摘要
目的观察瘦素对人胆管癌细胞QBC939增殖和凋亡的影响并初步探讨其可能的机制。方法在培养液中加入不同浓度的瘦素后,用四甲基偶氮唑盐(MTT)法检测细胞增殖,流式细胞仪观察细胞周期及凋亡情况,实时荧光定量PCR法检测增殖相关基因Cyclin D1和凋亡相关基因Bax、Bcl-2的表达情况,同时检测Caspase-3的活性。结果 MTT法显示瘦素可以促进QBC939细胞的增殖;流式细胞仪检测结果显示瘦素能明显降低G0/G1期细胞比例并提高S期细胞比例,细胞凋亡率也有明显降低;实时荧光定量PCR法显示瘦素(50 ng/ml)处理QBC939细胞24 h后,Cyclin D1mRNA的表达明显增高,Bcl-2 mRNA的表达量明显增高,而Bax mRNA的表达量下降,差异有统计学意义;而且瘦素作用QBC939细胞后能降低细胞的Caspase-3酶活性。结论瘦素可以明显的促进人胆管癌细胞QBC939从细胞周期G0/G1期向S期转换,进而促进细胞增殖,此外瘦素还可以抑制其凋亡。
Objective To determine whether leptin can exert anti-proliferative and pro-apoptotic effects on human cholangiocarcinoma cells and to investigate the underlying molecular mechanisms.Methods Human cholangiocarcinoma QBC939 cells were cultured and treated with different concentrations of leptin.Changes in the proliferation rate were measured by the MTT assay.Changes in cell cycle and in the apoptosis incidence rate were detected by flow cytometry.Changes in cyclin D1,bax and bcl-2 gene expression were detected by measuring mRNA levels by real-time quantitative reverse transcription-polymerase chain reaction（qPCR）.Changes in caspase-3 protease activity were detected by fluorometric assay.Results Leptin treatment significantly increased the proliferation rate of QBC939 cells in a dose-and time-dependent manner.Compared to untreated QBC939 cells,leptin treatment led to significantly more G0/G1 to S phase transition and significantly lower apoptosis rate.In addition,leptin-treated QBC939 cells showed enhanced mRNA expression of cyclin D1 and bcl-2,but decreased mRNA expression of bax.The leptin treatment also led to decreased caspase-3 activity.Conclusion Leptin promotes S to G0/G1 phase transition and proliferation,but inhibits apoptosis,of human cholangiocarcinoma cells in vitro.
出处
《临床肝胆病杂志》
CAS
2013年第3期223-226,共4页
Journal of Clinical Hepatology
基金
湖北省自然科学基金(2008CDB200)
武汉大学自主科研项目(自然科学)(111125)
关键词
胆管肿瘤
瘦素
增殖
凋亡
bile duct neoplasms
leptin
proliferation
apoptosis
