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作 者:郑洋[1] 潘娅[2] 洪阳 向敏[4] 王寒琪[1] 王燕[2] 陈腾祥[2]
机构地区:[1]贵阳医学院附院消化内科,贵州贵阳550004 [2]贵阳医学院生理学教研室,贵州贵阳550004 [3]贵阳市卫生局,贵州贵阳550001 [4]贵阳医学院附院儿科,贵州贵阳550004
出 处:《贵阳医学院学报》2012年第3期235-237,258,共4页Journal of Guiyang Medical College
基 金:国家自然科学基金(81060176);贵阳市科技局社会发展领域科技攻关项目:筑科农字(2010)
摘 要:目的:探讨干细胞相关基因Wnt5b和Nanog在阿霉素(ADM)作用下的人低转移性肝癌细胞系MH-CC97-L中的表达。方法:MHCC97-L细胞常规培养,用四甲基偶氮唑盐比色法(MTT)检测ADM作用72 h时MHCC97-L的半数致死量(LD50);蛋白质免疫印迹法(Western blot)检测ADM(LD50)作用下,干细胞相关基因Wnt5b和Nanog蛋白表达。结果:MTT实验结果显示,ADM对MHCC97-L的半数致死量(ID50)为0.423 3μmol/L;Western blot检测发现,在ADM(ID50)作用下,随作用时间延长,Wnt5b蛋白表达水平逐渐增高,12 h时达最高峰(P<0.05),以后则逐渐降低,72 h时降到最低(P<0.05);在ADM(ID50)作用下,随作用时间延长,Nanog蛋白表达水平逐渐降低,12 h时降到最低,以后则逐渐升高,72 h时达到最高(P<0.05)。结论:干细胞相关基因Wnt5b、Nanog在ADM作用下的人肝癌细胞系MHCC97-L中均有不同程度的表达。Objective: To explore the expression of stem cell associated gene WntSb and Nanog in hepatocellular carcinoma cell line MHCC97-L incubating with adriamycin (ADM). Methods: MH- CC97-L cells were routinely cultured. Median lethal dose ( LD50 ) of MHCC97-L treated with ADM for 72 h were determined with MrlT method. The expression of WntSb and Nanog proteins in MHCC97-L after treating with ADM ( LDso ) was detected with Western blot. Results: MTY results showed that LDso of MHCC97-L treated with ADM was 0. 423 3 μmol/L. The expression of WntSb increased when MHCC97-L was exposed to ADM within 4 hours, and reached peak at 12 hours, and then decreased gradually, and reduced to minimum at 72 h. When the incubation time was less than 12 h, Nanog de- creased gradually with the time passing(P 〈0. 05), and reduced to minimum at 12 h, and then it in- creased gradually, reached peak at 72 hours. Conclusions: WntSb and Nanog proteins express in MH- CC97-L exposed to ADM in different degrees.
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