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目的:研究蛋白酶体抑制剂硼替佐米诱导骨髓瘤RPMI8226、MMH929细胞衰老作用,并进一步探讨其作用机制。方法:硼替佐米0.1-100nmol/L处理骨髓瘤RPMI8226、MMH929细胞48、72h,MTT法检测细胞存活率、药物IC50值。选择药物IC50值1/10剂量处理骨髓瘤RPMI8226、MMH929细胞0、24、48H后检测衰老相关β-半乳糖苷酶染色率。流式细胞术检测细胞周期情况及凋亡率。Western-blot检测相关蛋白表达。结果:硼替佐米处理骨髓瘤细胞RPMI8226、MMH929后48小时IC50值:RPMI8226:19.05 nmol/L,MMH929:18.45nmol/L。以硼替佐米2 nmol/L处理骨髓瘤RPMI8226、MMH929细胞0、24、48H后发现β-半乳糖苷酶染色率、细胞G0/G1期比例明显上升与药物作用时间呈正相关,Western-blot检测细胞周期调控蛋白发现P53、PTEN蛋白无变化,P16蛋白与药物作用时间正相关。结论:硼替佐米通过增强P16蛋白表达诱导骨髓瘤细胞RPMI8226、H929衰老。
OBJECTIVE: To study the senescence effect of proteasome inhibitor bortezomib on myeloma RPMI8226 and MMH929 cells and to explore its mechanism. Methods: Myeloma RPMI8226 and MMH929 cells were treated with bortezomib 0.1-100 nmol / L for 48 and 72 h respectively. Cell viability and drug IC50 were measured by MTT assay. Select the drug IC50 1/10 dose treatment of myeloma RPMI8226, MMH929 cells 0,24,48 H after aging-related β-galactosidase staining rate. Flow cytometry was used to detect cell cycle and apoptosis rate. Western-blot detection of related protein expression. Results: IC50 values of RPMI8226 and MMH929 after treatment with bortezomib were 48h: RPMI8226: 19.05 nmol / L and MMH929: 18.45 nmol / L. The percentage of β-galactosidase staining in MMH929 cells treated with bortezomib 2 nmol / L was significantly higher than that of MMH929 cells. The proportion of cells in G0 / G1 phase was positively correlated with the time of drug treatment. Western-blot assay Cell cycle regulatory protein found P53, PTEN protein no change, P16 protein and drug action time is positively correlated. Conclusion: Bortezomib can induce the senescence of myeloma RPMI8226 and H929 cells by increasing the expression of P16 protein.