人附睾蛋白4在乳腺癌发生发展中的机制研究

Action mechanism of human epididymis protein 4 in development and progression of breast cancer

目的:探讨人附睾蛋白4(HE4)在乳腺癌发生发展中的机制研究。方法:采用实时荧光定量PCR(RT-q PCR)检测15例原发乳腺癌组织及其配对的癌旁正常组织中HE4 m RNA的表达水平。RT-q PCR和Western blot检测常见乳腺(癌)细胞系中HE4 m RNA和蛋白表达水平。通过RNA干扰技术沉默HE4高表达的乳腺癌SKBR3细胞中HE4表达,通过CCK8和克隆形成实验分析沉默HE4表达对SKBR3细胞增殖的影响。流式细胞术检测沉默HE4表达对SKBR3细胞凋亡的影响。Western blot检测沉默HE4表达对Erk和Akt磷酸化水平的影响。结果:HE4 m RNA在乳腺癌组织中高表达;沉默HE4表达抑制乳腺癌细胞SKBR3细胞增殖能力并促进其凋亡,Erk和Akt蛋白磷酸化水平降低。结论:HE4通过影响Erk和Akt信号通路促进乳腺癌发生发展。

Objective：To explore the mechanism of human epididymis protein 4 （HE4） in development and progression of breast cancer. Methods：Reverse transcription quantitative PCR （RT-qPCR） was used to detect the HE4 mRNA expression levels in primary breast cancer tissues and the paired adjacent normal tissues. The HE4 mRNA and protein expression was also detected in breast epithelial cell lines by RT-qPCR and western blot, respectively. The RNAi was used to knock down the HE4 expression in HE4 high expression level cell line SKBR3, CCK8 and colony formation assays were performed to analyze the proliferation ability. Flow cytometry was applied to detect the apoptotic cells in HE4-depleted cells. The phosphorylation expression levels of Erk and Akt was evaluated by western blot. Results：The HE4 mRNA was up-regulated in primary breast cancer tissues compared with the adjacent normal breast tissues. Depletion of HE4 expression suppressed the proliferation and promoted the apoptosis in SKBR3 breast cancer cell line. Furthermore, the phosphorylation expression levels of Erk and Akt were reduced in HE4-depleted SKBR3 cells. Conclusion：HE4 promotes breast cancer development and progression through Erk and Akt signaling.