MiR-224对宫颈癌HeLa细胞自噬相关蛋白p62的影响

发布时间：2018-05-17 05:30

  本文选题：miR-224 + 宫颈癌　； 参考：《山西医科大学》2017年硕士论文

【摘要】：目的:验证miR-224在宫颈癌HeLa细胞中表达上调,并探讨miR-224对宫颈癌HeLa细胞中自噬相关蛋白p62的影响。方法:懫用实时荧光定量PCR法检测宫颈癌细胞(HeLa)和正常上皮细胞(293T)中miR-224的表达水平,并通过Western Blot技术检测上述细胞中p62蛋白的表达水平。利用细胞瞬时转染技术升高或降低HeLa细胞中miR-224的表达水平,设为miR-224模拟物组(M组)、miR-224模拟物阴性对照组(NC组)、miR-224抑制物组(IN组)、miR-224抑制物阴性对照组(INC组),采用实时荧光定量PCR方法确定转染效率。确定转染成功后利用Western blot方法检测各组细胞中自噬相关基因蛋白p62的表达水平。结果:1、HeLa细胞中miR-224的表达水平(0.151±0.014)高于293T细胞中miR-224的表达水平(6.732±1.33),差异有统计学意义(P0.05);2、HeLa细胞中p62蛋白的表达水平(0.608±0.021)低于293T细胞中p62蛋白的表达水平(0.888±0.033),差异有统计学意义(P0.05);3、HeLa细胞转染后,miR-224模拟物组(M组)中miR-224的表达量较miR-224模拟物阴性对照组(NC组)增加了25.13倍,其p62蛋白表达水平降低至74%;与miR-224抑制物阴性对照组(INC组)相比,miR-224抑制物组(IN组)中miR-224表达降低至24%,而p62蛋白表达水平增加了1.71倍,差异有统计学意义(P0.05)。结论:1、宫颈癌HeLa细胞中miR-224呈高表达,提示miR-224在宫颈癌中可能发挥了促癌作用;2、宫颈癌HeLa细胞自噬活性高于正常上皮细胞;过表达miR-224可能会促进宫颈癌HeLa细胞的自噬活性;低表达miR-224可能会抑制宫颈癌HeLa细胞的自噬活性,提示miR-224可能是通过影响细胞的姿势进而影响宫颈癌细胞的生物学行为。
[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 卤0.014), which was higher than that in 293T cells (6.732 卤1.33). There was a significant difference in the expression level of p62 protein (0.608 卤0.021) in HeLa cells (P 0.05) and in 293T cells (0.888 卤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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.33

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