霍小蕾, 裴振, 李永芝, 等.长链非编码RNAH19促进结直肠癌细胞株增殖的研究.四川大学学报(医学版),2017,48(6):844-849
长链非编码RNAH19促进结直肠癌细胞株增殖的研究
Effects of LncRNA H19 on Proliferation of Human Colorectal Cancer SW620 Cells
  
中文关键词:  lncRNA结直肠癌细胞增殖治疗
英文关键词:Long non-coding RNAColorectal cancerCell proliferationTherapy
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中文摘要:
      目的 研究长链非编码RNA(long non-coding RNA, lncRNA) 家族分子母源性印迹基因19 转录因子(H19 )在结直肠癌组织及细胞株中的表达情况及其对人结直肠癌SW620细胞增殖的影响。方法 采用实时荧光定量PCR(qRT-PCR)检测20例结直肠癌组织及其癌旁组织,以及结直肠癌细胞株SW480、HCT116、SW620和正常结直肠上皮NCM460细胞中H19的表达。结直肠癌SW620细胞分别转染H19 siRNA(si-H19 组) 和阴性对照序列(si-NC组) ,分别采用流式细胞技术、细胞克隆形成实验和CCK8检测两组细胞的增殖情况。qRT-PCR和Western blot检测两组细胞G1/S-特异性周期蛋白-D1(CyclinD1)和细胞周期素依赖激酶(cyclin dependent kinase 4, CDK4)的表达情况。结果 与癌旁组织和正常结直肠上皮NCM460细胞相比较,结直肠癌组织和细胞株中H19的表达水平升高( P<0.05);si-H19组细胞H19表达水平低于si-NC组,且细胞活力和细胞克隆数均低于si-NC组,以上差异均有统计学意义( P<0.05);同时,相较于si-NC组,si-H19组CyclinD1和CDK4基因和蛋白的表达受到了明显抑制( P<0.05)。结论 结直肠癌组织及细胞株中H19 呈高表达,沉默H19 表达能明显抑制结直肠癌细胞的增殖能力,为结直肠癌的靶向治疗提供了潜在策略。
英文摘要:
      ObjectiveTo determine the expression level of long non-coding RNA (lncRNA) imprinted maternallyexpressed transcript (H19) in colorectal cancer tissues and its effect on proliferation of colorectal cancer SW620 cells. MethodsReal-time quantitative PCR (qRT-PCR) was applied to detect the expression of H19 in 20 paired tumor tissues and adjacent normal tissues, and in normal NCM460 cells and colorectal cancer SW480, HCT116 and SW620 cells. The specific small interfering RNA for H19 (si-H19 group) or negative control sequence (si-NC group) were transfected into SW620 cells. Proliferation of the transfected cells was detected using flow cytometry, CCK8 assay and clone formation experiment. The expressions of CyclinD1 and cyclin dependent kinase 4 (CDK4) were detected by qRT-PCR and Western blot. ResultsThe expression levels of H19 in colorectal cancer tissues and cells were higher compared with those in adjacent normal tissues and normal NCM460 cells. Lower H19 level, cell activities and cell clone numbers were found in si-H19 transfected cells compared with those in si-NC transfected cells ( P<0.05). si-H19transfected cells had decreased expression of CyclinD1 and CDK4 ( P<0.05). ConclusionH19expression in colorectal cancer is high. Knock-down H19 expression can inhibit proliferation of colorectal cancer cells, which provides a potential strategy for targeted therapy of colorectal cancer.
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