GRP78对SMMC-7721细胞耐药性的影响及机制  

作　　者：蔡成行[1] 李光明[1] 雷康[1] 黄长文[1] 傅华群[1] 

机构地区：[1]南昌大学第二附属医院肝胆外科

出　　处：《当代医学》2009年第24期76-78,共3页Contemporary Medicine

摘　　要：目的探讨GRP78对人肝癌细胞株SMMC7721细胞耐药性和Topo-Ⅱ表达的影响。方法用低浓度顺铂(cDDP)短期诱导SMMC-7721细胞产生耐药,并用脂质体转染不同浓度GRP78反义寡核苷酸(GRP78ASODN)以降低SMMC-7721细胞GRP78的表达,应用RT-PCR检测并筛选一个最有效的降低GRP78mRNA表达的GRP78ASODN浓度,然后用MTT法检测亲本SMMC-7721细胞、耐药细胞及筛选出的转染组细胞对cDDP的药物敏感性,得出各组的半数抑制浓度(IC50)及耐药指数,并用RT-PCR检测三组细胞Topo-ⅡmRNA的表达。结果RT-PCR显示转染GRP78ASODN浓度为100nmol/ml时,GRP78mRNA降低最为显著(P<0.01);耐药组Topo-ⅡmRNA的表达较亲本细胞相显著降低(P<0.01),转染组与耐药组相比显著升高(P<0.01)。经诱导得到的耐药细胞,IC50是亲本细胞IC50的1.91倍,转染后,其IC50与转染前相比其耐药性显著降低(P<0.01)。结论用低浓度cDDP短期诱导SMMC-7721细胞产生耐药;GRP78能提高SMMC-7721细胞的耐药性,并与Topo-Ⅱ有关;抑制GRP78可以降低SMMC-7721细胞的耐药性。Objective To observe the impact of GRP78 on SMMC-7721 resistance and Topo-Ⅱ expression. Methods using low concentrations of cisplatin （cDDP） short term inducting to produce resistance and transfect GRP78ASODN into SMMC-7721 to reduce the expression of GRP78mRNA,the expression of GRP78mRNA was detected by R-T-PCR . screening one of the most effective concentration. Drug sensitivity of parental SMMC-7721 cells,drug-sesistant cells and the transrected cells was detected by MTT assay, obtained IC50 of each group as well as resistance index. Topo-Ⅱ mRNA expression of each group was detected by RT-PCR. Results RT-PCR showed that when transfected 100nmol/ml GRP78ASODN, GRP78mRNA reduced to the most signifi-cant （P〈0.01）. Expression of Topo-Ⅱ mRNA in drug-resistant group was significantly lower （P〈0.01） than parental cells. But transfection group was significantly higher than resistant group （P〈0.01）. The IC50 of drug-resistant cells are significantly higher than parental cells, （P〈0.01）, after transfection its IC50 decreas significantly （P〈0.01）. Conclusion The short low concentrations of CDDP-induced SMMC-7721 cells to produce resistance; GRP78 can enhance SMMC-7721 cells＇ resistance, and relate with Topo-Ⅱ; Inhibitting GRP78 can reduce the SMMC-7721 cells＇ resistant.

关 键 词：葡萄糖调节蛋白 癌 肝细胞 拓扑异构酶Ⅱ 抗药性 

分 类 号：R735.7[医药卫生—肿瘤] R730.53[医药卫生—临床医学]