BD1, whose isotype was IgG1, interacted with the fusion protein. Western blot and immunofluorescence microscopy showed that the MAb BD1 against CacyBP could recognize CacyBP protein derived from human gastric cancer cell lines in both native and denatured forms. This MAb would act as a useful tool for the detection of CacyBP protein in future studies.

Objective Calcyclin-binding protein or Siah-1-interacting protein (CacyBP/SIP) is a target protein of the S100A6, which is a member of the S100 family. It has been shown that CacyBP/SIP plays important roles in cell cycle progression, differentiation, and carcinogenesis. The purpose of this study was to explore the correlation of CacyBP/SIP expression and clinical significance in breast cancer. Methods Immunohistochemistry was performed to detect the CacyBP/SIP expression in 79 pairs of breast cancer tissues and adjacent nontumor tissues. Western blot analysis and semiquantitative RT-PCR were used to measure CacyBP/SIP protein and mRNA levels in three pairs of fresh breast cancer tissues and adjacent nontumor tissues. The clinical data were collected by telephone follow-up for 5 years. Results CacyBP/SIP mRNA and protein levels in breast cancer tissues were significantly higher compared with adjacent nontumor tissues. Poor cellular differentiation, lymph node invasion, and clinicopathological staging in breast cancer were associated with CacyBP/SIP expression. Other factors, such as age, menses, and tumor size, were not related to CacyBP/SIP expression. The up-regulation of CacyBP/SIP expression also was shown to increase recurrence and metastasis of breast cancer and to correlate with short overall survival rate. Conclusions CacyBP/SIP expression is evident in more advanced clinical progression of breast cancer, and it might be a biomarker for poor prognosis and a potential therapeutic target in breast cancer.

10 páginas, 6 figuras, 2 tablas., MC3 is a colorectal cancer (CRC)-specific mAb previously prepared in our laboratory that can detect CRC with high sensitivity and specificity. However, the target antigen for MC3 had not been identified due to technological limitations. In the present study, immunocytochemistry and immunohistochemistry revealed the expression patterns of MC3 antigen (MC3-Ag) in colon cancer cell lines and CRC tissues. Western blotting analysis showed that the MC3 antibody reproducibly recognized two ∼30 kDa proteins in the total cell lysates of human colon carcinoma cell lines SW480 and HT-29. Using a proteomic approach, we identified two MC3 immunoreactive spots as two isoforms of thioredoxin-like 2 (Txl-2) protein. Further paired immunostaining showed that Txl-2 had the same expression profile as probed by the MC3 antibody. Western blotting also showed that both antibodies could detect the same two bands, further verifying that Txl-2 is the antigen of MC3 antibody. Additionally, tissue arrays revealed the expression patterns of Txl-2 in various normal and cancer tissues. Further analysis showed that Txl-2 mRNA was elevated in 18 cases of CRC tissues compared to paracancerous tissues and adjacent normal tissues., This work was supported by grants from National Natural Science Foundation of China (No. 30672399) and National High Technology Research and Development Program of China (No. 2006AA02A253).

Calcyclin-binding protein (CacyBP)/Siah-1 interacting protein (SIP), a component of ubiquitin-mediated proteolysis, could bind the Skp1-Cul1-F box protein complex. Although CacyBP/SIP was implicated in p53-induced β-catenin degradation, its exact function was still unknown. Our previous studies showed that CacyBP/SIP could modulate the multidrug-resistant phenotype of gastric cancer cells and was highly expressed in gastric cancer tissues compared with that in non-cancerous tissues. In this study, CacyBP/SIP protein expression profile in a broad range of human normal tissues and carcinomas was analyzed by immunohistochemistry staining with anti-CacyBP/SIP monoclonal antibody first produced in our laboratory. CacyBP/SIP was generally localized in the cytoplasm/nucleus. Positive staining of CacyBP/SIP was found in brain, heart, lymph node, and esophagus. Weak staining was shown in the rectum and kidney. No CacyBP/SIP was detected in other normal tissues. However, CacyBP/SIP was ubiquitously detected in all kinds of tumor tissues and was highly expressed in nasopharyngeal carcinoma, osteogenic sarcoma, and pancreatic cancer. To our knowledge, this is the first study on the CacyBP/SIP expression pattern in a broad range of human normal and tumor tissues. The data presented should serve as a useful reference for other investigators in future studies of CacyBP/SIP functions. Hopefully, this knowledge will lead to discovery of more roles of CacyBP/SIP in tumorigenesis. (J Histochem Cytochem 56:765–772, 2008)

