Tspan29在乳腺癌组织中的表达及其对MCF-7和MDA-MB-231细胞恶性生物学行为的影响

Expression of Tspan29 in breast cancer tissues and its effect on malignant biological behaviors of MCF-7 and MDA-MB-231 cells

目的:检测4次穿膜蛋白29(tetraspanins-29,Tspan29)在乳腺癌组织和细胞系中的表达水平,探讨敲低Tspan29对乳腺癌MCF-7和MDA-MB-231细胞增殖、迁移、侵袭及上皮间质转化(epithelieal-mesenchymal transition,EMT)的影响。方法:收集2017年6月至2018年2月复旦大学附属肿瘤医院闵行分院手术切除的20例乳腺癌患者的癌组织和相应的癌旁组织标本,以及乳腺癌细胞系MCF-7、MDA-MB-231和人乳腺上皮细胞MDA-kb2,用qPCR和Western blotting检测乳腺癌组织和细胞系中Tspan29的表达水平。通过siTspan29对MCF-7、MDA-MB-231细胞中Tspan29进行干扰,用qPCR法检测转染细胞中Tspan29mRNA和蛋白的表达水平,PCR芯片法检测MCF-7细胞中EMT相关基因的表达,用CCK-8法、Transwell实验检测MCF-7和MDA-MB-231细胞的增殖、迁移和侵袭能力。结果:乳腺癌组织中Tspan29 mRNA和蛋白的表达水平显著高于癌旁组织(均P<0.01),MCF-7和MDA-MB-231细胞中Tspan29 m RNA和蛋白的表达水平显著高于MDA-kb2细胞(均P<0.01)。siTspan29干扰后,MCF-7细胞中Tspan29 mRNA和蛋白的表达水平显著下降(均P<0.05);MCF-7细胞中EMT相关基因中有2个基因显著上调,有7个基因显著下调;MCF-7和MDA-MB-231细胞的增殖、迁移及侵袭能力显著下降(均P<0.05)。结论:Tspan29在乳腺癌组织及细胞系中表达水平显著上调,敲低Tspan29对乳腺癌细胞的增殖、迁移和侵袭有显著的抑制作用。

Objective:To investigate the expression of tetraspanins-29(Tspan29)in breast cancer tissues and cell lines and to explore the effect of Tspan29 knockdown on proliferation,invasion,migration and epithelial-mesenchymal transition(EMT)of breast cancer MCF-7 and MDA-MB-231 cells.Methods:A total of 20 pairs of breast cancer tissues and corresponding para-cancerous tissues resected in Minhang Branch of Cancer Hospital Affiliated to Fudan University from June 2017 to February 2018 were collected for this study;in addition,breast cancer cell lines MCF-7,MDA-MB-231 and human breast epithelial MDA-kb2 cells were also collected.The mRNA and protein expressions of Tspan29 in above mentioned tissues and cell lines were detected by Real-time quantitative(qPCR)and Western blotting.The expression of Tspan29 in MCF-7 and MDA-MB-231 cells was interfered by siRNA.qPCR was used to detect the mRNA and protein expressions of Tspan29.PCR microarray was used to examine the expressions of EMT-related genes in MCF-7 cells.CCK-8 assay and Transwell were used to detect cell proliferation,migration and invasion of MCF-7 and MDA-MB-231 cells.Results:The mRNA and protein expressions of Tspan29 in breast cancer tissues were significantly higher than that in para-cancerous tissues(all P<0.01);and the mRNA and protein expressions of Tspan29 in MCF-7 and MDA-MB-231 cells were significantly higher than that in MDA-kb2 cells(P<0.01).After being interfered with siTspan29,the mRNA and protein expressions of Tspan29 were significantly down-regulated in MCF-7 cells(all P<0.05);the proliferation,invasion and migration of MCF-7 and MDA-MB-231 cells were significantly inhibited(all P<0.05);and among the EMT-related genes,two were significantly up-regulated while 7 were down-regulated.Conclusion:Tspan29 is significantly up-regulated in breast cancer tissues and cell lines,and knockdown of Tspan29 significantly inhibits the proliferation,invasion and migration of breast cancer cells.