BMS-345541对人骨肉瘤细胞MG63凋亡及其作用机制
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摘要
目的研究BMS-345541对人骨肉瘤细胞MG63凋亡的影响及其可能的机制。方法以0、2、4、6、8、10μmol/L BMS-345541处理MG63细胞24、48 h后,用CCK-8法检测细胞存活率。用0、2、4、8μmol/L药物作用细胞24 h后,以PI染色法观察细胞凋亡情况、流式细胞仪检测细胞凋亡。以同样浓度作用细胞48 h,Western-blot检测cleaved Caspase-3蛋白表达情况。结果 (4-10)μmol/L的BMS-345541均可抑制MG63细胞的增殖,其抑制效应呈剂量-时间依赖性,48 h最低存活率只有(0.412±0.024)(P<0.05)。PI染色荧光显微镜观察结果发现:随药物浓度增高,相同视野下正常细胞数逐渐降低,凋亡细胞数逐渐增高。流式细胞仪分析:0、2、4、8μmol/L的作用BMS-345541 24 h后,凋亡率分别为(5.2±0.78)%、(5.82±0.82)%、(8.86±1.71)%、(11.01±2.69)%,与对照组相比,8μmol/L组差异有统计学意义(P<0.05)。Western blot结果显示MG 63细胞中的cleaved caspase-3蛋白表达量上调,与对照组相比,2、4、8μmol/L组差异均有统计学意义(P<0.05)。结论 BMS-345541可抑制MG63细胞增殖,并诱导其发生凋亡,其作用机制可能与cleaved caspase-3表达上调有关。
Objective To study the effect of BMS-345541 on apoptosis of human osteosarcoma cell MG63 and its possible mechanism. Methods Treated MG63 cells with 0(blank control),2,4,6,8,10 μmol/L BMS-345541 for 24 h and 48 h,respctively,cell viability were detected by CCK-8 method.The group of 0,2,4,8 μmol/L BMS-345541 effect on cells for 24 h,respectively. Apoptosis was respected by PI staining,apoptosis rate of cell was analyzed by flow cytometry.Treated cells with the same concentration for 48 h,cleaved caspase-3 protein was identified by Western-blot. Results(4-10)μmol/L of BMS-345541 inhibited the proliferation of MG63 cells in a dose-dependent and time-dependent manner. The minimum survival rate was only(0.412±0.024)(P<0.05). PI staining showed that with the increase of drug concentration,the number of normal cells decreased gradually and the number of apoptotic cells increased gradually. Flow cytometry analysis show:The percentages of apoptpsis after the treatment with BMS-345541 at 0,2,4,8 μmol/L for 24 h were(5.2±0.78)%,(5.82±0.82)%,(8.86±1.71)% and(11.01±2.69)%,respectively. The data compared with the control group,the group of8 μmol/L was statistically significant(P<0.05). Western blot results showed that the expression of cleaved caspase-3 protein in MG 63 cells was up-regulated. The data compared with the control group,the concentration of 2,4,8 μmol/L was statistically significant(P<0.05). Conclusion BMS-345541 can inhibit the proliferation of MG63 cells and induce apoptosis.The mechanism may be related to the up-regulation of cleaved caspase-3 expression
Objective To study the effect of BMS-345541 on apoptosis of human osteosarcoma cell MG63 and its possible mechanism. Methods Treated MG63 cells with 0(blank control),2,4,6,8,10 μmol/L BMS-345541 for 24 h and 48 h,respctively,cell viability were detected by CCK-8 method.The group of 0,2,4,8 μmol/L BMS-345541 effect on cells for 24 h,respectively. Apoptosis was respected by PI staining,apoptosis rate of cell was analyzed by flow cytometry.Treated cells with the same concentration for 48 h,cleaved caspase-3 protein was identified by Western-blot. Results(4-10)μmol/L of BMS-345541 inhibited the proliferation of MG63 cells in a dose-dependent and time-dependent manner. The minimum survival rate was only(0.412±0.024)(P<0.05). PI staining showed that with the increase of drug concentration,the number of normal cells decreased gradually and the number of apoptotic cells increased gradually. Flow cytometry analysis show:The percentages of apoptpsis after the treatment with BMS-345541 at 0,2,4,8 μmol/L for 24 h were(5.2±0.78)%,(5.82±0.82)%,(8.86±1.71)% and(11.01±2.69)%,respectively. The data compared with the control group,the group of8 μmol/L was statistically significant(P<0.05). Western blot results showed that the expression of cleaved caspase-3 protein in MG 63 cells was up-regulated. The data compared with the control group,the concentration of 2,4,8 μmol/L was statistically significant(P<0.05). Conclusion BMS-345541 can inhibit the proliferation of MG63 cells and induce apoptosis.The mechanism may be related to the up-regulation of cleaved caspase-3 expression
引文
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[2]Mirabello L,Troisi RJ,Savage SA.Osteosarcoma incidence and survival rates from 1973 to 2004:data from the Surveillance,Epidemiology,and End Results Program[J].Cancer,2009,115(7):1531-1543.
[3]马善新,李世德.骨肉瘤综合治疗的研究进展[J].蛇志,2013(02):194-196.
[4]Jaffe N.Osteosarcoma:review of the past,impact on the future.The American experience[J].Cancer Treat Res,2009,152:239-262.
[5]Burke J R,Pattoli M A,Gregor K R,et al.BMS-345541 is a highly selective inhibitor of I kappa B kinase that binds at an allosteric site of the enzyme and blocks NF-kappa B-dependent transcription in mice[J].J Biol Chem,2003,278(3):1450-1456.
[6]田崛,陈显凌,庄英婷,等.BMS-345541对急性粒细胞白血病细胞DNA损伤修复的影响[J].中国药理学通报,2015(06):763-769.
[7]Huang YC,Pan M,Liu N,et al.Effects of nuclear factor-kappa B and ERK signaling transduction pathway inhibitors on human melanoma cell proliferation in vitro[J].Oncol Lett,2015,10(5):3233-3237.
[8]Battula VL,Nguyen K,Sun J,et al.IKK inhibition by BMS-345541 suppresses breast tumorigenesis and metastases by targeting GD2+cancer stem cells[J].Oncotarget,2017.
[9]Ping H,Yang F,Wang M,et al.IKK inhibitor suppresses epithelial-mesenchymal transition and induces cell death in prostate cancer[J].Oncol Rep,2016,36(3):1658-1664.
[10]Hideshima H,Yoshida Y,Ikeda H,et al.IKKbeta inhibitor in combination with bortezomib induces cytotoxicity in breast cancer cells[J].Int J Oncol,2014,44(4):1171-1176.
[11]Ritter J,Bielack SS.Osteosarcoma.Ann Oncol,2010,21 Suppl 7:i320-i325.
[12]汪新媛,田京.骨肉瘤化疗研究进展[J].国际骨科学杂志,2010(03):159-161.
[13]Liu JJ,Yang CL,Chen YQ,et al.Mechanism of SGC-7901 apoptosis induced by Houttuynia cordata thunb subterraneous stem extraction[J].Chinese Pharmacological Bulletin,2014,30(2):257-261.