To Investigate The Mechanism And Correlation Effect Genes Of γ-aminobutyric Acid In The Cholangiocarcinoma Cells

Object To investigate the mechanism and correlation effect genes of γ‐ aminobutyric acid in the cholangiocarcinoma cells.Methods The first,43 cases tissue samples of cholangiocarcinoma were collected, and 12 cases of normal bile duct tissues as controls. The tissues were examined for the expression of stat3, p‐stat3, survivin and cox‐2 by immunohistochemistry. The second,QBC939 cholangiocarcinoma cell line cultures were divided into experimental group, GABA treatment group, GABA + phaclofen(B receptor antagonist) group, AG490(JAK / STAT3 signaling pathway antagonist) group. MTT assay was used to check the proliferation activity, flow cytometry to detect apoptosis. Immunohistochemistry, Western blotting, RT‐PCR testingwere used to detect the expression of p‐stat3, Survivin and COX‐2 protein. The third, The QBC939 cell was inoculated in nude mice to cultures xenograft model. They were divided into GABA treatment group and the control group, the immunohistochemistry and WB were used to test each protein assay.Results With the results of the positive rate of stat3, p‐stat3, survivin and cox‐2 incholangiocarcinoma was 69.8%(30/43)、65.1%(28/43)、72.1%(31/43)、79.1%(34/43), meanwhile it was 1.7%(5/12)、8.3%(1/12)、16.7%(2/12)、41.7%(5/12)in normal bile duct. the differences in the expression of p‐stat3, survivin and cox‐2 protein in two groups were statistically significant(P <0.05). The QBC939 cell lines of GABA group and AG490 group contrast with the control group, tumor cell proliferation was decreased, and apoptosis rate was increased. With the application of phaclofen, tumor cell proliferation and apoptosis rate contrast with the control group was not statistically significant(p <0.05). The expression of p‐STAT3, Survivin and COX‐2 protein in GABA and AG490 control group by WB, RT‐PCR and immunohistochemistry compared with the control group was significantly decreased, the difference was statistically significant(p <0.05). Compared with the control group, each protein of transplanted tumors detected by the WB and immunohistochemistry in treatment group was significantly decreased, and the difference was statistically significant.Conclusion It may be one of the mechanisms that GABA inhibit the growth of cholangiocarcinoma QBCQ939 cell lines by mediated JAK / STAT3 signaling pathway, and reduced the expression of Survivin and COX‐2.