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目的:探讨微小RNA-338-3p(miR-338-3p)对人结直肠癌细胞侵袭迁移能力及Smoothened(SMO)蛋白表达的影响。方法:应用实时荧光定量PCR(Real-time PCR)检测四种人结肠癌细胞系Lovo、HT-29、HCT116、SW620中miR-338-3p表达状况,并以人脐静脉内皮细胞(HUVEC)作为对照。采取脂质体转染miR-338-3p前体(pre-miR-338-3p)至人结直肠癌细胞系SW620,观察细胞转染前后miR-338-3p的表达变化,Transwell侵袭实验及划痕实验检测转染前后细胞侵袭及迁移能力的改变;以Western-blotting和RT-PCR分析SMO蛋白及mRNA表达状况。结果:检测四种结直肠癌细胞系中miR-338-3p表达水平均较正常对照细胞显著降低,差异具有统计学意义(P<0.01);SW620细胞转染pre-miR-338-3p后,miR-338-3p表达水平升高,SMO蛋白表达下降,同时肿瘤细胞侵袭、迁移能力明显增强,与对照组相比差异具有统计学意义(P<0.01);但SMO mRNA表达水平在转染前后差异无统计学意义(P>0.05)。结论:MiR-338-3p可通过抑制结直肠癌细胞系中SMO蛋白表达从而抑制肿瘤细胞侵袭转移。
Objective: To investigate the effect of microRNA-338-3p (miR-338-3p) on the invasion and migration of human colorectal cancer cells and the expression of Smoothened (SMO) protein. Methods: Real-time PCR was used to detect the expression of miR-338-3p in four human colon cancer cell lines, Lovo, HT-29, HCT116 and SW620. Human umbilical vein endothelial cells (HUVECs) Control. The miR-338-3p precursor (pre-miR-338-3p) was transfected into human colorectal cancer cell line SW620 by lipofectamine 2000. The expression of miR-338-3p before and after transfection was observed. Scratch assay was used to detect the changes of cell invasion and migration before and after transfection. The expression of SMO protein and mRNA was analyzed by Western-blotting and RT-PCR. Results: The expression levels of miR-338-3p in four colorectal cancer cell lines were significantly lower than those in normal control cells (P <0.01). After pre-miR-338-3p transfection, The expression of miR-338-3p increased and the expression of SMO protein decreased. Meanwhile, the invasion and migration of tumor cells were significantly enhanced compared with the control group (P <0.01) The difference was not statistically significant (P> 0.05). Conclusion: MiR-338-3p can inhibit tumor cell invasion and metastasis by inhibiting SMO protein expression in colorectal cancer cell lines.