To explore the proteins regulated by cyclooxygenase-2 (COX-2) in gastric cancer, the expression plasmid of COX-2siRNA was constructed and transfected into gastric cancer cell line SGC7901. Then, two-dimensional electrophoresis and the PDQuest software analysis were applied to discover the differentially expressed proteins. The differential protein spots were analyzed by matrix-assisted laser desorption/ionization time of flight mass spectrometry. Fourteen differentially expressed proteins between the two cell lines were identified. 15-Hydroxyprostaglandin dehydrogenase [NAD(+)] (15-PGDH), a key enzyme in prostaglandin degradation, was identified as an upregulated protein in SGC7901 cells transfected with the COX-2siRNA plasmid. To further explore whether the 15-PGDH is regulated by COX-2, western blotting and immunocytochemical assay were performed to detect the expression of 15-PGDH in different cell lines with different expression level of COX-2. The results showed that the expression of 15-PGDH was upregulated (128.57%) as COX-2 was suppressed by small interfering RNA and downregulated (51.72%) as COX-2 was enhanced by COX-2 cDNA transfection in gastric cancer cells. In tissue specimens with gastric cancer, there was a decreased expression of 15-PGDH and an increased expression of COX-2 simultaneously. A significantly negative correlation of 15-PGDH expression was found to COX-2 level, tumor differentiation, tumor, lymph node, metastasis (TNM) staging and lymph node metastasis of gastric cancer. All the results suggest that 15-PGDH is downregulated by COX-2 in human gastric cancer and may contribute to the carcinogenesis and development of human gastric cancer in combination with COX-2.

Investigations of the odontoblast phenotype are hindered by obstacles such as the limited number of odontoblasts within the dental pulp and the difficulty in purification of these cells. Therefore, it is necessary to develop a cell culture system in which the local environment is inductive and can promote dental pulp stem cells (DPSCs) to differentiate into odontoblast lineage. In this study, we investigated the effect of conditioned medium from developing tooth germ cells (TGCs) on the differentiation and dentinogenesis of DPSCs both in vitro and in vivo. DPSCs were enzymatically isolated from the lower incisors of 4-week-old Sprague-Dawley rats and co-cultured with TGC conditioned medium (TGC-CM). The cell phenotype of induced DPSCs presents many features of odontoblasts, as assessed by the morphologic appearance, cell cycle modification, increased alkaline phosphatase level, synthesis of dentin sialoprotein, type I collagen and several other noncollagenous proteins, expression of the dentin sialophosphoprotein and dentin matrix protein 1 genes, and the formation of mineralized nodules in vitro. The induced DPSC pellets in vivo generated a regular-shaped dentin-pulp complex containing distinct dentinal tubules and predentin, while untreated pellets spontaneously differentiated into bone-like tissues. To our knowledge, this is the first study to mimic the dentinogenic microenvironment from TGCs in vitro, and our data suggest that TGC-CM creates the most odontogenic microenvironment, a feature essential and effective for the regular dentinogenesis mediated by DPSCs.

Aim of the Study Croton tiglium (Croton tiglium L., Euphorbiaceae) is widely used as a herb for treatment of gastrointestinal disturbances. Previous studies established its purgative and inflammational properties. The present study aimed to investigate the effects of Croton tiglium oil (CO) on intestinal transit in mice. Materials and Methods Gastrointestinal transit in mice and contractile characteristics of isolated intestinal strips from mice were evaluated. Intestinal inflammation was confirmed by histological examination. Results Low dose of CO increased the gastrointestinal transit of charcoal and barium meal as well as the production of fecal pellets in mice. In contrast, high dose exerted inhibitory effects. For normal colonic circular strips, both high and low dose of CO inhibited the contractile frequency. Low doses (0–20 μg/ml) of CO enhanced the phasic contractions, while high doses (>40 μg/ml) reduced them. Colonic longitudinal strips in CO-treated mice were less sensitive to electrical field stimulation than those in control mice. The contraction of colonic longitudinal, colonic and jejunal circular strips in CO-treated mice was more sensitive to atropine than that in control mice. Conclusions CO might modulate gastrointestinal motility and induce intestinal inflammation related to immunological milieu and motor activity. Our findings may highlight the ethno-medical uses of Croton tiglium on intestinal disorders.

