姜黄素对骨肉瘤细胞增殖、凋亡影响及作用机制研究
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摘要
目的:探讨姜黄素对骨肉瘤细胞增殖、凋亡的影响及作用机制。方法:选用骨肉瘤MG-63细胞株,分别采用不同浓度的姜黄素进行处理。采用MTT比色法检测骨肉瘤MG-63细胞增殖抑制率,采用流式细胞仪检测骨肉瘤MG-63细胞凋亡情况,采用Western Blot法检测骨肉瘤MG-63细胞凋亡相关蛋白表达水平。结果:①干预12、24、48 h后,与对照组比较,姜黄素各浓度组骨肉瘤MG-63细胞增殖抑制率较高,且姜黄素5μmol/L组<10μmol/L组<15μmol/L组及20μmol/L组,差异有统计学意义(P<0.05)。②干预48 h后,与对照组比较,姜黄素各浓度组骨肉瘤MG-63细胞凋亡率较高,且姜黄素5μmol/L组<10μmol/L组<15μmol/L组,差异有统计学意义(P<0.05)。③与对照组比较,黄素各浓度组骨肉瘤MG-63细胞P53、B细胞淋巴瘤/白血病-2(Bcl-2)、Caspase-9前体(proCaspase-9)蛋白表达水平较低,且姜黄素5μmol/L组>10μmol/L组>15μmol/L组,差异有统计学意义(P<0.05)。④与对照组比较,姜黄素各浓度组骨肉瘤MG-63细胞Bcl相关蛋白(Bax)、细胞色素C(Cyt-C)、天冬氨酸特异性半胱氨酸蛋白酶-3(Caspase-3)蛋白表达水平较高,且姜黄素5μmol/L组<10μmol/L组<15μmol/L组,差异有统计学意义(P<0.05)。结论:姜黄素能剂量依赖性的抑制骨肉瘤MG-63细胞增殖,并促进其凋亡,其作用可能与下调P53、Bcl-2、proCaspase-9蛋白表达,上调Bax、Cyt-C、Caspase-3蛋白表达,激活线粒体凋亡途径有关。
Objectives:To analysis the effects and mechanism of anticancer drug curcumin on proliferation and apoptosis of osteosarcoma cells. Methods:Osteosarcoma MG-63 cells were selected and treated with different concentrations of curcumin. The osteosarcoma MG-63 cells proliferation inhibition rate was detected by MTT colorimetric method,apoptosis of osteosarcoma MG-63 cells was detected by flow cytometry,the expression levels of apoptosis related protein were detected by Western Blot method in osteosarcoma MG-63 cells. Results:(1) After the intervention of 12 h,24 h and 48 h,compared with control group,the proliferation inhibition rate of osteosarcoma MG-63 cells were higher in each concentration of curcumin group,and curcumin 5 μmol/L group<10 μmol/L group<15 μmol/L group and 20 μmol/L group,the differences were statistically significant(P<0.05).(2)After the intervention of 48 h,compared with control group,the apoptosis rate of osteosarcoma MG-63 cells were higher in each concentration of curcumin group,and curcumin 5 μmol/L group<10 μmol/L group<15 μmol/L group,the differences were statistically significant(P<0.05).(3)Compared with control group,the expression of P53,B cell lymphoma/leukemia-2(Bcl-2),Caspase-9 precursor(proCaspase-9)protein in osteosarcoma MG-63 cells were lower in each concentration of curcumin group,and curcumin 5 μmol/L group>10 μmol/L group>15 μmol/L group,the differences were statistically significant(P<0.05).(4)Compared with control group,the expression of Bcl related protein(Bax),cytochrome C(Cyt-C),aspartate specific cysteine protease-3(Caspase-3)protein in osteosarcoma MG-63 cells were higher in each concentration of curcumin group,and curcumin 5 μmol/L group<10 μmol/L group<15 μmol/L group,the differences were statistically significant(P<0.05). Conclusion:Anticancer drug curcumin can inhibit the proliferation and promote apoptosis of osteosarcoma MG-63 cells in a dose-dependent manner,the effects may be related to down-regulation of P53,Bcl-2,proCaspase-9 protein expression,up-regulation of Bax,Cyt-C,Caspase-3 protein expression,and the activation of mitochondrial apoptosis pathway.
