【摘要】 目的探讨热休克蛋白(HSP)27在多发性骨髓瘤中的生物学效应及其作用机制。方法选取2018年1月～2019年12月如皋市人民医院收治的多发性骨髓瘤患者95例。治疗前后取空腹静脉血4 ml分离血清血浆,采用酶联免疫吸附法检测血浆HSP27水平和血清Toll样家族受体-4(TLR4)、核因子κB(NF-κB)水平。慢病毒转染技术在多发性骨髓瘤细胞株RPMI 8226细胞中过表达HSP27,小干扰RNA技术抑制多发性骨髓瘤细胞株RPMI 8226细胞内的HSP27表达,RT-PCR和Western blot法检测HSP27过表达、抑制前后多发性骨髓瘤细胞株RPMI 8226细胞内的HSP27、TLR4和NF-κB表达水平,并同期采用Annexin V-APC/PI双染后流式细胞术检测细胞凋亡率。结果与治疗前比较,治疗后患者血浆HSP27水平、血清TLR4、NF-κB水平均降低,差异有统计学意义(P <0.05)。与HSP27过表达前比较,HSP27过表达后多发性骨髓瘤细胞株RPMI 8226细胞内的HSP27、TLR4和NF-κB的mRNA和蛋白表达水平均升高,细胞凋亡比例降低,差异有统计学意义(P <0.05)。与HSP27抑制前比较,HSP27抑制后多发性骨髓瘤细胞株RPMI 8226细胞内的HSP27、TLR4和NF-κB的蛋白表达水平均降低,细胞凋亡比例升高,差异有统计学意义(P <0.05)。结论 HSP27可抑制多发性骨髓瘤细胞凋亡,调控TLR4和NF-κB表达为其可能作用机制。更多还原

【Abstract】 Objective To investigate the biological effect of HSP27 in multiple myeloma and its mechanism.Methods 95 patients with multiple myeloma admitted to our hospital from January 2018 to December 2019 were selected as the research subjects.Before and after treatment,4 ml of fasting venous blood was taken to separate serum plasma.Enzyme-linked immunosorbent assay was used to detect plasma HSP27 levels,serum TLR4 and NF-κB levels.Lentiviral transfection technology was applied in overexpressing of HSP27 in multiple myeloma cell line RPMI 8226 cells,and small interfering RNA technology was applied in inhibiting HSP27 expression in multiple myeloma cell line RPMI 8226 cells.RT-PCR and Western blotting methods were used to detect the expression levels of HSP27,TLR4 and NF-κB in the RPMI 8226 cells before and after HSP27 overexpression and inhibition.And the flow cytometry after Annexin V-APC/PI double staining were used to detect the multiple myeloma cell line RPMI 8226 cells apoptosis rate before and after HSP27 overexpression and inhibition.Results Compared with those of before treatment,after treatment plasma HSP27 level and serum TLR4 and NF-κB levels of patients decreased(P < 0.05).Compared with those of before HSP27 overexpression,the after HSP27 overexpression mRNA and protein expressions of HSP27,TLR4 and NF-κB in multiple myeloma cell line RPMI 8226 cells increased,and the after HSP27 overexpression multiple myeloma cell line RPMI 8226 cells percentage of apoptosis decreased(P < 0.05).Compared with those of before HSP27 inhibition,the after HSP27 inhibition protein expression levels of HSP27,TLR4 and NF-κB in multiple myeloma cell line RPMI 8226 cells decreased,and the after HSP27 inhibition multiple myeloma cell line RPMI 8226 cells percentage of apoptosis increased(P < 0.05).Conclusion HSP27 can inhibit the apoptosis of multiple myeloma cells,and its possible mechanism is to regulate the expression of TLR4 and NF-κB.更多还原