Estrogen responsiveness of bone formation is mediated by the estrogen receptor alpha (ERalpha) in osteoblastic lineage. As osteoblasts arise from the multipotent bone marrow stromal (mesenchymal) cells, this study was undertaken to observe the ERalpha in primary female adult rat bone marrow mesenchymal stem cells (BMSCs). The ERalpha was localized using immunocytochemical analysis in identified primary BMSCs. Then, using real-time PCR analysis, we measured the expression of ERalpha messenger RNA (mRNA) in BMSCs. ERalpha transcripts showed different trends between untreated cultures (control group) and osteogenic-induced cultures (treated group). In the control group, ERalpha mRNA climbed at peak levels at a confluence stage and decreased until day 20, whereas, in the treated group, the ERalpha mRNA kept climbing from a low level until day 20. Thus, the observed developmental expression of ERalpha mRNA correlates with progressive BMSCs growth and osteogenic differentiation and BMSCs may be a primary target cell for estrogen in maintaining bone formation.

The aim of this study was to establish an orthotopic implantation model with high metastasis of gastric cancer to the peritoneum which is more faithful to clinical metastasis. A human gastric carcinoma cell line, GC9811, was injected as a single-cell suspension into the stomach of nude mice. The cells from some peritoneum metastatic foci were expanded in vitro and subsequently implanted to the stomach wall of nude mice. By repeating the in vivo stepwise selection method for four rounds and cloning culture, we obtained a cell line designated GC9811-P, which developed peritoneal metastasis in 13 of 13 (100%) of mice, compared with only 20% of those implanted with parental GC9811. The metastatic foci in the peritoneum showed essentially the same histological appearance as those induced by parental cells. Tumor cell growth of GC9811-P in vitro was faster than that of GC9811. Motility assays demonstrated higher motility of GC9811-P than of GC9811. The adhesive ability of GC9811-P cells to laminin was lower than that of GC9811 cells, whereas the ability of GC9811-P cells to adhere to fibronectin was significantly higher than that of parental cells. Differences between GC9811-P and their parental GC9811 cells were found in expression levels of various molecules by flow cytometric and western blot. The findings indicated that up-regulation in the expressions of CD155, VEGF, syndecan-1, and syndecan-2 or down-regulation in the expressions of IL-6 and E-cadherin play an important role in the peritoneal metastasis of human gastric carcinoma cells. The high-metastatic cell line appears to be useful for investigating the mechanisms of peritoneal metastasis and preventing peritoneal metastasis of human gastric cancer.

We investigated the potential effect of Cyclooxygenase-2 (Cox-2) on hypoxia-induced Angiopoietin-2 (Ang-2) expression in gastric cancer cells. Our results revealed that hypoxia augmented Cox-2 and Ang-2 expressions. Also, the hypoxia-induced Ang-2 could be mimicked by CoCl(2) treatment while genestein treatment could partially counteract the hypoxia-induced Ang-2 expression. Celecoxib but not Cox-1 inhibitor sc-560 reversed the hypoxia-induced Ang-2 expression, while this effect could be partially restored by addition of exogenous PGE2. Our findings suggest that the hypoxia-elevated Ang-2 expression in gastric cancer cells may be mediated by both Cox-2-derived PGE2 and HIF-1alpha pathways, while celecoxib could counteract the hypoxia-induced Ang-2 expression.

Ribosomal proteins (RP) S13 and RPL23 were previously identified as two upregulated genes in a multidrug-resistant gastric cancer cell line SGC7901/VCR compared to its parental cell SGC7901 by differential display PCR. The aim of this study was to explore the roles of RPS13 and RPL23 in multidrug resistance (MDR) in gastric cancer cells. RPS13 and RPL23 were genetically overexpressed in SGC7901 cells, respectively. Either RPS13 or RPL23 enhanced resistance of SGC7901 cells to vincristine, adriamycin, and 5-fludrouracil. RPL23 also enhanced resistance of SGC7901 cells to cisplatin. Overexpression of either RPS13 or RPL23 did not alter the population doubling time, [3H]leucine incorporation, and intracellular adriamycin accumulation of SGC7901 cells. However, either RPS13 or RPL23 could protect SGC7901 cells from undergoing vincristine-induced apoptosis. Western blot analysis revealed that both RPS13 and RPL23 significantly increased the expression level of Bcl-2 and Bcl-2/Bax ratio in SGC7901 cells. In addition, overexpression of RPL23 enhanced glutathione S-transferase (GST) activity and intracellular glutathione content in SGC7901 cells. Together, this work demonstrates that either RPS13 or RPL23 can promote MDR in gastric cancer cells by suppressing drug-induced apoptosis, and that RPL23 may also promote MDR in gastric cancer cells through regulation of glutathione S-transferase-mediated drug-detoxifying system.