Objectives:To analysis the effects and mechanism of anticancer drug curcumin on proliferation and apoptosis of osteosarcoma cells. Methods:Osteosarcoma MG-63 cells were selected and treated with different concentrations of curcumin. The osteosarcoma MG-63 cells proliferation inhibition rate was detected by MTT colorimetric method,apoptosis of osteosarcoma MG-63 cells was detected by flow cytometry,the expression levels of apoptosis related protein were detected by Western Blot method in osteosarcoma MG-63 cells. Results:(1) After the intervention of 12 h,24 h and 48 h,compared with control group,the proliferation inhibition rate of osteosarcoma MG-63 cells were higher in each concentration of curcumin group,and curcumin 5 μmol/L group<10 μmol/L group<15 μmol/L group and 20 μmol/L group,the differences were statistically significant(P<0.05).(2)After the intervention of 48 h,compared with control group,the apoptosis rate of osteosarcoma MG-63 cells were higher in each concentration of curcumin group,and curcumin 5 μmol/L group<10 μmol/L group<15 μmol/L group,the differences were statistically significant(P<0.05).(3)Compared with control group,the expression of P53,B cell lymphoma/leukemia-2(Bcl-2),Caspase-9 precursor(proCaspase-9)protein in osteosarcoma MG-63 cells were lower in each concentration of curcumin group,and curcumin 5 μmol/L group>10 μmol/L group>15 μmol/L group,the differences were statistically significant(P<0.05).(4)Compared with control group,the expression of Bcl related protein(Bax),cytochrome C(Cyt-C),aspartate specific cysteine protease-3(Caspase-3)protein in osteosarcoma MG-63 cells were higher in each concentration of curcumin group,and curcumin 5 μmol/L group<10 μmol/L group<15 μmol/L group,the differences were statistically significant(P<0.05). Conclusion:Anticancer drug curcumin can inhibit the proliferation and promote apoptosis of osteosarcoma MG-63 cells in a dose-dependent manner,the effects may be related to down-regulation of P53,Bcl-2,proCaspase-9 protein expression,up-regulation of Bax,Cyt-C,Caspase-3 protein expression,and the activation of mitochondrial apoptosis pathway.
引文
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[12]Marquardt J U,Gomezquiroz L,Arreguin Camacho L O,et al. Curcumin effectively inhibits oncogenic NF-kB signaling and restrains stemness features in liver cancer[J]. Journal of Hepatology,2015,63(3):661-669.
[13]Xu X,Chen D,Ye B,et al. Curcumin induces the apoptosis of non-small cell lung cancer cells through a calcium signaling pathway[J]. International Journal of Molecular Medicine,2015,35(6):1610-1616.
[14]原向伟,黄秀芳.姜黄素对骨肉瘤细胞增殖和凋亡的影响[J].中国医药科学,2017,7(13):30-32.
[15]Pei-Pei Li,Wei He,Ping-Fan Yuan,et al. Celastrol Induces Mitochondria-Mediated Apoptosis in Hepatocellular Carcinoma Bel-7402 Cells[J]. American Journal of Chinese Medicine,2015,43(1):137-148.
[2]吴发帅,高枫,浦飞飞,等.长链非编码RNA在骨肉瘤中的研究进展[J].华中科技大学学报(医学版),2017,61(6):719-723.
[3]郭征.我国骨肉瘤治疗的现状与问题及发展方向[J].中国骨与关节杂志,2015,4(5):338-342.
[4]耿磊,陈继营,许猛,等.骨肉瘤的治疗进展[J].中国矫形外科杂志,2015,23(21):1975-1978.
[5]唐华科,曾星,胡志全.姜黄素抗癌机制的研究进展[J].现代泌尿生殖肿瘤杂志,2016,41(3):121-122.
[6]李学春,许江燕,陶茂灿,等.姜黄素对人骨肉瘤细胞的生长抑制及对MAPK基因的调控作用研究[J].中华中医药学刊,2017,36(2):428-431.
[7]Yang J L. Investigation of osteosarcoma genomics and its impact on targeted therapy:an international collaboration to conquer human osteosarcoma[J].癌症(英文版),2014,33(12):575-580.
[8]张明,胡向阳,杜远立.骨肉瘤相关分子机制研究进展[J].海南医学,2017,28(11):1826-1829.
[9]刘云霞,徐叶峰,李梦芸,等.清化复肝汤防治骨肉瘤甲氨蝶呤化疗性肝损伤的临床研究[J].浙江中医杂志,2017,52(2):83-84.
[10]李伟锋,蒋建兰.姜黄素药理作用的研究现状[J].中国临床药理学杂志,2017,33(10):957-960.
[11]苗淑涵,高晋生.姜黄素药理作用的研究进展[J].光明中医,2017,32(15):2284-2287.
[12]Marquardt J U,Gomezquiroz L,Arreguin Camacho L O,et al. Curcumin effectively inhibits oncogenic NF-kB signaling and restrains stemness features in liver cancer[J]. Journal of Hepatology,2015,63(3):661-669.
[13]Xu X,Chen D,Ye B,et al. Curcumin induces the apoptosis of non-small cell lung cancer cells through a calcium signaling pathway[J]. International Journal of Molecular Medicine,2015,35(6):1610-1616.
[14]原向伟,黄秀芳.姜黄素对骨肉瘤细胞增殖和凋亡的影响[J].中国医药科学,2017,7(13):30-32.
[15]Pei-Pei Li,Wei He,Ping-Fan Yuan,et al. Celastrol Induces Mitochondria-Mediated Apoptosis in Hepatocellular Carcinoma Bel-7402 Cells[J]. American Journal of Chinese Medicine,2015,43(1):137-148.