雷帕霉素、吉西他滨、阿霉素联合化疗对人胆管癌HCCC-9810细胞中Survivin、VEGF-C、p-mTOR、mTOR蛋白表达的影响

Effect of rapamycin and doxorubicin, gemcitabine combined with chemotherapy on human cholangiocarcinoma HCCC-9810 cells in Survivin, VEGF-C, p-mTOR, mTOR protein

目的 Rapamycin,探讨雷帕霉素(RAP)、吉西他滨(Gemcitabine,GEM)、阿霉素(Doxorubicin,DOX)联合化疗对人胆管癌HCCC-9810细胞中Survivin、VEGF-C、p-m TOR、m TOR蛋白表达的影响。随机分为对照组、方法 RAP组、GEM组、DOX组、RAP+GEM组、RAP+DOX组、GEM+DOX组、RAP+GEM+DOX组。MTT实验界定3种药物的干预浓度。Western Blot技术检测3种药物联合用药对胆管癌细胞系HCCC-9810细胞中Survivin、VEGF-C、p-m TOR、m TOR蛋白表达的影响。结果 RAP、GEM和DOX的干预浓度分别为25 nmol/L、50μmol/L和1μmol/L。RAP(25 nmol/L)、GEM(50μmol/L)和DOX(1μmol/L)对HCCC-9810细胞增殖具有4.19%协同抑制作用。RAP、RAP+GEM、GEM+DOX RAP+、GEM+DOX能显著下调胆管癌HCCC-9810细胞中Survivin蛋白水平(P<0.01);RAP、DOX、GEM+DOX、RAP+DOX、RAP+GEM+DOX能显著下调VEGF-C蛋白水平(P<0.01或0.05);RAP、GEM、DOX、RAP+GEM、RAP+DOX RAP+、GEM+DOX能显著下调p-m TOR蛋白水平(P<0.01或0.05);3种药物均对m TOR蛋白表达无影响(P>0.05)结论。RAP、GEM、DOX联合用药能明显抑制Survivin、VEGF-C和p-m TOR蛋白的表达,而对m TOR蛋白的表达无明显抑制。

Objective To explore the influence on expression of four proteins Survivin, VEGF-C, p-m TOR, m TOR by the combination treatment of rapamycin（RAP）, gemcitabine（GEM） and doxorubicin（DOX） in human cholangiocarcinoma HCCC-9810 cells. Methods To define three kinds of drug intervention concentration determined by MTT experiment; Group of Experiments： control group, RAP, GEM, DOX, RAP＋GEM, RAP＋DOX, GEM＋DOX, RAP＋GEM＋DOX groups. To detect the influence of Survivin, VEGF-C, p-m TOR, m TOR protein by the combination treatment of RAP, GEM and DOX in the HCCC-9810 cells of human cholangiocarcinoma with Western Blotting. Results The concentrations of combination treatment groups of RAP, GEM, DOX were 25 nmol/L, 50 μmol/L, 1 μmol/L respectively. RAP＋DOX significantly downregulated p-m TOR protein expression（P〈0.01）, group GEM＋DOX significantly reduced Survivin, VEGF-C protein expression（P〈0.01）, group RAP＋GEM＋DOX significantly reduced Survivin, p-m TOR, VEGF-C protein expression（P〈0.01 or 0.05）. Conclusion RAP, GEM, DOX combination can significantly inhibit the expression of VEGF-C, p-m TOR and Survivin protein, but the expression of protein m TOR in no significant inhibition.