临床肿瘤学杂志

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miR-203在结直肠癌中的表达及其功能分析

李丹1,周连帮2,王年飞3,张道权4   

  1. 1 安徽医科大学第二附属医院科教部 2 安徽医科大学第二附属医院普外科 3 安徽医科大学第二附属医院肿瘤科 4 安徽医科大学第二附属医院消化内科
  • 收稿日期:2015-03-11 修回日期:2015-04-06 出版日期:2015-05-31 发布日期:2015-05-31

The expression and function of microRNA-203 in colorectal cancer

LI Dan, ZHOU Lianbang, WANG Nianfei, ZHANG Daoquan.   

  1. the Second Affiliated Hospital of Anhui Medical University
  • Received:2015-03-11 Revised:2015-04-06 Online:2015-05-31 Published:2015-05-31

摘要: 目的 探讨microRNA-203(miR203)在结直肠癌中的表达情况及其临床意义,并在结直肠癌细胞中验证其功能。方法 采用实时荧光定量PCR(qRT-PCR)检测52例结直肠癌及对应癌旁组织中miR-203的水平,分析miR-203表达与临床病理参数及术前癌胚抗原的关系;检测miR203在3种结直肠癌细胞株SW480、Lovo和HT29及人正常结肠黏膜上皮细胞NCM460中的表达情况,选取miR-203水平最低的细胞分别转染miR-203模拟物(mimics)和miR-203 control,采用MTT法、流式细胞仪PI/Annexin V双染法和Transwell法检测转染miR203对细胞增殖、凋亡和侵袭能力的影响。结果 结直肠癌组织中miR-203的相对表达量为0.372±0.019,低于癌旁组织的miR-203水平,且其表达与TNM分期、淋巴结转移、肿瘤大小及术前癌胚抗原水平均有关(P<0.05);与NCM460细胞相比(其miR-203表达水平设为1.00),结直肠癌细胞SW480、Lovo和HT-29的miR203相对表达量依次为0.26±0.07、0.44±0.05和0.32±0.04,选取SW480细胞进行后续实验。MTT检测显示,转染miR-203 mimics 48、72和96 h的SW480细胞吸光值均低于转染miR-203 control组,差异均有统计学意义(P<0.05);且转染miR-203 mimics 48和96 h的SW480细胞凋亡率高于转染miR-203 control组,但穿膜细胞数低于转染miR-203 control组(P<0.05)。结论 miR-203在结直肠癌组织和细胞中低表达,上调miR203水平可抑制结直肠癌细胞的增殖、侵袭并诱导凋亡。

Abstract: Objective To explore the expression and clinical significance of microRNA203 (miR-203) in colorectal cancer, and to verify its function in colorectal carcinoma cells. Methods The realtime quantitative PCR (qRT-PCR) was used to detect the levels of miR-203 in 52 cases of colorectal cancer and corresponding adjacent tissues. The relationship between the levels of miR203 and clinical pathological parameters (gender, age, tumor size, tumor location, tumor differentiation, clinical stage and lymph node metastasis) and preoperative carcinoembryonic antigen was carried out. The levels of miR203 in three common colorectal cancer cell lines (SW480, Lovo and HT-29) and human normal colonic mucosal epithelial cells NCM460 were also measured via qRTPCR. The cell with the lowest miR-203 level was chosen and then subjected to the transfection with miR-203 analogue (mimics) and miR-203 control, respectively. The influences of miR-203 mimics on the cell proliferation, invasion and apoptosis were evaluated by MTT method, flow cytometry with PI/Annexin V double staining and Transwell, respectively. Results The relative expression of miR-203 in colorectal cancer tissues was 0.372±0.019, lower than that in paracarcinoma tissues. The levels of miR-203 were related with TNM staging, lymph node metastasis, tumor size and preoperative carcinoembryonic antigen levels. When the miR203 level in NCM460 cell was taken as reference (1.00), the relative expression of miR-203 in SW480, Lovo and HT-29 cells were 0.26±0.07, 0.44± 0.05 and 0.32±0.04, respectively. As shown in the MTT analysis, the absorption values of miR-203 mimics group were lower than the miR203 control group at 48, 72 and 96 h (P<0.05). Moreover, there were higher apoptotic rates but lower transmembrane cell numbers in miR-203 mimics group versus miR203 control group at 48 and 96 h. Conclusion There are low levels of miR-203 in both colorectal cancer tissues and cells. The elevation of miR-203 can inhibit the cell proliferation and invasion but induce the apoptosis in colorectal cancer.

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