何美玲,杨喆,戴研平,雷珊,高晓勤
HE Meiling,YANG Zhe,DAI Yanping,LEI Shan,GAO Xiaoqin

• 1:贵州医科大学基础医学院
• 2:贵州医科大学
• 3:遵义医药高等专科学校

摘要(Abstract)：

目的:探讨长链非编码RNA Linc00152对人胰腺癌细胞增殖能力的影响。方法:胰腺癌细胞株MIA PaCa-2和SW1990分为阴性对照组(NC组)和Linc00152干扰组(Si-Linc00152组),分别转染阴性对照(NC)及Si-Linc00152序列;细胞转染48 h后,采用实时荧光定量聚合酶链式反应(RT-qPCR)检测转染效率,CCK-8和平板克隆实验检测胰腺癌细胞的增殖能力,流式细胞术检测各组胰腺癌细胞的细胞周期,Western blot实验检测各组胰腺癌细胞cyclinD1和CDK4蛋白表达水平。结果:利用siRNA构建干扰模型48 h,Si-Linc00152组的Linc00152表达水平较NC组明显降低,差异有统计学意义(P<0.01); CCK-8实验结果显示,Si-Linc00152组细胞增殖较NC组显著下降,差异有统计学意义(P<0.01);平板克隆结果显示,Si-Linc00152组集落形成数目明显少于NC组,差异有统计学意义(P<0.01);流式细胞术结果显示干扰Linc00152后,细胞周期中的G1期百分比上升,差异有统计学意义(P<0.01);Western blot实验结果显示干扰Linc00152后,cyclinD1及CDK4蛋白表达水平均显著降低,差异有统计学意义(P<0.01)。结论:下调Linc00152可抑制胰腺癌细胞株MIA PaCa-2和SW1990的增殖,其机制可能与阻滞细胞周期、干扰cyclinD1及CDK4蛋白表达有关。
Objective: To investigate the effect of long-chain non-coding RNA Linc00152 on the proliferation of human pancreatic cancer cells. Methods: Pancreatic cancer cell lines MIA paca-2 and SW1990 were transfected with negative control(NC) and si-linc00152 sequence respectively. Transfection efficiency was measured by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR), the proliferation of pancreatic cancer cells was detected by CCK-8 and colony formation assay, the cell cycle of MIA PaCa-2 cells and SW1990 cells were detected by flow cytometry.Western blot assay was performed to detect the expression levels of CyclinD1 and CDK4 proteins in pancreatic cancer cells. Results: After knockdown of Linc00152, the expression of Linc00152 in Si-Linc00152 group was significantly lower than that in NC group(P<0.01). CCK-8 showed that the cell proliferation of Si-Linc00152 group was significantly lower than that of NC group(P<0.01). Colony formation assay showed that the number of colonies formed in the Si-Linc00152 group was significantly lower than that in the NC group(P<0.01). Flow cytometry results showed that the percentage of G1 phase in the cell cycle increased after interference with Linc00152(P<0.01). Western blot results showed that the protein expression levels of cyclinD1 and CDK4 were significantly decreased after interference with Linc00152(P<0.01). Conclusion: Down-regulation of Linc00152 inhibits the proliferation of pancreatic cancer cell lines MIA PaCa-2 and SW1990, which may be related to the inhibition of cell cycle and interference of cyclinD1 and CDK4 protein expressions.

关键词(KeyWords)： 胰腺肿瘤;长链非编码RNA;细胞增殖;转染;细胞周期蛋白
pancreatic neoplasms;carcinoma;long noncoding RNA;cell proliferation;transfection;cyclin

Abstract:

Keywords:

基金项目(Foundation): 贵州省科学技术基金[黔科合人才团队(2014)4005]

作者(Author): 何美玲,杨喆,戴研平,雷珊,高晓勤
HE Meiling,YANG Zhe,DAI Yanping,LEI Shan,GAO Xiaoqin

DOI: 10.19367/j.cnki.1000-2707.2019.12.010

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