大黄酸通过抑制STAT3基因的表达调控骨肉瘤细胞的增殖和凋亡

目的:探讨大黄酸对骨肉瘤细胞增殖和凋亡的影响及潜在的作用机制。方法:以人骨肉瘤MG-63细胞为研究对象，利用CCK-8实验检测大黄酸对MG-63细胞增殖的影响，并计算大黄酸作用48 h的半数抑制浓度（IC50），采用该浓度进行后续的实验。转染STAT3 siRNA至MG-63细胞，qRT-PCR和Western blot检测分别在mRNA和蛋白水平上检测转染效果，采用CCK-8实验、克隆形成实验、流式细胞术验证大黄酸和STAT3 siRNA及其联合作用对MG-63细胞存活率、克隆形成、凋亡能力的影响，Western blot检测各组MG-63细胞中凋亡相关蛋白Bax、Bcl-2、Cleaved Caspase-3的表达情况。结果:大黄酸能够呈浓度依赖性的抑制骨肉瘤MG-63细胞增殖，干预48 h的IC50为46.05 μmol/L。大黄酸或STAT3 siRNA均能降低MG-63细胞中STAT3的表达，抑制MG-63细胞存活率，阻碍细胞克隆形成能力，促进细胞凋亡，上调细胞中Bax和Cleaved Caspase-3的表达，下调Bcl-2的表达；且二者联合效果更显著。结论:大黄酸通过抑制STAT3基因表达抑制骨肉瘤细胞增殖并促进细胞凋亡。

Rhein regulates proliferation and apoptosis of osteosarcoma cells by inhibiting the expression of STAT3 gene

Objective:To investigate the effect of rhein on proliferation and apoptosis of osteosarcoma cells and its potential mechanism.Methods:Human osteosarcoma MG-63 cells were used as the research object.The effect of rhein on the proliferation of MG-63 cells was detected by CCK-8 assay,and the half-inhibitory concentration(IC50)of rhein for 48 h was calculated.STAT3 siRNA was transfected into MG-63 cells,and transfection was detected by mRNA and protein at qRT-PCR and Western blot,respectively.CCK-8 assay,colony formation assay,flow cytometry were used to verify Rhein and STAT3 siRNA.The effects of the combination on the survival rate,colony formation and apoptosis of MG-63 cells were detected by Western blot.The expressions of apoptosis-related proteins Bax,Bcl-2 and Cleaved Caspase-3 were detected by Western blot.Results:Rhein could inhibit the proliferation of osteosarcoma MG-63 cells in a concentration-dependent manner,and the IC50 of intervention for 48 h was 46.05μmol/L.Rhein or STAT3 siRNA can decrease the expression of STAT3 in MG-63 cells,inhibit the survival rate of MG-63 cells,inhibit the ability of cell clone formation,promote apoptosis,up-regulate the expression of Bax and Cleaved Caspase-3,and down-regulate the expression of Bcl-2,and the combined effect of the two is more significant.Conclusion:Rhein inhibits the proliferation of osteosarcoma cells and promotes apoptosis by inhibiting STAT3 gene expression.