【摘要】 目的观察p14ARF基因转染联合氟尿嘧啶对人结肠癌LOVO细胞的作用。方法采用RT-PCR和Western blot方法检测转染前后LOVO细胞中p14ARF基因的表达。四唑蓝比色法(MTT法)检测细胞增殖,流式细胞术分析细胞周期,对比在不同浓度的氟尿嘧啶作用下转染前后LOVO细胞的增殖情况。结果LOVO细胞呈现p14ARF基因缺失变异。RT-PCR和Western blot法证实转染后LOVO细胞中有p14ARF基因的表达。p14ARF基因转染后细胞增殖受抑,流式细胞仪显示p14ARF基因诱发凋亡。在氟尿嘧啶0.1g/mL的作用下,未转染的LOVO细胞的增殖速度未受影响,而转染后LOVO细胞较未用药时的抑制率增加;氟尿嘧啶1、10、100μg/mL时,对转染前后LOVO细胞均有增殖抑制作用,且呈剂量相关性。氟尿嘧啶对转染后LOVO细胞的抑制率明显高于未转染的LOVO细胞。结论p14ARF基因转染可抑制LOVO细胞增殖。p14ARF基因和氟尿嘧啶联合用于LOVO细胞能够加强氟尿嘧啶的增殖抑制作用,明显提高LOVO细胞对氟尿嘧啶的敏感性。更多还原

【Abstract】 Purpose To investigate the effects of combination of fluorouracil with p14ARF gene transfection on human colon cancer cell line LOVO. Methods Expressions of the p14ARF mRNA and protein in LOVO cells before and after transfection were measured by the RT-PCR and Western blot.Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) colorimetric assay.Cell cycle was analyzed by flow cytometry.Under different concentration of fluorouracil,the proliferation of untransfected LOVO cells and transfected LOVO cells were compared. Results p14ARF gene was deleted in human colon cancer cell line LOVO.The expressions of p14ARF in transfected LOVO cells were detected by the RT-PCR and Western Blot methods.The proliferation of LOVO cells was inhibited after the transfection.The transfection of p14ARF gene induced apoptosis.The proliferation of untransfected LOVO cells was not inhibited when treated with fluorouracil of 0.1 μg/mL.While the proliferation of transfected LOVO cells was inhibited at the same concentration.Both untransfected and transfected LOVO cells was inhibited when treated with fluorouracil of 1,10 and 100 μg/mL respectively.The inhibition degree was correlated with the dose of fluorouracil. The proliferation inhibition of transfected LOVO cells was greater than that of untransfected LOVO cells. Conclusions The transfection of p14ARF gene inhibits proliferation of LOVO cells.The combination of fluorouracil with p14ARF gene could increase the inhibitory function of fluorouracil and sensitize LOVO cells to fluorouracil.更多还原