c-AMP/PKA信号通路在γ-氨基丁酸抑制胆管癌细胞株QBC939生长中的作用研究  

The effect of cAMP / PKA signaling pathway in γ-aminobutyric acid inhibitory on the growth of cholangiocarcinoma QBC939 cell line

作　　者：王成 黄强 朱成林 黄朝君 刘臣海 谢放 王伟[2] 

机构地区：[1]安徽医科大学附属省立医院普外科胆胰病区肝胆胰外科安徽省重点实验室,合肥230001 [2]安徽医科大学附属省立医院肿瘤化疗科

出　　处：《肝胆外科杂志》2015年第4期304-307,共4页Journal of Hepatobiliary Surgery

基　　金：安徽省科技攻关项目(编号:1301043042)

摘　　要：目的探讨c-AMP/PKA信号通路在γ-氨基丁酸(GABA)抑制胆管癌细胞株QBC939生长中发挥的作用。方法分别以GABA、GABA+8Br(c AMP激动剂)、GABA+H89(PKA拮抗剂)孵育胆管癌细胞株QBC939 48 h,采用四甲基偶氮唑蓝(MTT)法及Annexin V-FITC/PI流式细胞术检测细胞的增殖及凋亡,ELISA法检测c AMP、蛋白激酶A(PKA)表达情况,Western blot法检测PKA I/PKA II蛋白表达。结果 MTT及流式细胞术检测结果显示8Br协同GABA所导致的增值抑制(20.30%±0.57%vs 16.11%±0.73%,t=8.977,P<0.05)和凋亡效应(32.62%±2.45%vs 28.38%±1.53%,t=2.929,P<0.05),而H89拮抗此效应;ELISA检测结果显示GABA明显上调c AMP及PKA的含量,8Br协同GABA增加c AMP含量(26.08±0.15 vs 25.88±0.06,t=8.565,P<0.05),H89拮抗GABA所导致PKA的含量(933.13±3.13 vs 947.71±6.51 t=2.512,P<0.05);Western blot检测结果显示,GABA较对照组能明显下调PKA I(0.0878±0.003 vs.0.1521±0.003,t=29.687,P<0.05),上调PKA II蛋白的表达(0.2042±0.012 vs 0.1461±0.072,t=8.152,P<0.05)。结论 GABA可能通过c AMP/PKAII信号通路发挥抑制胆管癌细胞株QBC939生长的作用。Objective To explore the effect of c AMP/PKA signaling pathway in γ-aminobutyric acid inhibitory on the growth of cholangiocarcinoma QBC939 cell line. Methods QBC939 cells were cultured in different groups and treated with GABA,GABA +8Br( c AMP agonists) 、GABA + H89( PKA antagonist) for 48 hours. MTT assay was used to determine the proliferation of QBC939 cells. Annexin V-FITC / PI binding assay was used to detect apoptosis in the QBC939 cells. ELISA assay was detected to the expression of c AMP and PKA. Western blot was applied to check the expression of PKAI and PKAII proteins in different groups of QBC939 cells.Resutls MTT and FCM assays all showed that the effect of GABA inhibitory on the proliferation( 20. 30% ± 0. 57% vs. 16. 11% ±0. 73%,t = 8. 977,P < 0. 05) and induced apoptosis( 32. 62% ± 2. 45% vs. 28. 38% ± 1. 53%,t = 2. 929,P < 0. 05) of QBC939 cells could be enhanced by 8Br,but not H89. GABA significantly increased the content of c AMP and PKA,the content of c AMP was enhanced by 8Br( 26. 08 ± 0. 15 vs. 25. 88 ± 0. 06,t = 8. 565,P < 0. 05),the content of PKA was antagonized by H89( 933. 13 ±3. 13 vs. 947. 71 ± 6. 51 t = 2. 512,P < 0. 05). Western blot analysis showed that GABA significantly down-regulated the expression of PAK I protein( 0. 0878 ± 0. 003 vs. 0. 1521 ± 0. 003,t = 29. 687,P < 0. 05),and up-regulated the expression of PKA II protein( 0. 2042 ± 0. 012 vs. 0. 1461 ± 0. 072,t = 8. 152,P < 0. 05). Conclusions GABA may inhibit the growth of cholangiocarcinoma cells QBC939 through c AMP / PKAII signaling pathway.

关 键 词：胆管肿瘤 Γ-氨基丁酸 环磷酸腺苷 蛋白激酶A 

分 类 号：R735.8[医药卫生—肿瘤]