摘要
目的探讨miRNA-155对人乳腺癌MCF-7细胞TP53INP1表达及其增殖的影响。方法采用LipofectamineTM2000将miRNA-155 mimics(或带荧光无关序列阴性对照FAM-NC)转染雌激素受体(Estrogen receptor,ER)阳性人乳腺癌MCF-7细胞,并设空白对照组;RT-PCR法检测转染后miRNA-155的表达水平;CCK-8法检测miRNA-155对MCF-7细胞增殖能力的影响;流式细胞术检测MCF-7细胞的凋亡率和增殖周期;Western blot法检测TP53INP1、Caspase-3及p21蛋白的表达水平。结果与空白对照组和阴性对照组相比,转染组miRNA-155的表达量明显增高;随着时间的延长,MCF-7细胞增殖活力逐渐增加;G1期细胞明显减少,S和G2期细胞增多,凋亡率明显降低;TP53INP1、Caspase-3激活型及p21蛋白的表达水平均明显降低。结论 miRNA-155能够抑制人乳腺癌MCF-7细胞TP53INP1蛋白的表达,进而抑制肿瘤细胞凋亡、促进细胞增殖,在乳腺癌发生发展中具有重要作用。
Objective To investigate the effects of miRNA-155 on expression of TP53INPI protein and proliferation of human breast cancer MCF-7 cells. Methods Estrogen receptor （ER） positive MCF-7 cells were transfected with miRNA-155 mimics in mediation of LipofectamineTM2000, using those untransfected as blank control and those transfected with FAM- NC as negative control, and determined for expression level of miRNA-155 by RT-PCR, while the proliferation level by CCK8 method, the apoptosis rate and cell cycle of by flow cytometry, and the expression levels of TP53INP1, Caspase-3 and p21 protein by Western blot. Results The expression level of miR-155 in MCF-7 cells transfected with miRNA-155 mimics was significantly higher than those in blank and negative control groups. The proliferation activity of MCF-7 cells increased with the increasing hours after transfection, while the cells at G1 phase decreased, and those at S and G2 phases increased, and the apoptosis rate of cells decreased significantly. The expression levels of TP53INP1, Caspase-3 and P21 decreased significantly. Conclusion The miRNA-155 inhibited the expression of TP53INP1 in MCF-7 cells so as to inhibit the apoptosis and promote the proliferation of tumor cells, which played an important role in the onset and progress of breast cancer.
出处
《中国生物制品学杂志》
CAS
CSCD
2013年第9期1273-1277,共5页
Chinese Journal of Biologicals
作者简介
通讯作者：邓华瑜，E-mail：cqdenghy@yahoo．com．cn
