微小RNA-196a在结肠癌组织中表达及小干扰RNA沉默对结肠癌细胞侵袭和转移的影响

Expression of microRNA - 196a in colon cancer tissues and effects of small interfering RNA silen- cing on invasion and metastasis of colon cancer cells

目的观察微小RNA（miRNA，miR）-196a在结肠癌组织中表达及小f扰RNA（siRNA）沉默对结肠癌细胞侵袭和转移的影响。方法选取结肠癌患者72例，培养人结肠癌细胞株SW480，分为siRNA组、阴性对照组和空白对照组，实时荧光定量聚合酶链反应（FQ—PCR）检测结肠癌及癌旁组织和不同转染组SW480细胞中miR-196a表达，噻唑蓝（MTF）比色法检测细胞增殖结果，Transwell细胞迁移实验和侵袭实验检测细胞迁移和侵袭能力。结果结肠癌组织中miR-196a表达量为2．15±0．37，显著高于癌旁组织中的1．24±0．23，差异有统计学意义（P〈0．001）；结肠癌组织中miR-196a表达与TNM分期、淋巴结转移、远处转移和Dukes分期有关（P〈0．05）；siRNA组细胞中miR-196a表达量（0．67±0．12）显著低于阴性对照组和空白对照组，分别为0．95±0．17和0．934-0．14，差异有统计学意义（P〈0．05）；siRNA组细胞转染24、48、72、96h时吸光度（A）值均低于阴性对照组和空白对照组，差异均有统计学意义（P〈0．05）；Transwell小室检测结果显示，siRNA组细胞迁移细胞数和侵袭细胞数均低于阴性对照组和空白对照组，差异均有统计学意义（P〈0．05）。结论miR-196a在结肠癌组织中呈高表达，特异性沉默结肠癌细胞中miR-196a可抑制细胞增殖、侵袭和转移。

Objective To investigate the expression of microRNA （miRNA, miR） - 196a in co- lon cancer tissues and effects of siRNA silencing on invasion and metastasis of colon cancer cells. Methods Seventy - two cases of colon cancer patients were selected. Human colon cancer cell line SW480 cells were cultured and divided into small interfering RNA （siRNA） group, negative control group and blank control group. The expression levels of miR - 196a in colon cancer and adjacent tissues and SW480 cells in differ- ent transfected groups were detected by real -time fluorescent quantitative polymerase chain reaction （FQ -PCR）. The cell proliferations were tested by methyl thiazol tetrazolium （MTF） assay. The cell mi- grations and invasions were examined by Transwell migration assay and cell invasion assay respectively. Re- suits The expression levels of miR - 196a in colon cancer tissues were 2. 15 ± 0. 37, which were signifi- cantly higher than 1.24 ±0.23 in adjacent tissues （P 〈0. 01 ）. The expression levels of miR - 196a in co- lon cancer tissues were related with TNM stage, lymph node metastasis, distant metastasis and Dukes stage （P 〈0. 05）. The expression levels of miR- 196a in siRNA group were 0. 67 ±0. 12, which were signifi- cantly lower than 0. 95 ± 0. 17 in the negative control group and 0. 93 ± 0. 14 in the blank control groups （ P 〈 0.05 ）. The absorbance （A） values at 24, 48, 72 and 96 h in siRNA group were significantly lower than in negative control group and blank control group （P 〈 0.05 ）. Transwell chamber assay results showed the number of migration and invasion cells in siRNA group was significantly less than in negative control group and blank control group （ P 〈 0.05 ）. Conclusion MiR - 196a was highly expressed in colon cancer tissues. Specific silencing of miR - 196a in colon cancer cells could inhibit cell proliferation, invasion and metastasis.