miR-3619-5p对乳腺癌细胞MCF-7和T47D增殖、凋亡的影响及分子机制

发布时间：2018-06-22 10:38

本文选题：乳腺肿瘤 + miR--p　；参考：《临床与实验病理学杂志》2017年11期

【摘要】：目的探讨miR-3619-5p转染对乳腺癌细胞系MCF-7和T47D增殖和凋亡的影响及可能的分子机制。方法乳腺癌培养至对数生长期,根据对其不同处理分为2组:对照组(转染ds Control)和实验组(转染miR-3619-5p)。5-乙炔基-2'脱氧尿嘧啶核苷(Ed U)增殖实验和集落形成实验检测乳腺癌细胞增殖能力变化;流式细胞术(flow cytometry,FCM)检测乳腺癌细胞周期分布和凋亡情况;实时定量聚合酶链反应(qRT-PCR)检测p21、CDK6和Cyclin D1 mRNA的表达;Western blot法检测p21、CDK6和Cyclin D1蛋白的表达变化。结果与dsControl组相比,转染miR-3619-5p的乳腺癌细胞增殖能力显著降低(P0.05)。转染miR-3619-5p后,位于G_0/G_1期的细胞比例明显升高,位于S期和G_2/M期的细胞比例明显降低,细胞凋亡率显著上升。与ds Control组相比,转染miR-3619-5p后两种细胞系中p21 mRNA均明显上调(P0.01),而CDK6和Cyclin D1 mRNA的表达均明显下调(P0.01)。Western blot检测结果与qRT-PCR结果一致。结论 miR-3619-5p可显著抑制乳腺癌细胞的增殖能力,促进细胞凋亡,其分子机制可能是通过激活乳腺癌细胞中抑癌基因p21的表达。
[Abstract]:Objective to investigate the effect of miR-3619-5p transfection on the proliferation and apoptosis of breast cancer cell line MCF-7 and T47D and its possible molecular mechanism. Methods Breast cancer was cultured to logarithmic growth stage. According to the different treatments, the breast cancer cells were divided into two groups: control group (transfected DS Control) and experimental group (transfected miR-3619-5p) .5-acetyl-2'deoxyuridine (Ed U) proliferation test and colony forming assay to detect the proliferation ability of breast cancer cells. Flow cytometry (flow) was used to detect the cell cycle distribution and apoptosis, and the expression of p21 CDK6 and Cyclin D1 mRNA was detected by real-time quantitative polymerase chain reaction (qRT-PCR). Western blot was used to detect the expression of p21 CDK6 and Cyclin D1 protein. Results compared with dsControl group, the proliferation ability of breast cancer cells transfected with miR-3619-5p was significantly decreased (P0.05). After transfection of miR-3619-5p, the percentage of cells in G0 / GHS 1 phase was significantly increased, while that in S phase and G2 / M phase was significantly decreased, and the apoptosis rate was significantly increased. Compared with DS Control group, p21 mRNA was up-regulated (P0.01) in the two cell lines transfected with miR-3619-5p, while the expression of CDK6 and Cyclin D1 mRNA was significantly down-regulated (P0.01). The results of Western blot were consistent with those of qRT-PCR. Conclusion miR-3619-5p can significantly inhibit the proliferation of breast cancer cells and promote cell apoptosis. The molecular mechanism of miR-3619-5p may be by activating the expression of tumor suppressor gene p21 in breast cancer cells.
【作者单位】：河南大学附属郑州市妇幼保健院病理科;
【分类号】：R737.9

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