天津医科大学学报
天津醫科大學學報
천진의과대학학보
Journal of Tianjin Medical University
2015年
6期
466-468,483
,共4页
周岩%宋伟杰%张飞%冀为%田然%牛瑞芳%黎小沛
週巖%宋偉傑%張飛%冀為%田然%牛瑞芳%黎小沛
주암%송위걸%장비%기위%전연%우서방%려소패
人附睾蛋白4%乳腺癌%增殖%凋亡
人附睪蛋白4%乳腺癌%增殖%凋亡
인부고단백4%유선암%증식%조망
human epididymis protein 4%breast cancer%proliferation%apoptosis
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-qPCR)检测15例原发乳腺癌组织及其配对的癌旁正常组织中HE4 mRNA的表达水平。 RT-qPCR和Western blot检测常见乳腺(癌)细胞系中HE4 mRNA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 mRNA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。
目的:探討人附睪蛋白4(HE4)在乳腺癌髮生髮展中的機製研究。方法:採用實時熒光定量PCR(RT-qPCR)檢測15例原髮乳腺癌組織及其配對的癌徬正常組織中HE4 mRNA的錶達水平。 RT-qPCR和Western blot檢測常見乳腺(癌)細胞繫中HE4 mRNA和蛋白錶達水平。通過RNA榦擾技術沉默HE4高錶達的乳腺癌SKBR3細胞中HE4錶達,通過CCK8和剋隆形成實驗分析沉默HE4錶達對SKBR3細胞增殖的影響。流式細胞術檢測沉默HE4錶達對SKBR3細胞凋亡的影響。 Western blot檢測沉默HE4錶達對Erk和Akt燐痠化水平的影響。結果:HE4 mRNA在乳腺癌組織中高錶達;沉默HE4錶達抑製乳腺癌細胞SKBR3細胞增殖能力併促進其凋亡,Erk和Akt蛋白燐痠化水平降低。結論:HE4通過影響Erk和Akt信號通路促進乳腺癌髮生髮展。
목적:탐토인부고단백4(HE4)재유선암발생발전중적궤제연구。방법:채용실시형광정량PCR(RT-qPCR)검측15례원발유선암조직급기배대적암방정상조직중HE4 mRNA적표체수평。 RT-qPCR화Western blot검측상견유선(암)세포계중HE4 mRNA화단백표체수평。통과RNA간우기술침묵HE4고표체적유선암SKBR3세포중HE4표체,통과CCK8화극륭형성실험분석침묵HE4표체대SKBR3세포증식적영향。류식세포술검측침묵HE4표체대SKBR3세포조망적영향。 Western blot검측침묵HE4표체대Erk화Akt린산화수평적영향。결과:HE4 mRNA재유선암조직중고표체;침묵HE4표체억제유선암세포SKBR3세포증식능력병촉진기조망,Erk화Akt단백린산화수평강저。결론:HE4통과영향Erk화Akt신호통로촉진유선암발생발전。
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.