慢病毒介导的 FLOT 2基因沉默影响结肠癌细胞生物学功能的影响及机制  

Lentivirus-mediated gene silencing of FLOT2 affects the proliferation,apoptosis,migration and invasion of colon cancer cells

作　　者：伍班名 张涛[2] 张秀容[1] 钟寒迪[3] 王桂林[1] WU Banming;ZHANG Tao;ZHANG Xiurong;ZHONG Handi;WANG Guilin(Emergency Department,The Second People′s Hospital of Yibin,Yibin 644000,Sichuan,China;Laboratory Department,The Second People′s Hospital of Yibin,Yibin 644000,Sichuan,China;Gastrointestinal Surgery,The Second People′s Hospital of Yibin,Yibin 644000,Sichuan,China)

机构地区：[1]宜宾市第二人民医院急诊科,四川宜宾644000 [2]宜宾市第二人民医院检验科,四川宜宾644000 [3]宜宾市第二人民医院胃肠外科,四川宜宾644000

出　　处：《西部医学》2022年第4期530-535,共6页Medical Journal of West China

摘　　要：目的构建干扰FLOT2表达的慢病毒载体,探讨抑制FLOT2表达对结肠癌细胞增殖、凋亡、侵袭、迁移的影响及机制。方法通过qRT-PCR检测结直肠癌组织及人结肠癌LoVo、SW480及HT29细胞FLOT2的表达水平,Western blot检测结肠癌细胞FLOT2表达。将含FLOT2 siRNA的慢病毒载体转染结肠癌LoVo细胞和SW480细胞。采用qRT-PCR及Western blot检测转染后的细胞FLOT2表达。MTT法、流式细胞术、Transwell小室、Western blot分别检测细胞增殖能力、凋亡率、侵袭迁移能力及PCNA、cleaved caspase3、E-cadherin和Vimentin蛋白水平。结果结直肠癌组织FLOT2 mRNA表达水平明显高于癌旁组织,三个结肠癌细胞FLOT2 mRNA及蛋白水平均明显高于在正常结肠上皮细胞NCM460(P<0.05)。LoVo细胞和SW480细胞转染FLOT2 siRNA慢病毒载体后,FLOT2 mRNA及蛋白表达水平均明显降低,且可明显降低细胞增殖、侵袭和迁移能力,促进细胞凋亡,下调PCNA和Vimentin表达,上调cleaved caspase3和E-cadherin(P<0.05)。结论抑制FLOT2表达可明显降低结肠癌细胞增殖、侵袭和迁移能力,促进细胞凋亡,其机制可能与调节PCNA、cleaved caspase3、E-cadherin和Vimentin表达有关。Objective To construct lentivirus vector that interferes with FLOT2 expression,and explore the effect of inhibiting FLOT2 expression on proliferation,apoptosis,invasion and migration of colon cancer cells.Methods The expression of FLOT2 in colorectal cancer tissue and LoVo,SW480 and HT29 cells were detected by qRT-PCR,the expression of FLOT2 protein was detected by Western blotting.The expression of FLOT2 was detected by qRT-PCR and Western blotting after the lentivirus vector of FLOT2 siRNA was transfected into LoVo cells and SW480 cells.MTT assay,flow cytometry,Transwell chamber and Western blotting were used to detect the cell proliferation,apoptosis rate,invasion and migration ability and the expression of PCNA,cleaved caspase 3,E-cadherin and vimentin.Results The expression of FLOT2 mRNA and protein in three colon cancer cells was significantly higher than that in NCM460 cells(P<0.05).After FLOT2 siRNA lentiviral vector were transfected into LoVo cells and SW480 cells,the expression of FLOT2 mRNA and protein were significantly reduced,and the ability of cell proliferation,invasion and migration were significantly reduced,apoptosis was promoted,the expression of PCNA and Vimentin was down regulated,and cleaved Caspase3 and E-cadherin were up regulated(P<0.05).Conclusion Inhibition of FLOT2 expression can significantly reduce the proliferation,invasion and migration of colon cancer cells,and promote apoptosis.The mechanism may be related to the regulation of PCNA,cleaved caspase 3,E-cadherin and vimentin expression.

关 键 词：结肠癌 FLOT2基因 增殖 凋亡 侵袭迁移 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]