摘要
目的探讨赖氨酸特异性组蛋白去甲基化酶1(KDM1A)对结直肠癌细胞增殖侵袭及AKT信号通路的影响。方法采用LipofectamineTM2000转染试剂将si-KDM1A转染至结直肠癌HCT15细胞中,MTT法检测细胞增殖能力,流式细胞仪检测细胞的周期变化,Transwell小室实验检测细胞的侵袭能力,RT-PCR检测细胞中KDM1A mRNA的表达,Western blotting检测KDM1A蛋白和AKT信号通路相关蛋白p21、Cyclin D1、MMP-2和p-AKT的表达。结果转染si-KDM1A后,HCT15细胞中KDM1A mRNA、KDM1A、Cyclin D1、MMP-2和p-AKT蛋白表达均显著降低,而p21蛋白表达显著升高;细胞的增殖能力和侵袭能力明显减弱;细胞在G1期所占比例明显增加,而在S期和G2期明显减少。结论下调KDM1A基因可抑制结直肠癌HCT15细胞的增殖、侵袭能力,其分子机制可能与抑制AKT信号通路的活化有关。
Objective To investigate the effects of lysine specific histone demethylation enzyme 1(KDM1A)on proliferation,invasion and AKT signaling pathway in colorectal cancer cells.Methods si-KDM1A was transfected into HCT15cells by LipofectamineTM 2000transfection reagent,and the proliferation ability was detected by MTT.Cell cycle changes were detected by flow cytometry,and cell invasiveness was detected by Transwell chamber assay.The expression of KDM1A mRNA in the cells was measured by RT-PCR,and the expressions of KDM1A protein and AKT signaling pathway related proteins p21,Cyclin D1,MMP-2and p-AKT were examed by Western blotting.Results After transfection of si-KDM1A,the expressions of KDM1A mRNA and KDM1A,Cyclin D1,MMP-2and p-AKT protein in HCT15cells were all decreased significantly,but the expression of p21protein was increased markedly ;the proliferation and invasion abilities of the cells were inhibited obviously ;the proportion of cells in G1 stage was increased,but decreased obviously in S stage and G2stage.Conclusion Down regulation of KDM1A gene can inhibit the proliferation and invasion of colorectal cancer HCT15cells,and its molecular mechanism may be related to inhibiting the activation of AKT signaling pathway.
作者
潘春鹏
李峰
PAN Chunpeng;LI Feng(Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200433,China)
出处
《胃肠病学和肝病学杂志》
CAS
2018年第12期1372-1376,共5页
Chinese Journal of Gastroenterology and Hepatology
