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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 effect of miRNA-155 on the expression of TP53INP1 and its proliferation in human breast cancer MCF-7 cells. Methods Human breast cancer MCF-7 cells were transfected with miRNA-155 mimics (or FAM-NC with fluorescent unrelated sequence) by LipofectamineTM2000, and blank control group was established. RT-PCR The effect of miRNA-155 on the proliferation of MCF-7 cells was detected by CCK-8 assay; the apoptosis rate and proliferation cycle of MCF-7 cells were detected by flow cytometry; the expression of miRNA- The levels of TP53INP1, Caspase-3 and p21 protein were detected. Results Compared with the blank control group and the negative control group, the expression of miRNA-155 in the transfected group was significantly increased. With the prolongation of time, the proliferation activity of MCF-7 cells gradually increased; the cells in G1 phase decreased significantly; the cells in S and G2 phase Increased, the apoptosis rate was significantly reduced; TP53INP1, Caspase-3 activation type and p21 protein expression levels were significantly lower. Conclusion miRNA-155 can inhibit the expression of TP53INP1 protein in human breast cancer MCF-7 cells, and then inhibit the apoptosis of tumor cells and promote cell proliferation, which play an important role in the development of breast cancer.