TP53 G199X和V157fs点突变对卵巢癌A2780细胞功能的影响

doi:10.13481/j.1671-587x.20200214

摘要/Abstract

摘要： 目的：通过细胞实验对致病候选基因TP53进行功能实验，探讨TP53G199X和V157fs点突变对卵巢癌A2780细胞蛋白表达、增殖、迁移和侵袭的影响。方法：选择无内源性TP53基因表达的卵巢癌A2780细胞，分为野生型组、G199X组、V157fs组和空载体组。构建TP53野生型质粒，利用点突变试剂盒构建筛选得到的G199X和V157fs 2个点突变的质粒，利用脂质体转染法在A2780卵巢癌细胞中转染野生型、突变型和空载体相关质粒。采用Western blotting法检测各组A2780细胞中P53蛋白表达，CCK-8实验法检测各组A2780细胞增殖活性，划痕实验检测各组A2780细胞划痕愈合率，Transwell细胞侵袭实验检测各组A2780细胞的侵袭细胞数。结果：A2780细胞经转染后，TP53 G199X和V157fs突变型组及空载体组细胞中无P53蛋白表达。与野生型组比较，G199X组、V157fs组空载体组细胞增殖活性明显升高（P<0.01）；G199X组和V157fs组A2780细胞划痕愈合率升高（P<0.05或P<0.01）；G199X组、V157fs组和空载体组侵袭细胞数明显升高（P<0.01）。结论：TP53G199X和V157fs 2个点突变在A2780细胞中可终止P53蛋白的表达并促进卵巢癌A2780细胞的增殖、迁移和侵袭。

关键词: TP53基因, 点突变, 细胞增殖, 细胞迁移, 细胞侵袭

Abstract: Objective: To perform the function experiment in the pathogenic candidate gene TP53 via cell experiment,and to investigate the effects of TP53 G199X and V157fs point mutations on the protein expression, proliferation, migration and invasion of the ovarian cancer A2780 cells. Methods: The ovarian cancer A2780 cells without endogenous TP53 gene expression were selected and divided into wild-type group, G199X group, V157fs group and empty vector group. The TP53 wild-type plasmid was constructed, and the two point mutation plasmids of G199X and V157fs were constructed by using the point mutation kit. The wild-type, mutant and empty vector plasmids were transfected into the A2780 cells by lipofection. Western blotting method was used to detect the the expressions of P53 protein in the A2780 cells in various goups. The proliferation activities of A2780 cells in various groups were detected by CCK-8 assay. The rates of scratch healing of A2780 cells in various groups were detected by scratch assay. Transwell assay was used to examine the number of invasion cells of A2780 cells in various groups. Results: After transfection of A2780 cells, the P53 protein didn't express in the cells in G199X group, V157fs group and empty vector group. Compared with wild-type group, the proliferation activities of the cells in G199X group,V157fs group and empty vector group were significantly increased(P<0.01);the scratch healing rates of the cells in G199X group and V157fs group were increased (P<0.05 or P<0.01); the number of invasion cells in G199X group,V157fs group and empty vector group was increased(P<0.01). Conclusion: Two point mutations of TP53 G199X and V157fs can stop the expression of P53 protein in the A2780 cells and promote the proliferation, migration and invasion of ovarian cancer A2780 cells.

Key words: TP53 gene, point mutation, cell proliferation, cell migration, cell invasion

中图分类号:

R737.31

聂蔓, 岳军. TP53 G199X和V157fs点突变对卵巢癌A2780细胞功能的影响[J]. 吉林大学学报(医学版), 2020, 46(02): 292-296.

NIE Man, YUE Jun. Effects of TP53 G199X and V157fs on function of ovarian cancer A2780 cells[J]. Journal of Jilin University(Medicine Edition), 2020, 46(02): 292-296.

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