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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 effect of the combination of total phenols and cisplatin on the proliferation of human cervical cancer cell SiHa cultured in vitro and to explore the effect on the expression of apoptosis-suppressing genes Bcl-2 and Survivin. Methods: Total phenols of abnormal savda were combined with cisplatin respectively in SiHa cells. Fluorescence inverted microscope was used to observe the growth of SiH cells. Flow cytometry was used to detect the changes of early apoptotic rate of Siha cells and the expression of Bcl-2 Expression of Survivin by real-time fluorescence quantitative PCR. Results: (1) Under inverted microscope, the volume of cells in single drug group and combined drug group became smaller and round, the nuclei were pyknotic, the ratio of nucleus to cytoplasm was decreased, the cytoplasm was cloudy, vacuolar cells were visible, and some nuclear membrane disappeared. Dead cells. (2) The apoptosis rates of Siha cells were (14.2 ± 4.06)% and (16.2 ± 1.66)%, respectively, which were significantly higher than those of Siha cells treated with total phenols and cisplatin of abnormal savda Higher than the control group (5.43 ± 0.31)%, the difference was statistically significant (P <0.001) have a good role in promoting apoptosis, Abnormal Savda saponin total phenol and cisplatin combined interaction, and its promotion Apoptosis was more significant than single drug (P <0.01). ③ Abnormal savda mature agent total phenols and cisplatin alone and total phenol and cisplatin can reduce the expression of Bcl-2, and the single drug and the control group, the combination group and the single drug group were statistically Difference (P <0.001). ④ The expression of Survivin was detected by real-time fluorescence quantitative PCR in the total phenolic group and cisplatin group (0.334 ± 0.038) and (0.562 ± 0.035), which was significantly lower than that in the control group (3.470 ± 0.091), the difference was statistically significant (P <0.001), the drug combination group had a good interaction, the difference was statistically significant (P <0.001). CONCLUSION: The combination of total phenol and cisplatin in abnormal savda mature maturity can significantly increase the early apoptosis rate of SiHa cells, which is related to the decrease of Bcl-2 and Survivin expression.