摘要
目的建立卵巢癌顺铂(cisplatin,DDP)耐药细胞系,并比较耐药与非耐药卵巢癌细胞系之间部分耐药相关基因的表达差异。方法采用浓度梯度递增法建立人卵巢癌DDP耐药细胞系SKOV-3/DDP。验证其有关的生物学指标;并用免疫组化法及RT-PCR技术检测耐药与非耐药卵巢癌细胞系之间P-糖蛋白(P-glycoprotein,P-gp)及部分耐药相关基因的表达差异。结果耐药细胞系对DDP、氟尿嘧啶(fluorouracil)和多柔比星(doxorubicin)的耐药指数(RI)分别为13.94、10.83、4.52;细胞周期比例SKOV-3/DDP细胞系G0/G1期细胞明显少于SKOV-3细胞系(P<0.01),S期、G2/M期细胞明显多于SKOV-3细胞系(P<0.01);P-糖蛋白(P-gp)阳性细胞率SKOV-3/DDP细胞系为65%±5%;SKOV-3细胞系为6%±2%,两者差异极显著(P<0.001)。在检测的耐药相关基因中MDR1、Sur-vivin在SKOV-3/DDP细胞系呈阳性表达,在SKOV-3细胞系未表达;TopoⅡβ表达情况与此相反。结论顺铂诱导的卵巢癌细胞耐药可对其他不同作用机制的药物产生交叉耐药,多药耐药涉及多个相关基因的表达,说明耐药的发生是一个多途径的复杂过程。
Objective To establish DDP - resistant ovarian cancer cell strain , and investgate expression difference of several drug resistance related genes. Methods Cell line resistant to cisplatin was established from ovarian cancer cell line of SKOV - 3 and their biological features were determined. The expressions of several genes associated with drug resistance and P - glycoprotein were measured by RT - PCR and immunohistochemistry. Results The resistance index （RI） of SKOV - 3/DDP cell to DDP, fluorouracil , doxorubicin was 13.94, 10.83 and 4.52; G0/G1 of SKOV - 3/DDP cell line was less than that of SKOV - 3 cell line（ P 〈 0.01）, while S,G2/M of SKOV - 3/DDP cell line were more than that of SKOV - 3 cell line（ P 〈 0.01）. P-glycoprotein positive ratio of SKOV - 3/DDP cell line was 65% - 5% ,while that of SKOV - 3 cell line was 6% ± 2% （ P 〈 0.001 ）. MDR1 .Survivin was positive in SKOV - 3/DDP cell line and was negative in SKOV - 3/DDP cell line, Topo Ⅱβ was negative in SKOV - 3/DDP cell line and was positive in SKOV - 3/DDP cell line. Conclusion Induction of cisplatin resistance in ovarian cancer may lead cross resistance of other drugs which are in different mechanism working on cancer cells, multiple drug - resistance involve several related genes, suggesting more than one pathway leading to resistance transformation.
出处
《中国热带医学》
CAS
2006年第12期2132-2134,共3页
China Tropical Medicine
关键词
卵巢癌
顺铂
多药耐药
基因表达
Ovarian cancer
Cisplatin
Multiple drug resistance
Gene expression
