出国医疗

miR-301在结肠癌细胞增殖和凋亡中的作用

来源:万方数据    时间:2014-02-10 15:13  浏览次数:

  The role of miR-301 in colon cancer cell proliferation and apoptosis

        【作者】
张忠来、熊欢、张兵、朱培谦

        【关键词】miR-301、结肠癌、反义单核苷酸

  背景与目的:各种恶性肿瘤中的miR-224都存在过度表达的问题,然而它在结肠癌中的表达及功能还不清楚。本研究旨在探究miR-301在结肠癌组织中的基因表达,并在体内外研究miR-301 (ASO)对结肠癌细胞增殖和凋亡的控制效果。方法:运用实时定量PCR 分析120例结肠癌患者组织及对应的癌旁组织中miR-301的表达;miR-301ASO转染后,使用MTT化验、菌落形成实验、流式细胞技术及体内实验来测量miR-301在SW620细胞中的生物效应。

  结果:在120例结肠癌病例中,63.33%(76/120) miR-301存在过度表达。与对照组miR-301的表达量 (0.50±0.07,P=0.00)相比,miR-301在SW620细胞中(与miR-301 ASO发生转染,0.09±0.01)的表达显著偏低;MTT化验结果显示,与miR-301 ASO(P=0.00)转染后,SW620细胞的24、48、96 小时存活数量均明显低于对照组;菌落形成实验结果显示,miR-301 ASO组菌落形成率(5.33%±0.74%) 较对照组(33.33%±8.38%,P=0.00)显著偏低。体内研究进一步证实miR-301 ASO可以抑制SW620细胞的增殖,而miR-301 ASO组生长较对照组明显偏慢(P=0.00)。流式细胞仪检测显示,转染miR-301 ASO组SW620细胞凋亡指数 (15.68±1.46)较对照组(3.36±0.88,P=0.02)明显偏高;另外,降低miR-301的表达后,发现Bcl2 mRNA和Bcl2蛋白均明显下降(P=0.00,P=0.00)。

  结论:miR-301在结肠癌组织中是过度表达的,降低miR-301的表达可有效抑制结肠癌细胞生长、促进细胞凋亡。miR-301可能成为结肠癌基因表达调控的新靶点。

  Abstract:

  Background and purpose: The miR-224 in a variety of malignant tumors is overexpression, however, its expression and function in colon cancer are not clear. The aim of this study was to investigate the expression of miR-301 in colon cancer tissues and demonstrate the regulative effects of miR-301 ASO on the proliferation and apoptosis of colon cancer cell in vitro and in vitro. Methods:The expression of miR-301 in 120 colon cancer tissues and their adjacent tissues was detected by real-time quantitative PCR method. After transfection with miR-301ASO, the biological effects of miR-301 in SW620 cells were measured by MTT assay, the colony formation experiment, flow cytometry and the in vivo experiment.

  Results: The expression level of miR-301 was found to be over expressed in 63.33% (76/120) of the colon cancer cases (P<0.05). miR-301 expression in SW620 cells (transfection with miR-301 ASO, 0.09±0.01) was significantly less than control group (0.50±0.07, P=0.00). MTT assay results showed that SW620 cells survived rate at 24, 48 and 96 h decreased greatly after transfection with miR-301ASO (P=0.00). Clone formation assay revealed that miR-301 ASO group colony formation rate (5.33%±0.74%) was significantly lower than the control group (33.33%±8.38%, P=0.00). In vivo study further confirmed that miR-301ASO could inhibit the proliferation of SW620 cells (P<0.05), and miR-301ASO group grew substantially slow compared with the negative control group (P=0.00). Flow cytometry indicated that the apoptotic index in miR-301 ASO group (15.68±1.46) was significantly higher than the control group (3.36±0.88, P=0.02). In addition, the Bcl2 mRNA and protein were significantly decreased after reduce the expression of miR-301 (P=0.00, P=0.00). Conclusion:MiR-301 was overexpressed in human colon cancer. Reduce the expression of miR-301 can effectively inhibit the growth of colon cancer cells and promote apoptosis. MiR-301 may become a new target for the regulation of gene expression in colon

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