摘要
目的:探讨微小RNA-200a(miR-200a)表达改变对乳腺癌细胞恶性生物学行为的影响及其调节机制。方法:RT-qPCR法检测人乳腺癌细胞株MDA-MB-231、MDA-MB-468和MCF-7中miR-200a的表达水平与正常人乳腺上皮细胞株MCF-10A的差异。CCK-8法检测转染miR-200a mimic和miR-inhibitor后MDA-MB-231细胞活力的变化。采用流式细胞术及Transwell实验检测转染后MDA-MB-231细胞凋亡及侵袭能力的变化。采用RT-qPCR和Western blot检测上皮钙黏素(E-cadherin)、神经钙黏素(N-cadherin)、SIP1、Snail、Twist、ZEB1和ZEB2 mRNA及蛋白表达的变化。结果:与MCF-10A细胞相比,miR-200a在MDA-MB-231细胞中表达显著降低(P<0.05)。过表达miR-200a使MDA-MB-231细胞活力降低(P<0.05),凋亡率升高(P<0.05),侵袭能力下降(P<0.05),SIP1、N-cadherin、Snail、Twist、ZEB1和ZEB2的mRNA及蛋白表达显著下调(P<0.05),而E-cadherin的mRNA及蛋白表达显著增加(P<0.05);抑制miR-200a的表达使上述结果逆转(P<0.05)。结论:上调miR-200a能够抑制MDA-MB-231细胞的活力和侵袭,促进MDA-MB-231细胞的凋亡。miR-200a可能通过抑制上皮-间充质转化调控乳腺癌的恶性生物学行为。
AIM:To investigate the effect of microRNA-200a(miR-200a)on the malignant biological beha-viors of breast cancer cells and its regulatory mechanism.METHODS:The expression of miR-200a in human breast can-cer cell lines MDA-MB-231,MDA-MB-468 and MCF-7,and normal human mammary epithelial cell line MCF-10A was detected by RT-qPCR.CCK-8 assay was used to detect the viability of MDA-MB-231 cells after transfection with miR-200a mimic or miR-200a inhibitor.Flow cytometry method and Transwell assay were used to detect the apoptosis and invasive ability of MDA-MB-231 cells after transfection with miR-200a mimic or miR-200a inhibitor.The expression of SIP1,E-cadherin,N-cadherin,Snail,Twist,ZEB1 and ZEB2 at mRNA and protein levels was determined by RT-qPCR and Western blot.RESULTS:Compared with MCF-10A cells,the lowest expression of miR-200a was observed in the MDA-MB-231 cells(P<0.05).Over-expression of miR-200a attenuated the viability of MDA-MB-231 cells(P<0.05),increased apoptosis(P<0.05)and decreased the invasion ability(P<0.05).The expression of SIP1,N-cadherin,Snail,Twist,ZEB1 and ZEB2 at mRNA and protein levels was also significantly down-regulated,while the mRNA and protein expression of E-cadherin was significantly increased(P<0.05).Transfection with miR-200a inhibitor reversed the above results.CONCLUSION:Up-regulation of miR-200a inhibits the viability and invasion ability of MDA-MB-231 cells and promotes the apoptosis of MDA-MB-231 cells.miR-200a may regulate the biological behaviors of breast cancer by inhibiting epithelial-mesenchymal transition.
作者
窦艳
邱鹏
陈江伟
DOU Yan;QIU Peng;CHEN Jiang-wei(Department of Pathology,The People's Hospital of Xingtai city,Xingtai 054000,China;Department of Oncology,The People’s Hospital of Xingtai city,Xingtai 054000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2019年第10期1851-1857,共7页
Chinese Journal of Pathophysiology
基金
邢台市科研基金资助项目(No.2018zc147)
