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TRAIL基因与顺铂协同诱导横纹肌肉瘤细胞凋亡的实验研究 被引量:1

Experimental study on synergistic effect of TRAIL gene combined with cisplatin in rhabdomyosarcoma cells
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摘要 目的探讨肿瘤坏死因子相关凋亡诱导配体基因(TRAIL)和顺铂联合应用对人RD胚胎型横纹肌肉瘤细胞的生长抑制和凋亡的诱导作用。方法将重组腺病毒载体(Ad)介导的TRAIL基因作用于人RD胚胎型横纹肌肉瘤细胞,通过电镜、MTT比色法、RT-PCR及流式细胞仪方法,观察和分析其对RD胚胎型横纹肌肉瘤细胞作用的效果,并与联合应用顺铂的效果进行对比,分析其作用机制。结果MTT法显示顺铂浓度为1.0μg/ml、5.0μg/ml和10.0μg/ml对RD细胞生长的抑制率分别为9.8%、23.4%和43.8%;按MOI值1000转染腺病毒Ad/GT-TRAIL对RD细胞生长的抑制率为41.2%,再联合Ad/PGK-GV16后,生长抑制率上升到52.5%;Ad/GT-TRAIL联合Ad/PGK-GV16再加用顺铂后的生长抑制率为72.7%。顺铂作用3d后的凋亡率为7.41%;Ad/GT-TRAIL联合Ad/PGK-GV16作用3d后的凋亡率为12.95%;Ad/GT-TRAIL联合Ad/PGK-GV16再加用顺铂作用3d后的凋亡率为23.24%,而1640对照组的凋亡率为2.19%。顺铂作用后,细胞的cFLIPmRNA表达下调,与细胞凋亡增加的改变相一致。结论Ad/GT-TRAIL能有效诱导横纹肌肉瘤细胞的凋亡,从而抑制横纹肌肉瘤细胞的生长,联合应用顺铂后能显著提高疗效。 Objective To study the synergistic induction of apoptosis by the combination of TNF-related apoptosis-inducing ligand (TRAIL) gene and cisplatin in rhabdomyosarcoma cell line. Methods Human rhabdomyosarcoma cells were transfected with adenovirus-mediated TRAIL gene (Ad/GT-TRAIL). The morphological changes, cell growth and apoptosis were measured by electron microscope, MTT method and flow cytometry respectively. The expression of cFLIP mRNA was shown by RT-PCR. Results Obvious apoptosis morphological changes in rhabdomyosarcoma cells were induced by Ad/GT-TRAIL and Ad/PGK-GVl6. The suppression percentage of cisplatin on rhabdomyosarcoma cell at 1.0 μg/ml, 5.0 μg/ml and 10.0 μg/ml was 9.8%, 23.4% and 43.8% respectively, however, the percentage of Ad/GT-TRAIL was 41.2%, and the percentage of Ad/GT-TRAIL combined with Ad/PGK-GVl6 and cispltin increased to 72.7%. The apoptotic rate of cispltin at 5.0 μg/ml was 7.41%, the rate of Ad/GT-TRAIL combined with Ad/PGK-GVl6 was 12.9%, and the rate of combination of cispltin, Ad/GT-TRAIL and Ad/PGK-GVl6 was 23.24%. The expression of cFLIP was down regulated with cisplatin, which is paralleled by the apoptotic rates. Moreover, the expression of cFLIP decreased more significantly when the cells were cultured with cispltin and TRAIL. The obvious apoptosis morphological changes, which were shrinking cellular morphology, chromatin condensation, cytoplasmic vacuoles in rhabdomyosarcoma cells, were shown by electron microscope when the cells were co-cultured with cispltin, Ad/GT-TRAIL and Ad/PGK-GVl6. Conclusion Ad/GT-TRAIL is able to induce apoptosis of rhabdomyosarcoma cells and inhibit its growth. TRAIL gene and cisplatin have synergistic effect on rhabdomyosarcoma cells. This is a potential and valuable proposal for clinical treatment of rhabdomyosacoma.
出处 《中华骨科杂志》 CAS CSCD 北大核心 2004年第7期422-426,共5页 Chinese Journal of Orthopaedics
关键词 横纹肌肉瘤 肿瘤坏死因子 基因抑制 肿瘤 Rhabdomyosarcoma Tumor necrosis factor Genes, suppressor, tumor
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参考文献10

