摘要
目的 探讨和研究热休克蛋白 90 (hsp90 β)与肿瘤恶性表型和耐药性的关系。 方法 将hsp90 β正义、反义表达质粒及pLXSN空载体 ,lipofectin法转染HeLa细胞。G418抗性筛选。台盼蓝染色法 ,绘细胞生长图。MTT法检测细胞对药物的敏感性。软琼脂细胞集落形成实验检测肿瘤细胞恶性表型。结果 转染正义hsp90 β表达质粒的细胞生长与转染空载体细胞及HeLa细胞差别不大。而转染hsp90 β反义表达质粒的细胞生长慢于对照组细胞 ,对常用的化疗药物阿霉素 (ADM)、顺铂(CDDP)的药敏感性比对照组细胞明显升高 ,其半致死剂量仅为后两者的 1/2和 1/5 ;软琼脂集落实验表明 ,转染反义hsp90 β表达质粒的细胞集落形成能力减弱。 结论 反义hsp90 β在一定程度上抑制了HeLa细胞的生长 ,降低HeLa细胞恶性表型 ,明显增强HeLa细胞对化疗药物的敏感性。
Objective To study the effects of hsp90β on tumorigenesis and sensitivity of HeLa cells to chemotherapeutic drugs. Methods A recombinant plasmid expressing either sense or antisense hsp90β RNA, and a vector pLXSN were transfected into HeLa cell line by lipofectin method. The transfected cells were screened by G418. The growth rates of the transfected HeLa cells and their sensitivity to adriamycin(ADM) and cisplatin(CDDP) were examined by cell count and MTT uptake. Colony formation in soft agar was used to embody the malignant phenotype in vitro. Results There was little difference between HeLa cells transfected with sense RNA plasmids and pLXSN vector. The growth rate of HeLa cells transfected with antisense RNA declined, and their ability to form colony in soft agar was decreased. The sensitity of the antisense hsp90β transfected HeLa cells to ADM and CDDP increased 2.1～5.8 fold; the LD 50 decreased to 1/2～1/5 that of the wild type HeLa cells.Conclusion Hsp90β is involved in the maintenance of malignant phenotype and drug resistance in HeLa cells.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2000年第1期14-16,共3页
Chinese Journal of Oncology
基金
国家自然科学基金! ( 3 9770 82 4)
中国医学科学院基金! ( 95 2 0 11)
