【摘要】：目的自噬是細胞在不良環(huán)境中自我吞噬的生物學現(xiàn)象,在惡性腫瘤發(fā)病機制中具有重要作用,缺氧是誘導細胞自噬的重要因素。本研究探討不同氧環(huán)境中卵巢癌HO-8910細胞let-7a的表達變化及對缺氧細胞自噬、增殖的影響。方法實驗分常氧組、缺氧組、缺氧+miR-NC-mimics組和缺氧+let-7a-mimics組4組,各組細胞經(jīng)處理48h后:吖啶橙染色觀察細胞自噬發(fā)生情況;MTT法分析細胞增殖變化,流式細胞術(shù)檢測細胞周期分布;Real time PCR分析let-7a、Beclin-1和LC3-Ⅱ的表達變化,蛋白質(zhì)印跡試驗分析Beclin-1和LC3-Ⅱ蛋白表達變化。結(jié)果 HO-8910細胞經(jīng)缺氧處理后自噬率顯著增高至(34.46±5.13)%,F=49.932,P=0.002;在缺氧基礎(chǔ)上轉(zhuǎn)染miR-NC-mimics后,自噬率變化不大,F=0.110,P=0.756;而轉(zhuǎn)染let-7a-mimics后,細胞自噬率顯著降低至(4.56±2.04)%,F=68.186,P0.001。缺氧使G0/G1期細胞數(shù)量明顯增至(70.43±3.19)%,F=23.216,P=0.008;同時轉(zhuǎn)染miR-NC-mimics后,其數(shù)量變化差異無統(tǒng)計學意義,F=0.467,P=0.532;而在缺氧條件下轉(zhuǎn)染let-7a-mimics,其數(shù)量則增至(82.37±3.45)%,F=67.423,P=0.001;并伴隨細胞增殖抑制率顯著上升,F=24.602,P=0.007。缺氧條件下以及缺氧同時轉(zhuǎn)染miR-NC-mimics后,let-7a表達下調(diào),而Beclin-1、LC3-Ⅱ表達上調(diào);而在缺氧基礎(chǔ)上轉(zhuǎn)染let-7a-mimics后,let-7a表達明顯增強,F=295.623,P0.001;Beclin-1蛋白表達下調(diào),F=33.285,P=0.002;LC3-Ⅱ也明顯下調(diào),F=41.969,P=0.001。結(jié)論卵巢癌HO-8910細胞在缺氧狀態(tài)時,let-7a表達下調(diào),細胞啟動適度自噬保護機制,介導細胞逃避缺氧環(huán)境使細胞繼續(xù)快速生長,當上調(diào)let-7a表達后,缺氧所致的適度自噬保護作用被消減,細胞增殖明顯受抑。
[Abstract]:Objective autophagy is a biological phenomenon of self-phagocytosis of cells in a bad environment and plays an important role in the pathogenesis of malignant tumors. Hypoxia is an important factor in inducing autophagy. The purpose of this study was to investigate the changes of let-7a expression in ovarian cancer HO-8910 cells and its effects on the autophagy and proliferation of anoxic cells in different oxygen environments. Methods the cells were divided into four groups: normoxic group, hypoxic miR-NC-mimics group and hypoxic let-7a-mimics group. After 48 hours of treatment, the autophagy was observed by acridine orange staining. The expression of let-7a,Beclin-1 and LC3- 鈪，

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