MicroRNA⁃141靶向致癌基因Bmi⁃1抑制胰腺癌细胞的增殖
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江苏省卫计委面上项目(H201409);江苏省“六大人才高峰”资助(WAW?008)


miR⁃141;Bmi⁃1;细胞增殖;胰腺癌
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    摘要:

    目的:探讨miR-141调控胰腺癌细胞增殖的机制。方法:采用qRT-PCR检测正常胰腺细胞和4种胰腺癌细胞株中miR-141的表达。从Gene Expression Omnibus数据库(GEO)下载包含36例胰腺癌样本和16例正常样本的GSE71533 miR-141表达文件,分析miR-141在胰腺癌及癌旁的表达差异。Western blot 检测Bmi-1在正常胰腺细胞和4种胰腺癌细胞株中的表达。荧光素酶报告基因检测miR-141和Bm-1之间的靶向关系。通过CCK8、平板克隆形成检测Bmi-1和miR-141对胰腺癌细胞增殖能力的影响。结果:qRT-PCR显示与正常胰腺细胞相比,miR-141在胰腺癌细胞中表达下调;GSE71533数据集分析结果也显示miR-141在胰腺癌组织中下调;Western blot分析结果表明Bmi-1在胰腺癌细胞中高表达,双荧光素酶实验表明miR-141锚定在Bmi-1的3′非编码区,下调Bmi-1可以抑制胰腺癌细胞的增殖;上调miR-141可以降低Bmi-1的蛋白质表达水平,从而抑制胰腺癌细胞中细胞增殖。结论:miR-141通过靶向Bmi-1在胰腺癌中起抑制作用,在胰腺癌诊疗中具有潜在功能。

    Abstract:

    Objective:To investigate the mechanism of miR-141 regulating cell proliferation in pancreatic cancer. Methods:Expression of miR-141 in normal pancreas cell and four pancreatic cancer cell lines were detected by quantitative reverse transcription PCR(qRT-PCR). miR-141 expression file of GSE71533 which including 36 pancreatic cancer samples and 16 normal samples was downloaded from Gene Expression Omnibus(GEO)to analyze the expression of miR-141. Western blot was used to detect the expression of Bmi-1 in normal pancreatic cell and four pancreatic cancer cells Luciferase reporter assay was performed to analyze the relationship between Bmi-1 and miR-141. The effects of miR-141 and Bmi-1 on cell proliferation was examined by CCK8 and colony formation assay,respectively. Results:The qRT-PCR results showed that miR-141 was significantly down-regulated in pancreatic cancer cells. miR-141 was down-regulated in pancreatic cancer tissues by analyzing the microarray of GSE71533. Western blot analysis results demonstrated that Bmi-1 was up-regulated in pancreatic cancer cells. The dual luciferase assay indicated that miR-141 was anchored at the 3′-untranslated region of Bmi-1. Down-regulation of Bmi-1 inhibited the proliferation of pancreatic cancer cells. Upregulating miR-141 decreased the protein level of Bmi-1,thus repressing cell proliferation in pancreatic cancer cell. Conclusions:miR-141 acts as a tumor-suppressor in pancreatic cancer by targeting Bmi-1. These findings revealed that miR-141’s potential function in the treatment of pancreatic cancer.

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吴康健,吴假假,曾 凯,张晓辉,魏 威,许利剑. MicroRNA⁃141靶向致癌基因Bmi⁃1抑制胰腺癌细胞的增殖[J].南京医科大学学报(自然科学版),2018,(7):950-955

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  • 收稿日期:2018-03-08
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  • 在线发布日期: 2018-07-20
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