| ObjectiveChotherapeutic resistance is responsible for recurrence and metastasis of osteosarcoma leading to the failure of treatment.ADD3(γ-Adducin),a member of the adductin family,is involved in the composition of the cell membrane protein network.Meanwhile,the expression of ADD3(γ-Adducin)is increased in tumor cells treated with chemotherapy drugs,which is a potential drug resistance gene.This study aims to investigate the role of down-regulation of ADD3 in adriamycin resistance of osteosarcoma cells MG63.Method(1)After adriamycin induction,the osteosarcoma cell line MG63 was constructed with drug resistance;(2)Real-time quantitative fluorescence PCR(Q-PCR)and Western-blot(WB)techniques were used to detect the differential expression of ADD3 in MG63 cells and MG63/DOX and drug-resistant progeny induced by it.(3)The expression of ADD3 in normal and drug-resistant osteosarcoma cells was downregulated by si RNA to investigate the sensitivity of osteosarcoma cells to chemotherapy drug doxorubicin;(4)The effect of down-regulating ADD3 on adriamycin resistance of osteosarcoma cells was explored through clone formation experiment and flow cytometry.Result(1)Osteosarcoma drug-resistant cell line MG63/DOX was successfully constructed after adriamycin induction.(2)ADD3 was highly expressed in drug-resistant MG63/DOX cells compared with the parental MG63 cells.(3)After the expression of ADD3 silenced,the resistance of MG63 and MG63 / DOX cells to doxorubicin decreases;(4)Treated by adriamycin,down-regulation of ADD3 can increase the apoptosis of osteosarcoma cells and decrease the cell cloning ability.ConclusionDown-regulation of ADD3 can decrease the doxorubicin resistance of osteosarcoma MG63 cells by increasing apoptosis and reducing cell clone formation.. |
PDF Full Text Request