p14ARF基因联合氟尿嘧啶对人结肠癌LOVO细胞的作用  

The effects of combination of fluorouracil with p14ARF gene transfection on human colon cancer cell line LOVO

作　　者：张文[1] 罗建民[2] 欧周罗[2] 陆洪芬[3] 洪小南[1] 许立功[1] 

机构地区：[1]复旦大学附属肿瘤医院化疗科,上海200032 [2]复旦大学附属肿瘤医院中心实验室,上海200032 [3]复旦大学附属肿瘤医院病理科,上海200032

出　　处：《复旦学报（医学版）》2006年第1期63-67,73,共6页Fudan University Journal of Medical Sciences

基　　金：复旦大学附属肿瘤医院院级基金资助

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。Purpose To investigate the effects of combination of fluorouraeil with p14ARF gene transfeetion on human colon cancer cell line LOVO. Methods Expressions of the p14ARF mRNA and protein in LOVO cells before and after transfeetion 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） eolorimetrie assay. Cell cycle was analyzed by flow eytometry. Under different concentration of fluorouraeil, the proliferation of untransfeeted LOVO cells and transfeeted LOVO cells were compared. Results p14ARF gene was deleted in human colon cancer cell line LOVO. The expressions of p14ARF in transfeeted LOVO cells were detected by the RT-PCR and Western Blot methods. The proliferation of LOVO cells was inhibited after the transfeetion. The transfeetion of p14ARF gene induced apoptosis. The proliferation of untransfeeted 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 pl4ARF gene inhibits proliferation of LOVO cells. The combination of fluorouracil with pl4ARF gene could increase the inhibitory function of fluorouracil and sensitize LOVO cells to fluorouracil.

关 键 词：结直肠癌 化疗 氟尿嘧啶 P14ARF基因 

分 类 号：R735.35[医药卫生—肿瘤]