【摘要】：目的金雀異黃酮對(duì)多種腫瘤有抑制作用。文中研究金雀異黃酮對(duì)宮頸癌Hela細(xì)胞增殖的抑制作用和對(duì)自噬的誘導(dǎo),以及自噬在此過程中的作用機(jī)制。方法體外培養(yǎng)人宮頸癌Hela細(xì)胞,將其分為對(duì)照組、金雀異黃酮組和3-甲基腺嘌呤(3-MA)+金雀異黃酮組,對(duì)照組只加培養(yǎng)液,金雀異黃酮組用(25、50、100μmol/L)金雀異黃酮處理,3-MA+金雀異黃酮組先用5 mmol/L 3-MA預(yù)處理1 h后加入100μmol/L金雀異黃酮,采用MTT法檢測各組細(xì)胞增殖抑制率的變化;透射電鏡觀察細(xì)胞自噬超微結(jié)構(gòu)的改變;Western blot檢測自噬相關(guān)蛋白P62及Beclin-1的表達(dá)情況;免疫熒光法檢測LC3A/B的表達(dá),分析金雀異黃酮對(duì)細(xì)胞自噬水平的影響。結(jié)果與對(duì)照組Hela細(xì)胞增殖抑制率(0)相比,25、50、100μmol/L金雀異黃酮處理后,Hela細(xì)胞增殖的抑制率(20.9%±1.3%,33.5%±1.6%,46.5%±3.2%)逐漸增高(P0.01)。使用不同濃度(25、50、100μmol/L)金雀異黃酮處理Hela細(xì)胞48 h后,隨著金雀異黃酮濃度增高Beclin-1表達(dá)逐漸增強(qiáng),P62的表達(dá)逐漸降低。激光共聚焦顯微鏡下觀察可見,100μmol/L金雀異黃酮處理組He La細(xì)胞細(xì)胞質(zhì)中可見到大量LC3A/B綠色熒光分布,且熒光強(qiáng)度較強(qiáng)。金雀異黃酮(100μmol/L)處理48 h后細(xì)胞質(zhì)中可見大量空泡狀結(jié)構(gòu)及包含部分細(xì)胞質(zhì)成分形成雙層膜結(jié)構(gòu)的自噬體。金雀異黃酮組Hela細(xì)胞的增殖抑制率較3-MA+金雀異黃酮組低[(46.5±3.2)%vs(58.2±2.2)%,P0.01]。結(jié)論金雀異黃酮可抑制Hela細(xì)胞增殖,并誘導(dǎo)自噬增強(qiáng)。
[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% 鹵1.3% 鹵1.650%) after treatment with 2550100 渭 mol/L genistein. 46.5% 鹵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 鹵3.2)% vs (58.2 鹵2.2)%, P0.01]. Conclusion genistein can inhibit the proliferation of Hela cells and induce the enhancement of autophagy.
【作者單位】： 哈爾濱醫(yī)科大學(xué)附屬腫瘤醫(yī)院婦二科;哈爾濱醫(yī)科大學(xué)腫瘤研究所;哈爾濱紅十字中心醫(yī)院;
【基金】：黑龍江省自然科學(xué)基金(D201136) 黑龍江省政府博士后啟動(dòng)基金(LRB06-267)
【分類號(hào)】：R737.33
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本文編號(hào)：2341688