【摘要】 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。更多还原

【Abstract】 Objective To study the effect of Reg Ⅳ on 5-Fluorouracil(5-FU) intervened human colorectal carcinoma cell line LoVo proliferation and apoptosis.Methods Recombined plasmid pcDNA3.1-Reg Ⅳ was constructed and then transfected into LoVo cell. The Reg Ⅳ mRNA and protein levels in cells were confirmed by RT-PCR and immunohistochemistry.After the cells were intervened by 5-FU for 48 hours,cell proliferating potency and colony-forming ability were detected by MTT assay and flat plate colony forming experiment separately.Cell apoptosis was measured by flow cytometry.Results There was no Reg Ⅳ expression in untreated LoVo cells but highly expressed Reg Ⅳ in Reg Ⅳ transfected cells. After the cells were intervened by 5-FU,the LoVo cell proliferation and colony-forming ability of the blank control group and the no-load control group were significantly lower than those of experimental group(P<0.05). The apoptosis rate of blank control group and the no-load control group was significantly higher than that of experimental group(P<0.05).ConclusionReg Ⅳ could reduce the effect of 5-FU on LoVo cell proliferation and colony-forming ability,and inhibit LoVo cell apoptosis induced by 5-FU. Reg Ⅳ may be a drug resistance marker of 5-FU chemotherapy and one of the areasons for the failure of clinical chemotherapy.更多还原