摘要
目的:利用5-氟尿嘧啶-MKN45胃癌细胞系模型,研究p53β异构体表达的生物学意义。方法:不同浓度5-氟尿嘧啶(5-fluorouracil,5-FU)作用于MKN45胃癌细胞系,MTT法检测细胞抑制率;巢式逆转录多聚酶链反应(nested Reverse transcription-polymerase chain reaction,nRT-PCR)检测p53β、p53和Bax mRNA的表达变化。Pearson直线分析观察p53β表达水平与p53及Bax表达水平的相关性。结果:MTT检测结果显示,5-FU对MKN45的生长有显著的增殖抑制作用,且呈剂量和时间双效应,浓度为25、50和100μg/mL的5-FU作用于MKN45细胞48h后,抑制率分别为37.23%、59.54%和77.95%,差异有统计学意义,H=17.857,P<0.01;浓度为25μg/mL的5-FU作用于MKN45细胞24、48和72h后,抑制率分别为17.43%、37.23%和78.09%,差异有统计学意义,F=19.108,P<0.01。RT-PCR结果显示,MKN45细胞中p53β、p53和Bax mRNA表达均随5-FU浓度增加而增加,H值分别为15.118、19.785和21.963,P值均<0.01。Pearson直线相关分析结果显示,p53β表达水平与p53表达水平呈显著正相关,r=0.976,P<0.05;与Bax表达水平呈显著正相关,r=0.994,P<0.01。结论:p53β异构体表现出与野生型p53协同作用于下游分子Bax的生物学功能,提示p53β异构体在p53诱导的肿瘤生长抑制效应中发挥重要作用。
OBJECTIVE：To study the expression of p53β and its biological significance in 5-fluorouracil-human gas- tric cancer cell line MKN45 model. METHODS： MKN45 gastric cancer ceils were interefered with various concentrations of 5-flurouracil （5-FU）,and MTT assay was applied to detect the growth inhibition rates, p53β,p53 and Bax mRNA were detected by nested Reverse transcription-polymerase chain reaction （nRT-PCR）. The relevence between the expression of p53β and wide-type p53 or gax was shown by Pearson linear analysis. RESULTS： In MTT assay,it was shown that the MKN45 gastric cancer cells were significantly inhibited by 5-FU. The growth inhibition rates were positively related to both concentration and culturing time,the inhibition rates of 25,50 and 100 μg/mL were 37.23%,59.54%,77. 95%,se- perately,for culturing 48 hours. The difference was significant among groups with various 5-FU concentration （H= 17. 857, P〈0.01 ）. The inhibition rates of 25 μg/mL were 17.43%, 37.23 %, 78.09%, seperately, for culturing 24,48 and 72 hours. The difference was significant among groups with different culturing time （F=19. 108 ,P〈0.01）. In RT-PCR analysis, the mRNA levels of p5313, p53, and Bax increased with the growing of 5-FU concentrations, and the difference was significant （H= 15.118,19. 785,21. 963, P〈I0.01, respectively）. Pearson linear correlation analysis showed that the ex- pression level of p53β was positively related to that of p53 （r =0. 976, P 〈 0.05 ）, and to that of Bax （r = 0. 994, P 0.01） ,and the correlation was significant. CONCLUSION： The p53β isoform playes as a co-operater with wide-type p53 and its downstream molecule-gax, which indicated its key role in p53-induced tumor growth inhibition.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2014年第5期352-355,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
山东省优秀中青年科学家科研奖励基金(BS2010SW034)
