本文選題：miR-224 + 宮頸癌��； 參考：《山西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:驗證miR-224在宮頸癌HeLa細(xì)胞中表達(dá)上調(diào),并探討miR-224對宮頸癌HeLa細(xì)胞中自噬相關(guān)蛋白p62的影響。方法:懫用實時熒光定量PCR法檢測宮頸癌細(xì)胞(HeLa)和正常上皮細(xì)胞(293T)中miR-224的表達(dá)水平,并通過Western Blot技術(shù)檢測上述細(xì)胞中p62蛋白的表達(dá)水平。利用細(xì)胞瞬時轉(zhuǎn)染技術(shù)升高或降低HeLa細(xì)胞中miR-224的表達(dá)水平,設(shè)為miR-224模擬物組(M組)、miR-224模擬物陰性對照組(NC組)、miR-224抑制物組(IN組)、miR-224抑制物陰性對照組(INC組),采用實時熒光定量PCR方法確定轉(zhuǎn)染效率。確定轉(zhuǎn)染成功后利用Western blot方法檢測各組細(xì)胞中自噬相關(guān)基因蛋白p62的表達(dá)水平。結(jié)果:1、HeLa細(xì)胞中miR-224的表達(dá)水平(0.151±0.014)高于293T細(xì)胞中miR-224的表達(dá)水平(6.732±1.33),差異有統(tǒng)計學(xué)意義(P0.05);2、HeLa細(xì)胞中p62蛋白的表達(dá)水平(0.608±0.021)低于293T細(xì)胞中p62蛋白的表達(dá)水平(0.888±0.033),差異有統(tǒng)計學(xué)意義(P0.05);3、HeLa細(xì)胞轉(zhuǎn)染后,miR-224模擬物組(M組)中miR-224的表達(dá)量較miR-224模擬物陰性對照組(NC組)增加了25.13倍,其p62蛋白表達(dá)水平降低至74%;與miR-224抑制物陰性對照組(INC組)相比,miR-224抑制物組(IN組)中miR-224表達(dá)降低至24%,而p62蛋白表達(dá)水平增加了1.71倍,差異有統(tǒng)計學(xué)意義(P0.05)。結(jié)論:1、宮頸癌HeLa細(xì)胞中miR-224呈高表達(dá),提示miR-224在宮頸癌中可能發(fā)揮了促癌作用;2、宮頸癌HeLa細(xì)胞自噬活性高于正常上皮細(xì)胞;過表達(dá)miR-224可能會促進(jìn)宮頸癌HeLa細(xì)胞的自噬活性;低表達(dá)miR-224可能會抑制宮頸癌HeLa細(xì)胞的自噬活性,提示miR-224可能是通過影響細(xì)胞的姿勢進(jìn)而影響宮頸癌細(xì)胞的生物學(xué)行為。
[Abstract]:Aim: to investigate the upregulation of miR-224 expression in cervical cancer HeLa cells and the effect of miR-224 on autophagy associated protein p62 in cervical cancer HeLa cells. Methods: the expression of miR-224 in cervical carcinoma cell line HeLa) and normal epithelial cells was detected by real-time fluorescence quantitative PCR, and the expression of p62 protein was detected by Western Blot technique. The transient transfection technique was used to increase or decrease the expression of miR-224 in HeLa cells. The transfection efficiency was determined by real-time fluorescence quantitative PCR (real-time fluorescence quantitative PCR) method in the miR-224 mimics group and M group as the negative control group and NC group as the control group and in the IN group as the negative control group. The transfection efficiency was determined by real-time fluorescence quantitative PCR. The expression of autophagy associated gene protein p62 was detected by Western blot method after transfection. Results the expression level of miR-224 in HeLa cells was 0.151 鹵0.014), which was higher than that in 293T cells (6.732 鹵1.33). There was a significant difference in the expression level of p62 protein (0.608 鹵0.021) in HeLa cells (P 0.05) and in 293T cells (0.888 鹵0.033). After transfection, the expression of miR-224 was 25.13 times higher than that of miR-224 negative control group (M group). Compared with the control group, the expression of p62 protein in the control group was significantly lower than that in the control group (n = 24), but the expression level of p62 protein was 1.71 times higher than that in the control group (P 0.05, P 0.05), and the expression level of p62 protein in the control group was significantly higher than that in the control group (P < 0.05), but the expression of p62 protein in the control group was significantly higher than that in the control group (P < 0.05). Conclusion the overexpression of miR-224 in cervical cancer HeLa cells suggests that miR-224 may play a role in promoting cancer in cervical cancer, and the autophagy activity of HeLa cells in cervical cancer is higher than that of normal epithelial cells, and overexpression of miR-224 may promote the autophagy activity of HeLa cells in cervical carcinoma. The low expression of miR-224 may inhibit the autophagy activity of cervical cancer HeLa cells, suggesting that miR-224 may affect the biological behavior of cervical cancer cells by affecting the posture of the cells.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R737.33

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