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目的研究microRNA-133b在结直肠癌细胞中的表达及其对细胞迁移和侵袭能力的影响。方法采用实时定量PCR检测microRNA-133b(miR-133b)在不同恶性程度的结直肠癌细胞及正常结肠细胞中的表达差异;构建miR-133b的真核表达载体并稳定转染SW-620细胞,筛选出稳定表达miR-133b及对照质粒的细胞株SW-620-133b和SW-620-HK,实时定量PCR检测转染后miR-133b表达的变化;划痕实验检测miR-133b对SW-620细胞迁移能力的影响;Transwell侵袭实验检测miR-133b对SW-620细胞侵袭能力的影响。结果正常结肠细胞系CCD-18Co中miR-133b的表达显著高于4种结直肠癌细胞(SW-480,HT-29,LoVo,SW-620)(P<0.05),且其在结直肠癌细胞中的表达随着肿瘤细胞恶性程度的增加而降低。稳定转染后,SW-620-133b细胞miR-133b表达量较SW-620及SW-620-HK组显著增高(P<0.05);miR-133b对SW-620细胞迁移和侵袭能力有显著的抑制作用,抑制率分别为(42.2±2.6)%及(66.3±8.7)%。结论miR-133b在结直肠癌细胞中呈低表达,miR-133b高表达能显著抑制SW-620细胞的迁移和侵袭能力。
Objective To study the expression of microRNA-133b in colorectal cancer cells and its effect on cell migration and invasion. Methods Real-time quantitative PCR was used to detect the expression of microRNA-133b (miR-133b) in colorectal cancer cells and normal colorectal cancer cells with different degrees of malignancy. The eukaryotic expression vector of miR-133b was constructed and stably transfected into SW-620 cells. The cell lines SW-620-133b and SW-620-HK stably expressing miR-133b and the control plasmid were screened, and the changes of miR-133b expression after transfection were detected by real-time quantitative PCR. The effect of miR-133b on the invasive ability of SW-620 cells was detected by Transwell invasion assay. Results The expression of miR-133b in normal colorectal cell line CCD-18Co was significantly higher than that in four colorectal cancer cells (SW-480, HT-29, LoVo and SW-620) The expression of the cells decreased with the malignant degree of tumor cells. After stable transfection, the expression of miR-133b in SW-620-133b cells was significantly higher than that in SW-620 and SW-620-HK groups (P <0.05); the ability of miR-133b on migration and invasion of SW-620 cells was significantly The inhibitory rates were (42.2 ± 2.6)% and (66.3 ± 8.7)%, respectively. Conclusion miR-133b is overexpressed in colorectal cancer cells. High expression of miR-133b significantly inhibits the migration and invasion of SW-620 cells.