硼替佐米增强P16蛋白表达诱导骨髓瘤细胞衰老

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。

Bortezomib Induced Myeloma Cells Senescence by Enhancing the Expression of P16 Protein

Objective： To study the proteasome inhibitor bortezomib induced myeloma RPMI8226,MMH929 cell senescence,and further explore its mechanism.Methods： Myeloma RPMI8226,MMH929 cell were routinely cultured and treated with different concentrations of bortezomib（1-500 nmol/L） for 48-72 h.Viability was determined by MTT assay.The drug of choice IC50 values of 1/10 dose treatment RPMI8226,MMH929 cell 0,24,48 H.Apply PI staining method to test cell cycle by flow cytometry.Apply SA-β galactosidase staining method to test the senescence of RPMI8226,MMH929 cell induced by bortezomib.The protein levels of P53,PTEN,P16 were determined by Western bolt.Results： Bortezomib treatment of myeloma cells RPMI8226,H929 48-hour IC50 values： RPMI8226： 19.05nmol / L,H929： 18.45nmol / L.Treatment with Bortezomib 2 nmol / L after 0,24,48 H,indicating β-galactosidase staining rate,proportion of cells G0/G1 is up-regulated.The protein level of P16 was up-regulated while that of P53,PTEN did not change by Western blot.Conclusion： Bortezomib induced myeloma cells RPMI8226,H929 senescence by enhancing the expression of P16 protein.