小干扰RNA沉默赖氨酸去甲基化酶2A对结肠癌细胞增殖及侵袭能力的影响

The effect of silencing lysine （K） - specific demethylase 2A by small interference RNA on prolifera- tion and invasion capability of colonic cancer cell line

目的观察小干扰RNA（siRNA）沉默赖氨酸去甲基化酶2A（KDM2A）基因对结肠癌细胞株LoVo增殖及侵袭能力的影响。方法构建针对KDM2A的特异性siRNA（siKDM2A），瞬时转染LoVo细胞，利用实时定量聚合酶链反应（Real—timePCR）技术和Westernblot技术检测LoVo细胞中KDM2A基因表达；细胞计数试剂盒（CCK_8）增殖实验检测LoVo细胞转染siKDM2A后细胞增殖能力的变化；侵袭实验观察低表达KDM2A对LoVo细胞侵袭能力的影响；Westernblot法检测LoVo细胞低表达KDM2A后增殖细胞核抗原（PCNA）表达。结果Real—timePCR和Westernblot法验证siKDM2A转染成功；CCK一8增殖实验中，KDM2A低表达可抑制LoVo细胞的增殖能力；侵袭实验中低表达KDM2A细胞穿膜数目（33．0±3．3）个，较Blank组[（91．0±3．3）个]和siControl[（87．7±2．9）个]明显减少（P〈0．05）；低表达KDM2A细胞中PCNA表达量较对照组明显降低。结论低表达KDM2A基因可抑制结肠癌细胞株LoVo的增殖能力和侵袭能力。

Objective To investigate the effects of small interference RNA （siRNA） targeting 1y- sine （K） - specific demethylase 2A （KDM2A） gene on proliteration and invasion of colonic cancer cell line LoVo. Methods Synthesized siRNA targeting KDM2A （siKDM2A） was transfected into LoVo ceils, then the expression of KDM2A mRNA and protein were detected by real - time quantitative polymerase chain reaction （ Real - time PCR） and Western blotting. The cell viability was detected by cell counting kit - 8 （ CCK - 8） and the cell invasion was examined by Transwell assay. Results Compared to the con- trol, the expression of KDM2A was significantly decreased in the experimental group and the expression of proliferating cell nuclear antigen （PCNA） was significantly decreased at the same time. In the proliferation assay, the proliferation ability of the experimental group was significantly decreased than the control group. In the migration assay, the number of penetrating the membrane was markedly decreased in the experimen- tal group as compared with the control. Conclusion Down - regulation of KDM2A could inhibit the prolif- eration and invasion capability of human colonic cancer cell LoVo.