脂质体槲皮素抑制热休克后人肝癌HEPG-2和肾癌ORSC-2细胞系HSP70表达

Inhibition of the expression of HSP70 in HEPG-2 and ORSC-2 cell lines by liposome quercetin after hyperthermia

目的比较不同浓度脂质体槲皮素(LQ)对人肝癌HEPG-2细胞和肾癌ORSC-2细胞热疗后热休克蛋白70的表达变化及其作用机制。方法用旋转蒸发法制备LQ。将细胞分为37℃培养组、42℃培养组和槲皮素干预组。MTT法检测细胞抑制率,优选出合适浓度。用10、15和30 mg/L的LQ处理上述两种肿瘤细胞,于热休克后2、4、6、8和10 h取出细胞。流式细胞计量术检测细胞凋亡,RT-qPCR检测HSP70mRNA的表达,Western blot检测细胞HSP70的表达水平。结果42℃热疗后人肝癌HEPG-2细胞和肾癌ORSC-2细胞的HSP70在基因和蛋白水平上均明显升高(P<0.05)。随着脂质体槲皮素浓度的增加,HSP70表达逐渐降低。结论脂质体槲皮素能通过抑制热休克状态肿瘤细胞HSP70的表达诱导肝癌和肾癌肿瘤细胞凋亡。

Objective To investigate the effects and mechanisms of the expression of heat shock protein 70(HSP70)by human hepatocellular carcinoma HEPG-2 cells and renal cell carcinoma ORSC-2 by different concentrations of liposomal quercetin(LQ)after hyperthermia.Methods The method of rotary evaporation was used to prepare quercetin.The cells were divided into 37℃control group,42℃hyperthermia group and quercetin intervention group.Two kinds of tumor cells were treated with LQ at 10,15 and 30 mg/L.The tumer cells were incubated at 42℃for 2 h,and cells were removed at 2,4,6,and 10 h after hyperthermia.Flow cytometry analysis was applied to determine the apoptosis of tumer cells,and the change of mRNA and protein level of HSP70 was detected by real-time PCR and Western blot respectively.Results After hyperthermia,the expressions of HSP70 in HEPG-2 cells and ORSC-2 cells were significantly increased at both gene and protein levels(P<0.05).With the increase of the concentration of LQ,the expression of HSP70 gradually decreased.Conclusions Liposome quercetin can induce apoptosis of hepatocellular carcinoma and renal carcinoma cells by inhibiting the expression of HSP70 in tumor cells after hyperthermia.