摘要
背景与目的:既往研究表明,微小RNA-340(miR-340)能负性调控多种肿瘤的进展,但其在乳腺癌细胞增殖和凋亡中的研究较少,本研究旨在探讨miR-340对乳腺癌MDA-MB231细胞增殖和凋亡的作用。方法:利用脂质体LipofectamineTM2000将pre-miR-340或anti-miR-340瞬时转染至乳腺癌MDA-MB231细胞,通过RT-PCR检测miR-340 mRNA的水平,蛋白质印迹法(Western blot)检测cleaved-caspase-3蛋白的表达,MTT比色法检测细胞增殖的抑制情况,流式细胞仪检测细胞凋亡。结果:Pre-miR-340增加了MDA-MB231细胞中miR-340表达,同时增加cleaved-caspase-3蛋白表达、抑制MDA-MB231细胞增殖并促进其凋亡。而anti-miR-340抑制了MDAMB231细胞中miR-340表达,并且抑制cleaved-caspase-3蛋白表达,促进了MDA-MB231细胞增殖,抑制了MDAMB231细胞的凋亡。结论:miR-340转染后能上调MDA-MB231细胞中cleaved-caspase-3蛋白的表达从而抑制细胞增殖,促进其凋亡。
Background and purpose：MicroRNA-340 （miR-340） has been demonstrated to play a role of negative regulation in many kinds of tumor, however, there are few reports about the relationship between miR-340 in proliferation and apoptosis of breast cancer cell. This study was aimed to explore the effect of miR-340 on proliferation and apoptosis of breast cancer cell MDA-MB231. Methods： The pre-miR-340 or anti-miR-340 were transiently transfected into breast cancer cell MDA-MB231 with Lipofectamine^TM2000. miR-340 level was detected by RT-PCR. The Western blot was performed to detect the protein level of cleaved-caspase-3. The inhibition rate of cell proliferation was evaluated by MTT assay. The cell apoptosis was studied by lfow cytometry. Results：The pre-miR-340 facilitated the expression of miR-340 in MDA-MB231 cells. The pre-miR-340 enhanced the protein level of cleaved-caspase-3, inhibited the proliferation of MDA-MB231 cells and increased its apoptosis. On the contrary, the expression of miR-340 was inhibited by anti-miR-340 in MDA-MB231 cells. The protein level of cleaved-caspase-3 was reduced after the anti-miR-340-transfected MDA-MB231 cells. Anti-miR-340 promoted the proliferation of MDA-MB231 cells. Decreased apoptosis of MDA-MB231 cells was observed by lfow cytometry. Conclusion：The overexpression of miR-340 can effectively inhibit the proliferation and increase the apoptosis of MDA-MB231 cells, which may be explained by up-regulating of protein cleaved-caspase-3 level.
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2014年第7期517-520,共4页
China Oncology
