氧化苦参碱对HPV16感染宫颈癌Siha细胞增殖与凋亡的影响

Effect of Oxymatrine on the Proliferation and Apoptosis of Cervical Cancer Siha Cells Infected by HPV16

目的:探讨氧化苦参碱对人类乳头瘤病毒(human papilloma virus,HPV)16感染宫颈癌Siha细胞增殖与凋亡的影响。方法:以人子宫颈鳞癌Siha细胞作为HPV16感染型宫颈癌体外实验模型,常规复苏、培养及传代;加入不同质量浓度氧化苦参碱(0.2 g·L^-1、0.4 g·L^-1、0.8 g·L^-1、1.6 g·L^-1)作为实验组,加入等体积RPMI-1640作为溶剂对照组,作用24 h、48 h、72 h;噻唑蓝法检测细胞增殖活性改变,流式细胞仪分析细胞凋亡率,蛋白免疫印迹法分析凋亡相关蛋白Bcl-2、Bax、P53表达。结果:噻唑蓝结果显示:氧化苦参碱对HPV16感染宫颈癌Siha细胞增殖抑制作用呈质量浓度和时间依赖性(P<0.05)。流式细胞仪检测结果显示:不同质量浓度氧化苦参碱作用48 h后,细胞凋亡率与对照组比较,差异有统计学意义(P<0.05);氧化苦参碱作用48 h后,与对照组比较,实验组Siha细胞G0/G1期比率上升(P<0.05),S期和G2/M期比率下降(P<0.05),细胞生长周期停滞于G1期。蛋白免疫印迹检测结果显示:不同质量浓度氧化苦参碱作用48 h后,实验组凋亡相关蛋白Bcl-2蛋白表达下调,Bax、P53蛋白表达上调,且其上调(下调)呈质量浓度依赖性。结论:氧化苦参碱通过抑制Siha细胞增殖和诱导Siha细胞凋亡发挥抗肿瘤作用。

Objective:To explore the effect of oxymatrine on the proliferation and apoptosis of cervical cancer Siha cells infected by human papilloma virus(HPV)16.Methods:Human cervical squamous cell carcinoma Siha cells were used as an in vitro experimental model of cervical cancer infected by HPV16,routinely resuscitated,cultured and passaged;added with different mass concentrations of oxymatrine(0.2 g·L^-1,0.4 g·L^-1,0.8 g·L^-1,1.6 g·L^-1)as the experimental group,and added an equal volume of RPMI-1640 as a solvent control group for 24 hours,48 hours and 72 hours;thiazole blue method was used to detect the changes in cell proliferation activity,flow cytometry was used to analyze the rate of cell apoptosis,and western blotting was used to analyze the expression of apoptosis-related proteins Bcl-2,Bax,and P53.Results:The results of thiazolyl blue showed that the inhibitory effect of oxymatrine on the proliferation of cervical cancer Siha cells infected by HPV16 was mass concentration and time dependent(P<0.05).The results of flow cytometry showed that the apoptosis rate of oxymatrine treated with different concentrations of oxymatrine for 48 hours was compared with that of the control group,and the difference was statistically significant(P<0.05);after 48 hours of oxymatrine treatment,compared with the control group,the ratio of G0/G1 phase of Siha cells in the experimental group increased(P<0.05),the ratio of S phase and G2/M phase decreased(P<0.05),and the cell growth cycle stopped at G1 period.Western blot detection results showed that the expression of apoptosis-related protein Bcl-2 was down-regula-ted,Bax and P53 protein were up-regulated,and the up-regulation(down-regulation)of the experimental group was in a mass concentration-dependent manner.Conclusion:Oxymatrine exerts anti-tumor effects by inhibiting the proliferation of Siha cells and inducing Siha cell apoptosis.