苦参碱对人骨肉瘤MG63细胞增殖、凋亡的影响及机制

Effects of matrine on proliferation and apoptosis of human osteosarcoma MG63 cells and its mechanism

目的探讨苦参碱对人骨肉瘤MG63细胞增殖、凋亡的影响及机制。方法将人骨肉瘤MG63细胞分为空白对照组和低、中、高剂量实验组,空白对照组细胞不做任何处理,低、中、高剂量实验组分别加入0. 5,1. 0,1. 5 mg·mL^(-1)苦参碱。用甲基偶氮唑蓝比色法(MTT)检测MG63细胞增殖情况,Annexin VFITC/PI双染流式细胞术检测MG63细胞凋亡情况,蛋白免疫印迹法检测MG63细胞中基质金属蛋白酶(MMPs)蛋白表达情况。结果低、中、高剂量实验组细胞24 h增殖抑制率分别为(16. 66±2. 18)%,(26. 84±2. 48)%,(31. 46±3. 62)%,差异均有统计学意义(P <0. 05或P <0. 01);细胞凋亡率分别为(3. 47±0. 54)%,(4. 75±0. 57)%,(5. 53±0. 68)%,与空白对照组的(2. 58±0. 74)%相比,差异均有统计学意义(P <0. 05或P <0. 01)。空白对照组、低、中、高剂量实验组中,MMP-2蛋白相对表达量分别为1. 61±0. 52,1. 27±0. 35,0. 86±0. 22,0. 63±0. 12,MMP-9蛋白相对表达量为2. 35±0. 43,1. 98±0. 32,1. 21±0. 23,0. 89±0. 14,差异均有统计学意义(均P <0. 01)。结论苦参碱可抑制人骨肉瘤MG63细胞增殖,促其凋亡,并降低MMPs相关蛋白水平。

Objective To investigate the effect of matrine on the proliferation and apoptosis of human osteosarcoma MG63 cells and its underlying mechanism. Methods Human osteosarcoma MG63 cells were divided into blank control group and experimental-L/-M/-H group.The experimental-L/-M/-H group were treated with 0. 5,1. 0 and 1. 5 mg ·mL^-1 of matrine,while the blank control was treated without matrine. The proliferation rates of MG63 cells was detected by methylthiazolyldiphenyl-tetrazolium( MTT),the apoptosis rates of MG63 cells was detected by Annexin V-FITC/PI double staining flow cytometry. Western blotting( WB) was used to detect the expression of matrix metalloproteinases( MMPs) in MG63 cells. Results There were significant differences among the experimental-L group,-M group and-H group in the 24 h proliferation inhibition rates [( 16. 66 ± 2. 18) %vs( 26. 84 ± 2. 48) % vs( 31. 46 ± 3. 62) %]. There were significant differences between the blank control group and the experimental-L/-M/-H group in the apoptotic rates [( 2. 58 ± 0. 74) % vs( 3. 47 ±0. 54) %,( 4. 75 ± 0. 57) %,( 5. 53 ± 0. 68) % ]( P < 0. 05 orP < 0. 01). There were significant differences between the blank control group and the experimental-L/-M/-H group in the protein expressions of MMP-2 [1. 61 ± 0. 52 vs 1. 27 ± 0. 35,0. 86 ± 0. 22,0. 63 ± 0. 12] and MMP-9[2. 35 ± 0. 43 vs 1. 98 ± 0. 32,1. 21 ± 0. 23,0. 89 ± 0. 14]. Conclusion Matrine can inhibit the proliferation and promote apoptosis of human osteosarcoma MG63 cells,and reduce the level of MMPs related proteins.