盐酸夫拉平度对人骨肉瘤细胞MG63细胞周期及凋亡的影响  

Flavopiridol suppresses cell cycle and induces apoptosis in human osteosarcoma MG63 cell line

作　　者：李洁[1] 施佳辰[2] 李啸然[1] 李月白[1] 全碧波[1] 宋石[1] 王义生[3] 

机构地区：[1]郑州大学基础医学院生物化学教研室,郑州450001 [2]郑州大学基础医学院,郑州450001 [3]郑州大学第一附属医院骨科,郑州450052

出　　处：《郑州大学学报（医学版）》2014年第4期517-520,共4页Journal of Zhengzhou University(Medical Sciences)

摘　　要：目的：观察不同浓度盐酸夫拉平度（FP）对人骨肉瘤MG63细胞增殖活力、细胞周期及细胞凋亡的影响。方法：用含体积分数10%胎牛血清的RPMI 1640培养液常规培养MG63细胞,取对数生长期增殖旺盛细胞分为对照组和不同浓度FP干预组。MTT比色法测定不同浓度的FP对细胞增殖活性的影响;流式细胞术检测不同浓度的FP干预24和48 h后细胞周期变化和凋亡情况;RT-PCR测定不同浓度FP干预后,细胞周期相关基因表达的变化;Western blot检测细胞内CDK2蛋白表达变化。结果：50-2 000 nmol/L FP可抑制MG63细胞增殖活性,且有时间和浓度依赖性（F时间=319.470,F浓度=616.988,P均〈0.001）;不同浓度的FP分别干预MG63细胞不同时间后均可引起细胞G2期阻滞（F24 h=6.805,F48 h=12.000,P均〈0.001）,凋亡率增加（P均〈0.001）,且可明显降低MG63细胞中CDK2、Survivin mRNA及CDK2蛋白的表达（FmRNA=415.517、15.068,F蛋白=50.405,P均〈0.001）。结论：FP可抑制人骨肉瘤MG63细胞增殖及周期相关蛋白的表达,同时使细胞G2/M期阻滞并诱导细胞凋亡。Aim： To investigate the influence of flavopiridol（ FP） on proliferation,cell cycle and cell apoptosis in human osteosarcoma MG63 cell lines. Methods： The cells were cultured with RPMI1640 medium containing 10% of fetal bovine serum as usual,and treated with different concentration FP. MTT assay was used to detect the influence of different dose FP treatment on cell proliferation. Flow cytometry（ FCM） was used to determine the cycle distribution and apoptosis status of the cells which treated with various concentration FP for 24 and 48 h respectively. RT-PCR was used to test the mRNA expression of cell cycle related genes after the FP intervention. The protein expression of CDK2 in cells after FP treatment was detected by Western blot. Results： FP could inhibit the cell proliferation in the range of 50 to 2 000 nmol /L,the inhibition rate also increased with time at the range from 24 to 72 h（ Fconcentration= 616. 988,Ftime= 319. 470,P〈0. 001）. The different concentrations of FP that respectively acting on the MG63 cells for different times could arrested MG63 cells at G2 phase obviously（ F24 h= 6. 805,F48 h= 12. 000,P〈0. 001） and increased apoptosis rate（ P〈0. 001）,and could also decrease the mRNA level of CDK2 and Survivin obviously in MG63 cells and down-regulate the expression of CDK2 protein as well（ FmRNA= 415. 517,15. 068,Fprotein= 50. 405,P〈0. 001）. Conclusion： The CDK inhibitor flavopiridol can inhibit the cell proliferation and the expression of cell cycle related protein in human osteosarcoma MG63 cell,and induce the cell G2/M phase arrest and the cell apoptosis.

关 键 词：盐酸夫拉平度 MG63 凋亡 CDK 

分 类 号：R733[医药卫生—肿瘤]