胶质瘤多药耐药细胞系C6/adr的建立及其生物学特性的研究

目的　为研究脑胶质瘤的耐药、机理及逆转,建立了C

Establishment of a multidrug resistant C6/adr cell line and its characterization

Objective　To study drug resistant mechanism and reversal of glioma we established C6 MDR drug-resistant cell line(C6/adr). Methods　This cell line was obtained by sucessive exposure to increasing amount of adriamycin and associated with high dose short explosure of ADM.A series of biological characteristics of C6 adr were examined, The morphology and microstructure were studied with HE and electron microscope, cell line growth characteristics were studied by MTT. The distribution of cell cycle, the content of DNA and RNA were analysed by FCM. Clone forming ratio and chromosome were observed. Multidrug resistance gene mdr-1 and its encoded P-gp were tested quantitatively and qualitatively though RT-PCR, immunocytochemical technique and FCM. The telomerase was also examined by RT-PCR. C6/adr SD models were established successfully and their drug resistance was analysed. Meanwhile the sensitivity test of C6/adr to anticancer drugs were performed with MTT method. Results　C6/adr was the same as C6 on the morphology(HE), but by transmission electron microscope, there were lots of malformed nuclei, abundant mitochondria and ribosomes, endoplasmic reticular. Populaion doubling time was 55.2 hrs, After C6/adr generating 2,3 days, DNA and RNA index had no changes; Cell cycle distribution was major in G0＋G1,clone forming rate was 55%; Chromosomes were on No.40 to 42, most in subdiploid, P-gp(42.5±7.82%) and telomerase expressed positive strongly. No changes in morphology (HE) and content of P- gp were observed in SD rat model.Drug resistance was to ADM,VM-26,VCR,5F-r,VP-16,CDDP,no drug resistance to MTX,CCNU.Conclusions　 C6/adr cell line encoded drug resistance gene is MDR-1. This results show that this cell line may be useful in further studying on drug-resistance and reversan of glioma.