硫化氢抑制骨肉瘤U2OS细胞的体外生长  

Hydrogen Sulfide Inhibits Growth of Osteosarcoma Cells U2OS In Vitro

作　　者：郭海华[1] 李敬春 冯鉴强[3] 张泷涓[4] 雍碧城[1] 黄纲[1] 

机构地区：[1]中山大学附属第一医院骨肿瘤科,广东广州510080 [2]广州市妇女儿童医疗中心骨科,广东广州510623 [3]中山大学中医学院生理教研室,广东广州510080 [4]中山大学附属第一医院外科实验室,广东广州510080

出　　处：《中山大学学报（医学科学版）》2013年第3期358-363,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省科技计划项目(2012B031800050)

摘　　要：【目的】探讨硫化氢单独及与顺铂联用对骨肉瘤细胞生长的影响及其可能机制。【方法】分别用0.2、0.4、0.6、0.8、1.0、2.0、3.0、4.0、5.0 mmol/L NaHS(硫化氢载体)单独或与20 mg/L顺铂共同作用骨肉瘤U2OS细胞24 h,CCK-8比色法法检测细胞存活率;分别用0.4及1.0 mmol/L NaHS作用骨肉瘤U2OS细胞24 h,1.0 mmol/L NaHS单独或与20 mg/L顺铂联用作用骨肉瘤U2OS细胞24 h,细胞单克隆形成法检测各组单克隆数;1.0 mmol/L NaHS与20 mg/L顺铂共同作用骨肉瘤U2OS细胞24 h,Western-Blot法检测Bcl-2、Bax、NFκB蛋白的表达。【结果】硫化氢对U2OS细胞生长的影响:0.2、0.4、0.6、0.8 mmol/L NaHS作用U2OS细胞24 h,细胞存活率与对照组比较,差异无统计学意义(P>0.05);1、2、3、4、5 mmol/L NaHS作用U2OS细胞24 h,细胞存活率分别为(90.01±7.49)%、(88.00±4.12)%、(87.26±4.05)%、(85.40±4.39)%和(82.99±4.65)%,与对照组比较,能不同程度抑制U2OS细胞的生长(P<0.05)。0.4 mmol/L NaHS组、1 mmol/L NaHS组细胞单克隆数分别为445.00±25.00、306.00±17.69,与对照组(464.33±8.32)比较,1 mmol/L NaHS组细胞单克隆数减少(P<0.01);0.4mmol/L NaHS组差异无统计学意义(P>0.05)。NaHS与顺铂联用对骨肉瘤细胞的影响:分别用0.8、1、2、3、4、5 mmol/L NaHS联合20 mg/L顺铂共同作用U2OS细胞24 h,细胞存活率分别为(54.13±4.03)%、(54.09±1.13)%、(50.37±4.56)%、(41.44±3.95)%、(40.00±4.34)%和(36.35±3.90)%,与单用顺铂组(59.76±3.15%)比较,存活率下降(P<0.05)。NaHS+顺铂组细胞单克隆数为231.00±13.52,与顺铂组(285.67±11.59)比较,细胞单克隆数下降(P<0.01);NaHS+顺铂组NFκB蛋白表达及Bcl-2/Bax蛋白表达比值较顺铂组下降。【结论】1 mmol/L浓度以上的NaHS能抑制骨肉瘤细胞的生长,0.8 mol/L浓度以上的NaHS通过减少NFκB入核,增加细胞的凋亡,从而增强顺铂对骨肉瘤细胞的抑制作用。[Objective] To investigate the effect of hydrogen sulfide （H2S） alone or combined with cisplatin on osteosarcoma cells U2OS and the possible mechanism in vitro.[Method] Osteosarcoma cells U2OS were treated with NaHS （a donor of H2S） at different concentration （0.2,0.4,0.6,0.8,1.0,2.0,3.0,4.0,and 5.0 mmol/L） alone or combined with cisplatin at 20 mg/L concentration for 24 h respectively,then cell viability was tested by cell counter kit-8.U2OS cells were treated with NaHS at 0.4,and 1.0 mmol/L concentration for 24 h respectively,and U2OS cells were treated with NaHS at 1.0 mmol/L concentration alone or combined with cisplatin at 20 mg/L concentration for 24 h,then the number of monoclone was counted by monoclonal formation experiment.U2OS cells were treated with NaHS at 1.0 mmol/L concentration combined with cisplatin at 20 mg/L concentration for 24 h,then the expressions of Bcl-2,Bax,and NFκB were evaluated by Western blot assay.[Result] The effects of hydrogen sulfide on U2OS cells：At the concentration from 1 to 5 mmol/L,NaHS dose-dependently inhibited cell viability in U2OS cells.Compared with control group,the monoclonal number of 1 mmol/L NaHS group was declined significantly （P 〈 0.05）.The effect of hydrogen sulfide combined with cisplatin on U2OS cells：U2OS cell viability was dose-dependently declined by NaHS at the concentration from 0.8 to 5 mmol/L combined with cisplatin at 20 mg/L concentration for 24 h.Compared with cisplatin group,the monoclonal number of NaHS＋ cisplatin group was declined （P 〈 0.01）.The expression of NFκB and the ratio of Bcl-2/Bax expression were declined in the NaHS ＋cisplatin group compared with cisplatin group.[Conclusion] Hydrogen sulfide can inhibit the growth of U2OS cells at the concentration above 1.0 mmol/L and enhance the inhibition of cisplatin on proliferation of U2OS cells at the concentration above 0.8 mmol/L concentration by decreasing the transportation of NFκB to the nucleus and then increase the apoptosis of U2OS cells.

关 键 词：硫化氢 顺铂 骨肉瘤U2OS细胞 抑制 NFΚB 

分 类 号：R738.1[医药卫生—肿瘤]