自噬对金雀异黄酮抑制宫颈癌细胞增殖的影响

发布时间：2018-11-19 08:25

【摘要】：目的金雀异黄酮对多种肿瘤有抑制作用。文中研究金雀异黄酮对宫颈癌Hela细胞增殖的抑制作用和对自噬的诱导,以及自噬在此过程中的作用机制。方法体外培养人宫颈癌Hela细胞,将其分为对照组、金雀异黄酮组和3-甲基腺嘌呤(3-MA)+金雀异黄酮组,对照组只加培养液,金雀异黄酮组用(25、50、100μmol/L)金雀异黄酮处理,3-MA+金雀异黄酮组先用5 mmol/L 3-MA预处理1 h后加入100μmol/L金雀异黄酮,采用MTT法检测各组细胞增殖抑制率的变化;透射电镜观察细胞自噬超微结构的改变;Western blot检测自噬相关蛋白P62及Beclin-1的表达情况;免疫荧光法检测LC3A/B的表达,分析金雀异黄酮对细胞自噬水平的影响。结果与对照组Hela细胞增殖抑制率(0)相比,25、50、100μmol/L金雀异黄酮处理后,Hela细胞增殖的抑制率(20.9%±1.3%,33.5%±1.6%,46.5%±3.2%)逐渐增高(P0.01)。使用不同浓度(25、50、100μmol/L)金雀异黄酮处理Hela细胞48 h后,随着金雀异黄酮浓度增高Beclin-1表达逐渐增强,P62的表达逐渐降低。激光共聚焦显微镜下观察可见,100μmol/L金雀异黄酮处理组He La细胞细胞质中可见到大量LC3A/B绿色荧光分布,且荧光强度较强。金雀异黄酮(100μmol/L)处理48 h后细胞质中可见大量空泡状结构及包含部分细胞质成分形成双层膜结构的自噬体。金雀异黄酮组Hela细胞的增殖抑制率较3-MA+金雀异黄酮组低[(46.5±3.2)%vs(58.2±2.2)%,P0.01]。结论金雀异黄酮可抑制Hela细胞增殖,并诱导自噬增强。
[Abstract]:Objective isoflavone has inhibitory effect on many kinds of tumors. The inhibitory effect of genistein on the proliferation of cervical cancer Hela cells and the induction of autophagy and the mechanism of autophagy in this process were studied. Methods Human cervical cancer Hela cells were cultured in vitro and divided into control group, genistein group and 3-methyladenine (3-MA) genistein group. The genistein group was treated with (25 5 0100 渭 mol/L) genistein, and the 3-MA genistein group was pretreated with 5 mmol/L 3-MA for 1 h, then 100 渭 mol/L genistein was added. The inhibition rate of cell proliferation in each group was detected by MTT assay. Ultrastructural changes of autophagy were observed by transmission electron microscope (TEM). The expression of autophagy associated protein P62 and Beclin-1 was detected by; Western blot, and the expression of LC3A/B was detected by immunofluorescence assay, and the effect of genistein on autophagy level was analyzed. Results compared with the control group, the inhibitory rate of Hela cell proliferation was (20.9% 卤1.3% 卤1.650%) after treatment with 2550100 渭 mol/L genistein. 46.5% 卤3.2%) (P0.01). After treatment with different concentrations (25550100 渭 mol/L) of genistein for 48 h, the expression of Beclin-1 increased gradually and the expression of P62 decreased with the concentration of genistein increased. Laser confocal microscopy showed that a large number of LC3A/B green fluorescence could be observed in the cytoplasm of He La treated with 100 渭 mol/L genistein, and the fluorescence intensity was strong. After 48 h treatment with genistein (100 渭 mol/L), a large number of vacuolated structures and autophagy with bilayer membrane structure were found in the cytoplasm. The inhibitory rate of Hela cell proliferation in genistein group was lower than that in 3-MA genistein group [(46.5 卤3.2)% vs (58.2 卤2.2)%, P0.01]. Conclusion genistein can inhibit the proliferation of Hela cells and induce the enhancement of autophagy.
【作者单位】： 哈尔滨医科大学附属肿瘤医院妇二科;哈尔滨医科大学肿瘤研究所;哈尔滨红十字中心医院;
【基金】：黑龙江省自然科学基金(D201136) 黑龙江省政府博士后启动基金(LRB06-267)
【分类号】：R737.33
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