黄芩苷通过调控c-Myc介导的细胞周期抑制胆管癌细胞增殖  

Baicalin inhibits human cholangiocarcinoma cell proliferation via regulating c-Myc mediated cell cycle

作　　者：徐杨 肖斌 魏嵋 代荣阳 向远彩 XU Yang;XIAO Bin;WEI Mei;DAI Rong-yang;XIANG Yuan-cai(Department of Biochemistry and Molecular Biology,Southwest Medical University,Luzhou 646000,China;Department of Liver Diseases,the Affiliated Hospital of Chinese Traditional Medicine Southwest Medical University)

机构地区：[1]西南医科大学生物化学与分子生物学教研室,646000 [2]西南医科大学附属中医医院肝病科

出　　处：《天津医药》2020年第8期705-710,共6页Tianjin Medical Journal

基　　金：四川省教育厅创新团队(16TD0021);泸州市西南医科大学联合基金(2019LZXNYDZ03);四川科技厅基金(2019YJ0482)。

摘　　要：目的探究黄芩苷(BC)对人胆管癌(CCA)细胞QBC939和RBE增殖的影响及其潜在机制。方法取对数生长期QBC939和RBE细胞,不同浓度的BC处理细胞24 h后,采用CCK-8法分别检测BC对CCA细胞增殖及敲低原癌基因蛋白质(c-Myc)后对BC引起细胞增殖活性改变的影响;高倍显微镜观察BC对细胞生长状态的影响;流式细胞术检测BC对CCA细胞周期的影响;蛋白免疫印迹(Western blot)实验分别检测BC及小干扰RNA(siRNA)敲低CCA细胞中c-Myc后对周期相关蛋白周期素依赖激酶抑制剂(p27)、细胞周期蛋白D1(Cyclin D1)及c-Myc表达的影响。结果与0μmol/L组相比,BC明显抑制QBC939和RBE细胞的增殖活性(P<0.01),且高倍显微镜下观察到细胞生长减缓;BC可将QBC939细胞周期阻滞于S期(P<0.05);随着BC浓度的增加,p27的蛋白水平明显上调,而Cyclin D1则显著降低(P<0.01);BC可下调c-Myc的蛋白表达,且敲低c-Myc后p27和Cyclin D1蛋白表达的变化趋势与BC处理时一致(P<0.05);单独敲低c-Myc后可明显抑制QBC939和RBE细胞的存活率(P<0.01),并能进一步增强BC的抑制作用。结论BC通过抑制c-Myc信号通路阻滞细胞周期,进而抑制CCA细胞的增殖。Objective To investigate the effect of baicalin(BC)on the proliferation of human cholangiocarcinoma(CCA)cell lines,QBC939 and RBE,and its underlying mechanism thereof.Methods QBC939 and RBE cells on logarithmic growth phase were treated with different concentrations of BC for 24 h,and CCK-8 was used to evaluate the effect of BC alone treatment or in combination with c-Myc knockdown on the cell proliferation activity.The effect of BC on cell growth status was observed by microscope in CCA cells.The effect of BC on the cell cycle was examined by flow cytometry.Western blot assay was also employed to detect the effects of BC and c-Myc knockdown mediated by siRNA on the expressions of cell cycle related proteins,such as cyclin-dependent kinase inhibitor p27,Cyclin D1 and proto-oncogene protein c-Myc,respectively.Results BC significantly inhibited the proliferation activities of QBC939 and RBE cells compared with 0μmol/L group(P<0.01).BC-treated cells were slow growth observed under the microscopic.BC significantly induced cell cycle arrest in S phase in QBC939 cells(P<0.05).The protein levels of p27 and Cyclin D1 were markedly increased and decreased,respectively with the increase of BC concentrations(P<0.01).The protein level of c-Myc was also declined after BC treatment.The knockdown of c-Myc alone with siRNAs demonstrated a same change in the protein levels of p27 and Cyclin D1 compared to BC treatment in QBC939 and RBE cells(P<0.05).The knockdown of c-Myc alone can significantly inhibit the proliferation activities of QBC939 and RBE cells and further enhance the inhibitory effect of BC(P<0.01).Conclusion BC inhibits the proliferation of CCA cells via blocking c-Myc signaling pathway.

关 键 词：黄芩甙 细胞增殖 细胞周期 周期素依赖激酶抑制剂P27 细胞周期蛋白D1 原癌基因蛋白质C-MYC 胆管肿瘤 RNA 小分子干扰 

分 类 号：R735.8[医药卫生—肿瘤] R349.53[医药卫生—临床医学]