藤黄酸对骨肉瘤中端粒酶逆转录酶和P53蛋白的影响
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摘要
目的研究藤黄酸对骨肉瘤(HOS)中端粒酶逆转录酶(hTERT)和P53蛋白的影响,为藤黄酸治疗HOS的临床应用提供理论依据。方法以0.125、0.25、0.5、1、2、4、8μmol/L剂量的藤黄酸分别作用于对数生长期的HOS细胞,用CCK-8法测定细胞存活率,通过免疫组化SP法测定hTERT、P53蛋白的表达情况,应用生物医学影像分析软件Image Pro Plus和Image J对免疫组化图片进行半定量分析。结果随着藤黄酸作用时间的延长和浓度的增高,HOS细胞存活率降低;随着藤黄酸作用时间的延长,hTERT的表达减弱,P53蛋白的表达明显增强。结论在藤黄酸作用下,藤黄酸剂量越高,作用时间越长,HOS细胞存活率越低;HOS细胞存活率越低,hTERT表达越少,P53蛋白表达越多;hTERT与P53蛋白两者属非同步关系。
Objective To investigate the effects of gambogic acid(GA) on human telomerase reverse transcriptase(hTERT) and P53 protein in human osteosarcoma(HOS) cells,and to provide theoretical evidences for the clinical application of GA in osteosarcoma treatment.Methods Log phase HOS cells were influenced using GA of 0.125,0.25,0.5,1,2,4,and 8 μmol/L,the cell survival was measured using CCK-8 method,while hTERT and P53 protein expression was measured by immunohistochemistry.Immunohistochemical images was also semi-quantitative analyzed using Image Pro Plus and Image J biomedical image analysis software.Results The HOS cell survival was reduced with GA in time-and concentration-dependent manners.Meanwhile,hTERT was down-regulated while P53 was up-regulated by GA in time-dependent manner.Conclusion HOS cell survival is negatively correlated to GA dose and action time.P53 protein expression is enhanced by GA,while hTERT is weakened.
Objective To investigate the effects of gambogic acid(GA) on human telomerase reverse transcriptase(hTERT) and P53 protein in human osteosarcoma(HOS) cells,and to provide theoretical evidences for the clinical application of GA in osteosarcoma treatment.Methods Log phase HOS cells were influenced using GA of 0.125,0.25,0.5,1,2,4,and 8 μmol/L,the cell survival was measured using CCK-8 method,while hTERT and P53 protein expression was measured by immunohistochemistry.Immunohistochemical images was also semi-quantitative analyzed using Image Pro Plus and Image J biomedical image analysis software.Results The HOS cell survival was reduced with GA in time-and concentration-dependent manners.Meanwhile,hTERT was down-regulated while P53 was up-regulated by GA in time-dependent manner.Conclusion HOS cell survival is negatively correlated to GA dose and action time.P53 protein expression is enhanced by GA,while hTERT is weakened.
引文
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[15]GU H,WANG X,RAO S,et al.Gambogic acid mediates apopto-sis as a p53 inducer through down-regulation of mdm2 in wild-type p53-expressing cancer cells[J].Mol Cancer Ther,2008,7(10):3298-3305.
[2]ARINAGA M,SHIMIM S,GOTOH K,et al.Expression of hu-man telomerase subunit genes in primary lung cancer and its clini-cal significance[J].Ann Thorac Stag,2000,70(2):401-406.
[3]KIDO A,TSUJIUNCHI T,MORISHITA T,et al.Telomerase ac-tivity correlates with growth of transplantable osteosarcomas in ratstreated with cis-diammine dichloroplatinum or the angiogenesisinhibitor AGM-1470[J].Jpn J Cancer Res,1998,89(10):1074-1081.
[4]RAMAKRISHNAN S,EPPENBERGER U,MUELLER H,et al.Expression profile of the putative catalgtic subunit of the telomerasegene[J].Cancer Res,1998,58(4):622-625.
[5]YOKOYAMA Y,TAKAHASHI Y,SHINOHARA A,et al.The5'-end of hTERT mRNA is a good target for hammerhead ri-bozyme to suppress telomerase activity[J].Biochem Biophys ResCommun,2000,273(1):316-321.
[6]SHARMA G G,GUPTA A,WANG H,et al.hTERT associateswith human telomeres and enhances genomic stability and DNA re-pair[J].Oncogene,2003,22(1):131-146.
[7]MASUTOMI K,YU E Y,KHURTS S,et al.Telomerase main-tains telomere structure in normal human cells[J].Cell,2003,114(2):241-245.
[8]朱悦,郑剑玲,张斌,等.端粒酶催化亚单位和cmyc在骨肉瘤中的表达[J].中国肿瘤临床,2003,30(1):23-25.
[9]何云燕,夏云.p53水平对端粒酶逆转录酶表达的调节作用[J].第三军医大学学报,2006,28(4):345-347.
[10]ZHU Y,SHU X,CHEN J,et al.Effect of Helicobacter pylori e-radication on oncogenes and cell proliferation[J].Eur J Clin In-vest,2008,38(9):628-633.
[11]BROWN T,ALDOUS W,LANCE R,et al.The association be-tween telomerase,p53 and clinical staging in colorectal cancer[J].Am J Surg,1998,175(5):364-366.
[12]AKESHIMA K,KIGAWA T,TAKAHASHINI M,et al.Telomer-ase activity and p53-dependent apoptosis in ovarian cancer cells[J].Br J Cancer,2001,84(11):1551-555.
[13]ZHAO Q,YANG Y,YU J,et al.Posttranscriptional regulation ofthe telomerase hTERT by gambogic acid in human gastric carcino-ma 823 cells[J].Cancer Lett,2008,262(2):223-231.
[14]XU X,LIU Y,WANG L,et al.Gambogic acid induces apoptosisby regulating the expression of Bax and Bcl-2 and enhancingcaspase-3 activity in human malignant melanoma A375 cells[J].Int J Dermatol,2009,48(2):186-192.
[15]GU H,WANG X,RAO S,et al.Gambogic acid mediates apopto-sis as a p53 inducer through down-regulation of mdm2 in wild-type p53-expressing cancer cells[J].Mol Cancer Ther,2008,7(10):3298-3305.