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目的观察RegⅣ基因对5-FU干预结直肠癌LoVo细胞增殖、凋亡的影响。方法构建重组质粒pcDNA3.1-RegⅣ并转染LoVo细胞,RT-PCR和细胞免疫组化检测转染前后细胞中RegⅣ基因的mRNA和蛋白表达。终浓度80μmol/L的5-FU干预细胞48h后,MTT检测细胞的增殖活性。平板克隆实验观察细胞的克隆形成能力。FCM检测细胞凋亡。结果LoVo细胞不表达RegⅣ基因,转染后的LoVo/RegⅣ细胞高表达RegⅣ。5-FU干预后,未转染组(LoVo)和空转质粒组(LoVo/空载)细胞增殖活性、克隆形成能力较RegⅣ转染组(LoVo/RegⅣ)明显降低(P<0.05),细胞凋亡率较RegⅣ转染组明显升高(P<0.05)。结论RegⅣ基因表达可能降低5-FU抑制LoVo细胞的增殖活性及克隆形成能力,并能抑制5-FU诱导LoVo细胞凋亡的作用。RegⅣ基因可能是患者对5-FU化疗产生抗药性的标志及导致临床化疗失败的原因之一。
Objective To investigate the effects of RegⅣ gene on the proliferation and apoptosis of colorectal cancer LoVo cells treated with 5-FU. Methods Recombinant plasmid pcDNA3.1-RegⅣ was constructed and transfected into LoVo cells. The mRNA and protein expression of RegⅣ gene were detected by RT-PCR and immunohistochemistry. After the cells were treated with 5-FU at a concentration of 80μmol / L for 48h, MTT assay was used to detect the proliferation activity. Plate cloning experiments were observed cell clonality. FCM detection of apoptosis. Results The LoVo cells did not express RegⅣ gene, and the LoVo / RegⅣ cells after transfection showed high expression of RegⅣ. After 5-FU intervention, the cell proliferation activity and the ability of colony formation in LoVo and LoVo / Loaded groups were significantly lower than those in LoVo / RegIV group (P <0.05) The mortality rate was significantly higher than that in the RegⅣ-transfected group (P <0.05). Conclusions Reg Ⅳ gene expression may reduce the inhibitory effect of 5-FU on the proliferation and colony-forming ability of LoVo cells and inhibit the apoptosis of LoVo cells induced by 5-FU. RegⅣ gene may be one of the reasons for the emergence of drug resistance to 5-FU chemotherapy and the failure of clinical chemotherapy.