  • 1McAllister RM;Melnyk J;Finkelstein JZ.Cultivation in vitro of cells derived from a human rhabdomyosarcoma[J],1969.
  • 2何超,胡晓彤,徐向明,方炳良.肿瘤坏死因子相关凋亡诱导配体基因对人大肠癌细胞株HT29作用的研究[J].中华肿瘤杂志,2002,24(2):133-136. 被引量:8
  • 3Zhang XD;Franco A;Myers K.Relation of TNF-related apoptosis-inducing ligand(TRAIL)receptor and FLICE-inhibitory protein expression to TRAIL-induced apoptosis of melanoma,1999.
  • 4魏金星,武玉东,刘会凡,马腾骧,赵玉千,吴长利.Fas配体基因在肾细胞癌中的表达及其意义[J].中华实验外科杂志,2000,17(3):215-216. 被引量:8
  • 5Kagawa S;Pearson SA;Ji L.A binary adenoviral vector system for expressing high levels of the proapoptotic gene bax[J],2000(1).
  • 6Irmler M;Thome M;Hahne M.Inhibition of death receptor signals by cellular FLIP[J],1997(6638).
  • 7Petak I;Douglas L;Tillman DM.Pediatric rhabdomyosarcoma cell lines are resistant to Fas-induced apoptosis and highly sensitive to TRAIL-induced apoptosis[J],2000.
  • 8Tomek S;Koestler W;Horak P.Trail-induced apoptosis and interaction with cytotoxic agents in soft tissue sarcoma cell lines[J],2003.
  • 9Bin L;Li X;Xu LG.The short splice form of Casper/c-FLIP is a major cellular inhibitor of TRAIL-induced apoptosis[J],2002.
  • 10Song JH;Song DK;Herlyn M.Cisplatin down-regulation of cellular Fas-associated death domain-like interleukin-1 beta-converting enzyme-like inhibitory proteins to restore tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in human melanoma cells[J],2003.

二级参考文献11

  • 1Lau H T,Science,1996年,273卷,109页
  • 2Kagawa S,Pearson SA,Ji L,et al.A binary adenoviral vector system for expressing high levels of the proapoptotic gene bax[].Gene Therapy.2000
  • 3Arai H,Gordon D,Nabel EG,et al.Gene transfer of Fas ligand induces tumor regression in vivo[].Proceedings of the National Academy of Sciences of the United States of America.1997
  • 4Ashkenazi A,Pai RC,Fong S,et al.Safety and antitumor activity of recombinant soluble Apo2 ligand[].The Journal of Clinical Investigation.1999
  • 5Larregina AT,Morelli AE,Dewey RA,et al.FasL induces Fas/Apo1mediated apoptosis in human embryonic kidney 293 cells routinely used to generate E1-deleted adenoviral vectors[].Gene Therapy.1998
  • 6Sheridan JP,Marsters SA,Pitti RM,et al.Control of TRAIL-induced apoptosis by a family of singaling and decoy receptors[].Science.1997
  • 7Griffith TS,Lynch DH.A molecule with multiple receptors and control mechanisms[].Current Opinion in Immunology.1998
  • 8Shinohara N,Ohashi M,Yoshida Y,et al.Construction, propagation,and titer estimation of recombinant adenoviruses carrying proapoptotic genes[].Human Gene Therapy.1998
  • 9Pan G,O′Rourke K,Chinnaiyan AM,et al.The receptor for the cytotoxic ligand TRAIL[].Science.1997
  • 10Okuyama T,Fujino M,Li XK,et al.Efficient Fas-ligand gene expression in rodent liver after intravenous injection of a recombinant adenovirus by the use of a Cre-mediated switching system[].Gene Therapy.1998

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  • 1Wang S, El-Deiry WS. TRAIL and apoptosis induction by TNF-family death receptors. Oncogene, 2003, 22: 8628-8633.
  • 2Mizutani Y, Nakanishi I-I, Yoshida O, et al. Potentiation of the sensitivity of renal cell carcinoma cells to TRAIL-mediated apoptosis by subtoxic concentrations of 5-fluorouracil. Eur J Cancer, 2002, 38:167-176.
  • 3Cuello M, Ettenberg SA, Nau MM, et al. Synergistic induction of apoptosis by the combination of trail and chemotherapy in chemoresistant ovarian cancer cells. Gynecol Oncol, 2001, 81: 380-390.
  • 4Zhang XD, Franco A, Myers K, et al. Relation of TNF-related apoptosis-inducing ligand (TRAIL) receptor and FLICE-inhibitory protein expression to TRAIL-induced apoptosis of melanoma. Cancer Res, 1999, 59: 2747-2753.
  • 5Weinmann M, Marini P, Jendrossek V, et al. Influence of hypoxia on TRAIL-induced apoptosis in tumor cells. Int J Radiat Oncol Biol Phys, 2004, 58: 386-396.
  • 6Grutter MG. Caspases: key players in programmed cell death. Curr Opin Struct Biol, 2000, 10: 649-655.
  • 7Wajant H, Pfizenmaier K, Scheurich P. TNF-related apoptosis inducing ligand (TRAIL) and its receptors in tumor surveillance and cancer therapy. Apoptosis, 2002, 7: 449-459.
  • 8Lacour S, Hammann A, Wotawa A, et al. Anticancer agents sensitize tumor cells to tumor necrosis factor-related apoptosis-inducing ligand-mediated caspase-8 activation and apoptosis. Cancer Res,2001, 61: 1645-1651.
  • 9Thomas WD, Zhang XD, Franco AV, et al. TNF-related apoptosis-inducing ligand induced apoptosis of melanoma is associated with changes in mitochondrial membrane potential and perinuclear clustering of mitochondria. J Immunol, 2000, 165: 5612-5620.
  • 10Bin L, Li X, Xu LG, et al. The short splice form of Casper/c-FLIP is a major eellular inhibitor of TRAIL-indueed apoptosis. FEBS Lett,2002, 510: 37-40.

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