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[摘要]
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 mRNA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。
[Key word]
[Abstract]
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 signifi‐cantly 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.
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[基金项目]
上海市闵行区自然科学研究基金资助项目(No.2016MHZ69)