We report the isolation and functional characterization of the gene encoding MGr1-Ag, a multidrug-resistance-associated protein. A lambdagt11 cDNA library derived from colorectal carcinoma SW480 cells was screened with monoclonal antibody MGr1. DNA homology analysis of 22 positive clones (designated R1-R22) suggested human 37-kDa laminin receptor precursor (37LRP, R7/R9/R15/R16/R19/R20) and a novel gene (R22) as candidate genes encoding MGr1-Ag. Western blot analysis showed that anti-R20 serum reacted with a unique protein band that was consistent with MGr1-Ag, while anti-R22 serum could not react with MGr1-Ag. The coding gene for MGr1-Ag was amplified using reverse transcription-PCR. Sequence analysis revealed that the MGr1-Ag and 37LRP genes shared the same coding sequence. An in vitro drug sensitivity assay indicated that down-regulation of 37LRP by an antisense technique could significantly enhance the cytotoxicity of anticancer drugs to gastric cancer cells. Thus we draw the conclusion that MGr1-Ag is identical to 37LRP.

To study the mechanisms of multidrug resistance (MDR) mediated by human 67 000 laminin receptor (LR) with a relative molecular mas of 67 000 in gastric cancer cells.Antisense RNA expression vector corresponding to LR precursor (LRP) was constructed by DNA recombinant technique, and transferred into gastric cancer MDR cells SGC7901/VCR with Lipofect AMINE. Western blot was employed to determine the LR expression level in gastric cancer cells. The sensitivity of gastric cancer cells to chemotherapeutic drugs was evaluated with MTT assay. Flow cytometry was used to analyze the cell cycle and to assess the mean fluorescence intensity of intracellular adriamycin in gastric cancer cells.Western blotting analysis demonstrated a decreased expression level of LR in SGC7901/VCR cells transfected with LRP antisense RNA expression vector. In comparison with the gastric cancer cells with out transfection or transfected with invalid vector, LR down-regulated transfectants (SGC7901/VCR-anLRP) showed higher sensitivity to vincristine, adriamycin, 5-fluodrouracil and cisplatin, and increased accumulation and retention of adriamycin. Cell cycle analysis suggested G1 block and spontaneous apoptosis of SGC7901/VCR-anLRP cells.LR might take part in mediation of MDR in gastric cancer cells through interfering with drug accumulation and cell apoptosis.

Background: Dendritic cells (DC) and macrophages (Mφ) mediate intestinal immune tolerance. Recent evidence suggests intestinal CD103+CD11b- DC are involved in generation of regulatory T cells (T-reg), with CD103+CD11b+ DC stimulating Th17 cell differentiation. Information regarding gut DC function during inflammation is scarce. Methods: We compared murine and human gut DC and Mφ in the steady state and during intestinal inflammation, by flow cytometry. Results: Proportions of CD103-CD64+ Mφ were enhanced in the inflamed IL-10 knockout (KO) murine colon compared to WT, alongside a loss of CD103+CD11b- DC. Proportions of CD4+FoxP3+ T-reg were reduced in the inflamed IL-10 KO colon, with enhanced production of IL-17A and TNFα by T cells but a loss of regulatory TGFβ production by CD103+ DC. Proportions of CD103-CD64+ Mφ were also enhanced in the human Crohn’s disease colon compared to non-inflamed control tissue (diverticulitis), alongside a loss of CD103+ DC. CD103 was co-expressed with either CD141 or CD1c on human intestinal DC, corresponding to CD103+CD11b- and CD103+CD11b+ murine DC respectively. Proportions of all CD103+ gut DC subsets were reduced in human Crohn’s disease. Conclusions: Lack of IL-10 may skew DC and Mφ differentiation in the gut, leading to loss of DC able to generate T-reg. Further experiments will determine the ability of intestinal DC and Mφ subsets to stimulate specific T cell responses in vitro and the roles of IL-10 and TGFβ during this process.