目的:研究异常黑胆质成熟剂总酚与顺铂联合对体外培养的人宫颈癌细胞SiHa增殖的影响,并探讨对凋亡抑制基因Bcl-2和Survivin的表达的影响。方法:异常黑胆质成熟剂总酚分别与顺铂联合作用于SiHa细胞后,荧光倒置显微镜下观察细胞生长情况,流式细胞术检测Siha细胞各组早期凋亡率的变化及Bcl-2的表达,实时荧光定量PCR法检测Survivin的表达。结果:①倒置显微镜下观察单药和联合用药组细胞体积变小变圆,细胞核固缩,核质比减小,胞质浑浊,可见空泡状细胞,部分细胞核膜消失,可见较多脱落圆形死亡细胞。②异常黑胆质成熟剂总酚和顺铂对Siha细胞作用48 h后,流式细胞术检测Siha细胞各组早期凋亡率分别为(14.2±4.06)%和(16.2±1.66)%,明显高于对照组(5.43±0.31)%,差异有统计学意义(P〈0.001)都有很好地促进凋亡的作用,异常黑胆质成熟剂总酚与顺铂联合有交互作用,其促进凋亡的作用比单药更显著(P〈0.01)。③异常黑胆质成熟剂总酚和顺铂单药及总酚与顺铂联合均能降低Bcl-2的表达,并且单药与对照组、联合用药组与单药组之间均有统计学差异(P〈0.001)。④实时荧光定量PCR法检测Survivin的表达定量在异常黑胆质成熟剂总酚组和顺铂组分别为(0.334±0.038)及(0.562±0.035),明显低于对照组Survivin的表达(3.470±0.091),差异有统计学意义(P〈0.001),药物联合组有很好的交互作用,差异有统计学意义(P〈0.001)。结论:异常黑胆质成熟剂总酚与顺铂联合能显著增加SiHa细胞的早期凋亡率,其机制与降低Bcl-2和Survivin表达有关。
Objective:To investigate the effects of total phenolics from abnormal savda munziq combined with cisplatin on Siha cell line of cervical cancer cultured in vitro by detecting express Bcl-2 and Survivin.Methods:Siha cells were cultured in vitro and treated with different drug concentrations;the cisplatins,the total phenolics,the total phenolics combined with cisplatins.Microscope was used to observe the growth of Siha cells.And flow cytometry was used to examine cell apoptosis rate and Bcl-2,real time-PCR was used to detect Survivin expressions.Results:①Siha cells showed Morphologic changes obviously in experimental groups.②The 48h’s apoptosis rates of total phenolics and cisplatin were increased,they were(14.2±4.06)% and(16.2±1.66)% respectively,obviously higher than that control group(5.43±0.31)%,The combinations had interactions effect(P〈0.01) The 48h’s Bcl-2 rates of total phenolics and cisplatin were reduced obviously(P〈0.001).③the total phenolics combined with cisplatin had interactions effect.④The 48h’s Survivin expressions of total phenolics and cisplatin decreased obviously,they were(0.334±0.038)and(0.562±0.035),obviously lower than that control group(3.470±0.091)(P〈0.001),the combinations had the interactions effect(P〈0.001).Conclusions:The total phenolics from abnormal savda munziq can increase apoptosis rate of Siha cells,its mechanism may have relations with decreasing the expressions of Bcl-2 and Survivin.