Abstract

The ovarian cancer cell line A2780 without endogenous TP53 expression was selected. A plasmid carrying wild-type TP53, TP53 with two point mutations G199X and V157fs, or an empty vector was transfected in A2780 cells with lipofection. Western blot analysis was used to investigate whether the mutant plasmid affects the expression of P53 protein. In addition, the effect of mutation on the migration of A2780 cells was detected with scratch assay, while that on cell invasion was evaluated with transwell assay. We also studied the effect of mutation on the proliferation of A2780 cells with the cell counting kit-8 (CCK-8) assay. As a result, we found that the cells transfected with TP53 G199X and V157fs mutants and empty vector failed to express P53 protein. The migration ability of the cells expressing the two mutant genes was higher than that of the cells expressing the wild-type gene. The invasion ability of the two mutant groups was significantly higher than that of the wild-type group. Cell proliferation was also higher for the two mutant groups and the empty vector group than for the wild-type group. Thus, the two point mutations in TP53 (G199X and V157fs) gene could terminate the expression of P53 protein and promote migration, invasion, and proliferation of ovarian cancer A2780 cells.

中文翻译：

---

TP53中的两个突变促进人类卵巢癌A2780细胞的迁移，侵袭和增殖

摘要

选择没有内源性TP53表达的卵巢癌细胞系A2780 。携带野生型的质粒TP53，TP53与两个点突变G199X和V157fs，或空载体在A2780细胞与脂转染转染。使用蛋白质印迹分析来研究突变质粒是否影响P53蛋白的表达。另外，通过刮擦测定法检测突变对A2780细胞迁移的影响，而通过transwell测定法评价对细胞侵袭的影响。我们还使用细胞计数试剂盒8（CCK-8）分析研究了突变对A2780细胞增殖的影响。结果，我们发现用TP53转染的细胞G199X和V157fs突变体和空载体未能表达P53蛋白。表达两个突变基因的细胞的迁移能力高于表达野生型基因的细胞的迁移能力。两个突变组的侵袭能力明显高于野生型组。两个突变组和空载体组的细胞增殖也高于野生型组。因此，TP53基因的两个点突变（G199X和V157fs）可以终止P53蛋白的表达并促进卵巢癌A2780细胞的迁移，侵袭和增殖。