ATP1A2通过Src/PI3K/Akt信号通路抑制人乳腺癌细胞的侵袭和迁移

Overexpression of α2 subunit of Na-K-ATPase suppresses invasion and migration of human breast cancer cells in vitro by inhibiting the Src/PI3K/Akt signaling pathway

目的探讨ATP1A2在人乳腺癌组织与细胞中的表达水平及其对乳腺癌细胞侵袭和迁移能力的影响。方法分析the Cancer Genome Atlas (TCGA)数据库中ATP1A2在正常乳腺组织和乳腺癌组织中的表达差异,以及在有无远处转移的患者组织中的表达情况;对过表达ATP1A2的MDA-MB-231和SK-BR-3细胞进行细胞划痕实验、Transwell侵袭和迁移实验,以研究其对乳腺癌细胞侵袭和迁移能力的影响;通过Western blot检测Src、PI3K、Akt蛋白的激活情况。结果 ATP1A2在乳腺癌组织中的表达低于正常乳腺组织(P<0.05);远处转移的患者组织中ATP1A2的表达低于无远处转移的患者组织(P<0.05);划痕实验和Transwell侵袭、迁移实验中,ATP1A2过表达组的划痕愈合能力及侵袭和迁移出小室的细胞数均少于对照组(P<0.05);Western blot实验发现:过表达组的Src、PI3K、Akt蛋白的磷酸化水平受到抑制(P<0.05)。结论 ATP1A2在人乳腺癌组织中低表达,其表达可能与乳腺癌患者的远处转移相关;过表达ATP1A2后能显著抑制乳腺癌细胞的侵袭和迁移能力,此种作用可能是通过抑制Src/PI3K/Akt信号通路而产生的。

Objective To investigate the expression of α2 subunit of Na-K-ATPase(ATP1 A2) in human breast cancer and the effects of ATP1 A2 overexpression on the invasion and migration of breast cancer cells in vitro. Methods Based on the data from the Cancer Genome Atlas(TCGA) database, the differences of ATP1 A2 expression were analyzed between normal breast tissue and breast cancer tissue and between breast cancer patients with and without distal metastasis. The breast cancer MDA-MB-231 and SK-BR-3 cells transfected with pEZ-M35-ATP1 A2 plasmid for ATP1 A2 overexpression were evaluated for invasion and migration abilities using wound healing test and Transwell invasion and migration tests;The activation of Src, PI3 K and Akt proteins in the transfected cells was detected with Western blotting. Results The expression of ATP1 A2 was significantly lower in breast cancer tissues than in normal breast tissues, and was also lower in patients with than in those without distal metastasis. In wound healing test and Transwell invasion and migration tests, the breast cancer cells with ATP1 A2 overexpression showed significantly lowered invasion and migration abilities as compared with the control cells. Western blotting showed that ATP1 A2 overexpression significantly reduced the phosphorylation levels of Src, PI3 K and Akt proteins in the breast cancer cells. Conclusion ATP1 A2 is lowly expressed in human breast cancer tissues and shows a likely correlation with distal metastasis. Overexpression of ATP1 A2 can significantly inhibit the invasion and migration of breast cancer cells in vitro possibly by inhibiting the Src/PI3 K/Akt signaling pathway.