松乳菇多糖对人骨肉瘤MG-63细胞Caspase-3、Caspase-9、Bax和Bcl-2蛋白表达的影响
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摘要
目的研究松乳菇多糖体外对人骨肉瘤MG-63细胞生长、凋亡及凋亡相关蛋白的影响。方法体外培养MG-63细胞,将不同浓度的松乳菇多糖作用于细胞,MTT法检测不同时间点和不同浓度的松乳菇多糖对细胞生长的抑制作用; Annexin V-FITC/PI双染流式细胞仪检测对细胞凋亡的影响,并分析细胞周期; Western blot法检测Caspase-3、Caspase-9、Bax和Bcl-2蛋白的表达。结果松乳菇多糖可时间和浓度依赖性地抑制MG-63细胞的生长;通过阻滞G2-M期促进MG-63细胞凋亡;促进Caspase-3、Caspase-9和Bax蛋白的表达,抑制Bcl-2蛋白的表达。结论松乳菇多糖体外可抑制人骨肉瘤MG-63细胞的生长,诱导其凋亡,其诱导凋亡的作用机制与调节Caspase-3,Caspase-9,Bax和Bcl-2蛋白的表达有关。
Objective To investigate the activity of polysaccharide fromLactarius deliciosus Gray on the proliferation,apoptosis and apoptosis-related protein in human osteosarcoma MG-63 cells.Methods MG-63 cells were cultured with polysaccharide fromLactarius deliciosus Gray in vitro.MTT was used to measure cell growth inhibition.Annexin V-FITC/PI was used to determine the apoptosis rate and the cell cycle.Western blot was used to detect the expression of Caspase-3,Caspase-9,Bax and Bcl-2.Results Polysaccharide fromLactarius deliciosus Gray inhibited the proliferation of MG-63 cells in the concentrationand time-dependent manner.Moreover,polysaccharide elevated the apoptosis rates by blocked in G2-M phase,promoted the expression of Caspase-3,Caspase-9 and Bax and inhibited the expression of Bcl-2.Conclusion Polysaccharide fromLactarius deliciosus Gray can inhibit the growth of MG-63 cells and induce its apoptosis,which may be related to modulation of Caspase-3,Caspase-9,Bax and Bcl-2 expression.
Objective To investigate the activity of polysaccharide fromLactarius deliciosus Gray on the proliferation,apoptosis and apoptosis-related protein in human osteosarcoma MG-63 cells.Methods MG-63 cells were cultured with polysaccharide fromLactarius deliciosus Gray in vitro.MTT was used to measure cell growth inhibition.Annexin V-FITC/PI was used to determine the apoptosis rate and the cell cycle.Western blot was used to detect the expression of Caspase-3,Caspase-9,Bax and Bcl-2.Results Polysaccharide fromLactarius deliciosus Gray inhibited the proliferation of MG-63 cells in the concentrationand time-dependent manner.Moreover,polysaccharide elevated the apoptosis rates by blocked in G2-M phase,promoted the expression of Caspase-3,Caspase-9 and Bax and inhibited the expression of Bcl-2.Conclusion Polysaccharide fromLactarius deliciosus Gray can inhibit the growth of MG-63 cells and induce its apoptosis,which may be related to modulation of Caspase-3,Caspase-9,Bax and Bcl-2 expression.
引文
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[14]李冷,钟晓琴,卢传坚,等.紫草素对人角质形成细胞HaCaT增殖抑制以及细胞毒性的初步研究[J].时珍国医国药,2017,28(4):857-860.
[2]MIRABELLO L,PFEIFFER R,MURPHY G,et al.Height at diagnosis and birth-weight as risk factors for osteosarcoma[J].Cancer Causes Con,2011,22(6):899-908.
[3]SANCHEZ-BARCELO E J,MEDIAVILLA M D,TAN D X,et al.Scientific basis for the potential use of melatonin in bone diseases:osteoporosis and adolescent idiopathic scolisis[J].JOsteoporos,2010,2010:830-831.
[4]钱叶,丁祥,曾益春,等.松乳菇多糖刺激免疫细胞增殖及诱导肿瘤细胞凋亡的研究[J].食品科学,2017,38(5):220-226.
[5]卯晓岚.中国大型真菌[M].郑州:河南科学技术出版社,2000:369.
[6]敖常伟,惠明,李忠海,等.松乳菇营养成分分析及松乳菇多糖的提取分离[J].2003,24(9):77-79.
[7]邱美珍,傅胜才,周望平,等.松乳菇菌丝多糖对小鼠免疫调节作用研究[J].黑龙江畜牧兽医科技版,2010,21(3):137-139.
[8]刘露,宋波,赵大群,等.松乳菇多糖(LDG-A)对S180荷瘤小鼠生理生化指标的影响[J].西华师范大学学报(自然科学版),2016,37(4):380-384.
[9]SONAA E,USHA S,JAIN J.An ex vivo study of selenium,genistein on the morphological and nuclear changes in anticancer drug-induced apoptosis in human peripheral blood lymphocytes[J].Bio Factors,2013,39(3):279-293.
[10]何雅军,舒建昌,吕霞,等.Annexin V-FITC/PI流式细胞术检测姜黄素诱导的肝星状细胞凋亡[J].实用医学杂志,2009,25(1):28-30.
[11]陈巧云,王业秋,曲岩,等.桃叶珊瑚苷对光损伤HaCaT细胞Bax,Bcl-2,Caspase-3表达的影响[J].现代生物医学进展,2017,17(1):24-27.
[12]CHO H M,SUN W.Control of mitochondrial dynamics by Fasinduced caspase-8 activation in hippocampal neurons[J].Exp Neurobiol,2015,24(3):219-225.
[13]刘亚莉,易佳丽,刘春英.补中益气汤含药血清干预A549/DDP细胞Bad、NF-κb、Caspase-9表达的实验研究[J].时针国医国药,2017,28(7):1548-1550.
[14]李冷,钟晓琴,卢传坚,等.紫草素对人角质形成细胞HaCaT增殖抑制以及细胞毒性的初步研究[J].时珍国医国药,2017,28(4):857-860.