FOXJ1调控AKT信号通路对结直肠癌细胞凋亡和侵袭能力的影响

目的 探讨叉头框转录因子J1(FOXJ1)对人结直肠癌细胞凋亡和侵袭能力的影响及机制.方法 应用Western Blot方法检测人结直肠癌细胞HCT116、HT29、SW480及人正常肠上皮细胞FHC中FOXJ1的表达水平.细胞中转染pEGFP-N1-FOXJ1和pEGFP-N1,Western Blot检测细胞中FOXJ1、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、基质金属蛋白酶-2(MMP-2)、蛋白激酶B(AKT)、磷酸化的蛋白激酶B(p-AKT)的表达水平,流式细胞仪检测细胞的凋亡情况,Transwell小室检测细胞的侵袭能力.结果 人结直肠癌细胞HCT116、HT29、SW480中FOXJ1的表达水平均明显低于人正常肠上皮细胞FHC(P﹤0.01).人结直肠癌细胞HCT116中FOXJ1的表达水平下降最多,后续选用HCT116细胞继续研究.转染pEGFP-N1后的细胞凋亡率、侵袭细胞数目及细胞中FOXJ1、Cleaved Caspase-3、MMP-2、AKT、p-AKT的表达水平与对照组比较,差异均无统计学意义(P﹥0.05).转染pEGFP-N1-FOXJ1后的细胞凋亡率及细胞中FOXJ1、Cleaved Caspase-3的表达水平明显高于对照组(P﹤0.01);转染pEGFP-N1-FOXJ1后的侵袭细胞数目及细胞中MMP-2、p-AKT的表达水平明显低于对照组(P﹤0.01).结论 FOXJ1在人结直肠癌细胞中过表达.FOXJ1能够促进结直肠癌细胞凋亡,抑制结直肠癌细胞侵袭,其作用机制可能与AKT信号通路有关.

Effects of FOXJ1 regulated AKT signaling pathway on apoptosis and invasion of colorectal cancer cells

Objective To investigate the effect and mechanism of FOXJ1 on apoptosis and invasion of colorectal can-cer cells. Method The expression level of FOXJ1 was detected by Western Blot in human colorectal cancer cells HCT116, HT29, SW480 and normal intestinal epithelial cells FHC. pEGFP-N1-FOXJ1 and pEGFP-N1 were transfected into cells, detected the levels of FOXJ1, Cleaved Caspase-3, MMP-2, AKT, p-AKT by Western Blot,detected cell apopto-sis by flow cytometry, detected cell invasion ability by Transwell. Result The expression of FOXJ1 in human colorectal cancer cells HCT116, HT29, SW480 was significantly lower than that of human normal intestinal epithelial cells FHC (P<0.01). The expression level of FOXJ1 in colorectal cancer cell line HCT116 was decreased more, and the HCT116 for the continued study. Cell transfected with pEGFP-N1 cell apoptosis rate, the number of invasive cells and FOXJ1, Cleaved Caspase-3, MMP-2, AKT, p-AKT expression levels had no significantly difference compared with the control group (P>0.05). Cell transfected with pEGFP-N1-FOXJ1 apoptosis rate and FOXJ1, Cleaved Caspase-3 expression levels were sig-nificantly higher than the control group (P<0.01). Cell transfected with pEGFP-N1-FOXJ1 invasion cells number and MMP-2, p-AKT expression levels were significantly lower than the control group (P<0.01). Conclusions Overexpres-sion of FOXJ1 in human colorectal cancer cells. FOXJ1 can promote the apoptosis of colorectal cancer cells and inhibit the invasiveness of colorectal cancer cells. The mechanism may be related to AKT signaling pathway.