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Objective To investigate the combined effects of somatostatin (SST) and cisplatin (DDP) on proliferation and apoptosis in gallbladder cancer cells,and to investigate the mechanism of the combined effects.Methods We performed immunohistochemistry to detect the PTEN expression in gallbladder cancer.We then investigated the combined effects of SST and DDP on cell proliferation in vitro with CCK-8 assay and analyzed the interaction between these two drugs using isobologram analysis.We also investigated the combined effects on cell proliferation in vivo using a xenograft nude mouse model.FITC-Annexin V/PI assay and TUNEL staining assay were performed to detect the proportion of apoptosis after combined treatment in vitro and in vivo.Reactive oxygen species and mitochondrial membrane potential were detected with DCFH-DA assay and JC-1 staining assay after the combined treatment.We finally detected the PTEN and p-AKT associated proteins using western blotting after the combined treatment.Results PTEN was abnormally decreased in gallbladder cancer tissues.PTEN expression was negatively correlated with cancer differentiation and was positively correlated with patients'survival time.DDP treatment decreased while combined treatment with SST induced PTEN expression and inhibited AKT activation by reversing resistance to DDP.Isolated SST or DDP treatment inhibited gallbladder cancer GBC-SD and SGC996 cell proliferation which was dose-dependence.These two drugs synergistically inhibited gallbladder cancer cell growth in vivo and in vitro.Isolated SST or DDP treatment induced cell apoptosis and combined treatment induced cell apoptosis the most.SST inhibited AKT activation but did not induce ROS.DDP induced ROS resulting in increased cell apoptosis.Either SST or DDP alone increased the levels of cytoplasmic cytochrome C protein and activated caspase-3.Conclusions SST enhanced growth inhibition by cisplatin in gallbladder cancer cells through inducing PTEN expression.This study provides the theoretical basis for further combined clinical chemotherapeutic applications.
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Objective To investigate the predictive value of preoperative serum tumor markers test for lymph node metastasis of intrahepatic cholangiocarcinoma (ICC).Methods The retrospecgtive cohort study was conducted.The clinicopathological data of 69 patients with ICC who were admitted to the Xinhua Hospital of Shanghai Jiaotong University between May 2006 and May 2016 were collected.Among 69 patients with pathological diagnosis,24 with lymph node metastasis were allocated into the lymph node metastasis group and 45 without lymph node metastasis were allocated into the non-lymph node metastasis group.Tumor markers of the 2 groups were preoperatively detected,including alpha-fetoprotein (AFP),carcinoembryonic antigen (CEA),prostate specific antigen (PSA),CA19-9,CA125,CA242,CA153,CA724,CA211,neuron-specific enolase (NSE) and squamous cell carcinoma antigen (SCC).Receiver operating characteristic (ROC) curve was built,and critical value,sensitivity and specificity were calculated based on ROC curve.Coincident rate between significant indicators and results of pathological examination was calculated.Observation indicators:(1) overall positive rates of tumor markers;(2) comparison of tmmor markers levels in the 2 groups;(3) tumor markers predicted ROC curve of lymph node metastasis and coincident rate between ROC curve and results of postoperative pathological examination.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (Q25,Q75) and comparison between groups was analyzed using the Wilcoxon rank sum test.Comparison of count data was analyzed by the chi-square test and Fisher exact probability.The statistically significant indicators were analyzed by the ROC curve.Results (1) Overall positive rates of tumor markers:positive rates of AFP,CEA,PSA,CA19-9,CA125,CA242,CA153,CA724,CA211,NSE and SCC in 69 patients were 27.5％ (19/69),29.0％ (20/69),4.3％ (3/69),69.6％ (48/69),36.2％ (25/69),50.7％ (35/69),26.1％ (18/69),21.7％ (15/69),62.3％ (43/69),31.9％(22/69) and 21.7％(15/69),respectively.Positive rates of AFP,CEA,CA19-9,CA125,CA242,CA153,CA724,CA211,NSE and SCC were more than 20％,which became comparison indicators of 2 groups.(2) Comparison of tumor markers levels in the 2 groups:levels of CA19-9,CA125,CA242 and CA211 were 284.9 U/mL (42.5 U/mL,730.3 U/mL),63.6 U/mL (23.4 U/mL,172.1 U/mL),71.7 U/mL (25.6 U/mL,138.9 U/mL),6.7 μg/L (3.9 μg/L,17.5 μg/L) in the lymph node metastasis group and 58.0 U/mL (25.9 U/mL,405.9 U/mL),18.2 U/mL (11.7 U/mL,33.8 U/mL),11.0 U/mL (3.7 U/mL,41.7 U/mL),3.7 μg/L (2.7 μg/L,6.9 μg/L) in the non-lymph node metastasis group,respectively,with statistically significant differences between the 2 groups (Z=2.016,3.213,3.143,2.482,P＜0.05).(3) Tumor markers predicted ROC curve of lymph node metastasis and coincident rate between ROC curve and results of postoperative pathological examination:area under the ROC curve of CA19-9,CA125,CA242 and CA211 were respectively 0.648 [95％ confidence interval (C I):0.515-0.781,P＜0.05],0.736 (95％ CI:0.608-0.864,P＜0.05),0.731 (95％ CI:0.603-0.859,P＜0.05),0.714 (95％ CI:0.581-0.847,P＜0.05).The positive critical value,sensitivity and specificity of CA19-9,CA125,CA242 and CA21 were 150.6 U/mL,35.7 U/mL,43.4 U/mL,6.0 μg/L and 62.5％,66.7％,70.8％,62.5％ and 71.1％,82.2％,77.8％,75.6％,respectively.The coincident rate between ROC curve and results of postoperative pathological examination of CA 19-9,CA 125,CA242 and CA211 were 68.1％ (47/69),76.8％(53/69),75.4％(52/69),71.0％(49/69),respectively.Conclusion The levels of serum CA19-9,CA125,CA242 and CA211 can effectively predict lymph node metastasis of patients with ICC.