NFATc1可能通过调节RhoA/ROCK信号通路影响乳腺癌细胞的增殖与凋亡

目的:检测NFATc1在乳腺癌患者癌及癌旁组织的表达，并探讨其表达对乳腺癌细胞增殖和凋亡的影响及机制。方法:收集于我院就诊的39例乳腺癌患者癌及癌旁组织，采用RT-PCR技术、免疫印迹及免疫组化染色技术检测NFATc1在乳腺癌组织及癌旁组织中的表达状况。利用小干扰RNA（siRNA）构建NFATc1稳定敲除的MCF7及MDA-MB-231细胞系。利用CCK-8试剂盒及克隆形成实验检测NFATc1敲除对乳腺癌细胞增殖的影响，同时利用Ki67染色检测对照组和NFATc1敲除组乳腺癌细胞中Ki67阳性细胞的比例。此外，采用流式细胞学技术检测NFATc1基因敲除对乳腺癌细胞凋亡的影响，最后利用免疫印迹技术分析各组细胞RhoA/ROCK信号通路的表达状况。结果:NFATc1在乳腺癌患者癌组织中的mRNA及蛋白表达显著增高（P＜0.05）。流式细胞学结果表明NFATc1 siRNA干预能够显著促进两组乳腺癌细胞系的凋亡（P＜0.05）。此外，Ki67染色、CCK-8实验及克隆形成实验结果表明NFATc1敲除后两种乳腺癌细胞系的增殖能力明显下降（P＜0.05）。免疫印迹结果揭示，NFATc1敲除能够明显抑制癌细胞中RhoA/ROCK信号通路的激活（P＜0.05）。结论:NFATc1可能通过调控RhoA/ROCK信号通路进而促进乳腺癌细胞增殖，有望成为治疗乳腺癌的新靶点。

Effects of NFATc1 on proliferation and apoptosis of breast cancer cells by regulating RhoA/ROCK signaling pathway

Objective:To detect the expression of NFATc1 in breast cancer and adjacent tissues,and to explore its effect on the proliferation and apoptosis of breast cancer cells.Methods:Adjacent tissues and tumor cancers from thirty-nine breast cancer patients were collected from our hospital.RT-PCR,immunoblotting and immunohistochemical staining were used to detect the expression of NFATc1 in breast cancer tissues and adjacent tissues.Further,the construction of NFATc1 stable knockout MCF7 and MDA-MB-231 using small interfering RNA (siRNA) cell line.The effect of NFATc1 knockdown on the proliferation of breast cancer cells was detected by CCK-8 kit and clone formation assay.The ratio of Ki67 positive cells in breast cancer cells of control group and NFATc1 knockout group was detected by Ki67 staining.In addition,flow cytometry was used to detect the effect of NFATc1 gene knockdown on apoptosis of breast cancer cells.Finally,the expression of RhoA/ROCK signaling pathway in each group was analyzed by immunoblotting.Results:The mRNA and protein expression of NFATc1 in breast cancer patients was significantly increased (P＜0.05).Flow cytometry results showed that NFATc1 siRNA intervention significantly promoted apoptosis in two breast cancer cell lines (P＜0.05).In addition,Ki67 staining,CCK-8 assay and colony formation assay showed that the proliferative capacity of the two breast cancer cell lines was significantly decreased after NFATc1 knockout (P＜0.05).Western blot results revealed that NFATc1 knockdown significantly inhibited the activation of RhoA/ROCK signaling pathway in cancer cells (P＜0.05).Conclusion:Inhibition of NFATc1 can promote the apoptosis of breast cancer cells and inhibit the proliferation of breast cancer cells by inhibiting RhoA/ROCK signaling pathway in breast cancer cells.Therefore,NFATc1 is expected to become a new target for the treatment of breast cancer.