多西环素对体外人小细胞肺癌H446细胞增殖的影响及其机制  

Effect of doxycycline on the proliferation of human small cell lung cancer H446 cells in vitro and its mechanisms

作　　者：张健明 司徒伟勤 宋志华[2] 王胜奇[2] 

机构地区：[1]开平市第二人民医院,开平529367 [2]南方医科大学南方医院,广州510515

出　　处：《国际药学研究杂志》2017年第1期47-51,共5页Journal of International Pharmaceutical Research

基　　金：国家自然科学基金资助项目(81173014);广东省科技计划资助项目(2014A020212197)

摘　　要：目的探讨多西环素对体外人小细胞肺癌H446细胞增殖的影响及其可能的作用机制。方法不同浓度的多西环素作用于体外培养的人小细胞肺癌H446细胞24、48、72或96 h,采用CCK-8法检测多西环素对H446细胞增殖的影响;采用Bliss法计算半数抑制浓度(IC50);采用TUNEL法检测多西环素对H446细胞凋亡的影响;采用Western印迹法检测多西环素对Bcl-2、Bax等凋亡相关蛋白表达的影响;采用实时定量-PCR法检测多西环素对Bcl-2、Bax等凋亡相关基因m RNA表达的影响。结果多西环素可浓度-时间依赖性地抑制人小细胞肺癌H446细胞的体外增殖,作用24、48、72和96 h对应的IC50值分别为24.10、10.98、8.20和8.03 mg/L;多西环素作用后H446细胞的凋亡指数显著增加(P<0.05);Bax的蛋白及m RNA表达水平上调,Bcl-2的蛋白及m RNA表达水平下调,差异具有统计学意义(P<0.05)。结论多西环素可通过上调促凋亡相关蛋白Bax的表达,下调抑凋亡相关蛋白Bcl-2的表达,诱导人小细胞肺癌H446细胞凋亡,进而抑制人小细胞肺癌H446细胞的体外增殖,是一种有开发前景的小细胞肺癌治疗药物。Objective To investigate the effect of doxycycline on the proliferation of human small cell lung cancer H446 cells in vitro and its possible mechanisms. Methods H446 cells were treated with different concentrations of doxycycline for 24,48,72 or96 h. Cell proliferation was measured by CCK-8 assay,IC50 was caculated by Bliss,and cell apoptosis was examined by TUNEL assay.The protein and m RNA expression of Bcl-2 and Bax in H446 cells were detected by Western blot and real time-PCR,respectively. Results Doxycycline inhibited the proliferation of H446 cells in a concentration-time dependent manner. The half inhibitory concentrations（IC50）of doxycycline on cell proliferation in H446 cells were approximately 24.10 mg/L for 24 h treatment group,10.98 mg/L for48 h treatment group,8.20 mg/L for 72 h treatment group and 8.03 mg/L for 96 h treatment group,respectively. Furthermore,doxycycline treatment could also induce the apoptosis of H446 cells in a concentration-dependent manner. Moreover,doxycycline treatment dramatically down-regulated the expression of Bcl-2 and up-regulated the expression of Bax in H446 cells both at the protein and at the m RNA levels. The differences were statistically significant（P〈0.05）. Conclusion Doxycycline can inhibit the proliferation of H446 cells by inducing apoptosis via down-regulating the expression of Bcl-2 and up-regulating the expression of Bax. Doxycycline has great potential as a chemotherapeutic agent for human small cell lung cancer.

关 键 词：多西环素 小细胞肺癌 增殖 凋亡 药物疗法 

分 类 号：R734.2[医药卫生—肿瘤]