期刊文献+

非增殖型和靶向增殖型腺病毒携带TRAIL基因治疗肝癌的实验研究

GENE THERAPY OF HEPATOCELLULAR CARCINOMA USING REPLICATION-DEFECTIVE AND TARGETED TUMOR-SPECIFIC REPLICATION-COMPETENT ADENOVIRUS CARRYING HUMAN TRAIL GENE IN VITRO
下载PDF
导出
摘要 目的比较携带人TRAIL基因的增殖型腺病毒(CNHK500-hTRAIL)和非增殖型腺病毒(Ad-hTRAIL)对TRAIL基因的表达以及对SMMC-7721肝癌细胞的杀伤能力。方法通过病毒增殖实验评估增殖型腺病毒CNHK500-hTRAIL的选择性增殖能力。通过MTT实验,评估增殖型腺病毒CNHK500-hTRAIL以及非增殖型腺病毒Ad-hTRAIL对人正常肝细胞株L02、人肝癌细胞株SMMC-7721的杀伤能力。采用ELISA法检测CNHK500-hTRAIL和Ad-hTRAIL感染SMMC-7721肝癌细胞后TRAIL基因的表达情况。以及通过流式细胞术(FCM)检测其对细胞早期凋亡的影响。结果CNHK500-hTRAIL能选择性地在SMMC-7721细胞内大量增殖,感染96 h后增殖达225137倍,在极低的MO I值(MO I=0.1)即可大量杀伤SMMC-7721细胞,明显强于Ad-hTRAIL,而对L02细胞无明显杀伤。CNHK500-hTRAIL和Ad-hTRAIL感染SMMC-7721细胞后,其TRAIL基因表达量,前者是后者的近10倍;CNHK500-hTRAIL可选择性地诱导SMMC-7721细胞早期凋亡,其能力显著高于Ad-hTRAIL。结论靶向增殖型腺病毒载体携带TRAIL基因对肿瘤细胞的杀伤能力和目的基因的表达,明显优于传统的非增殖型腺病毒载体,应用前景广阔。 Objective To evaluate the therapeutic effect and the expression level of a tumor-specific replication-competent adenovirus and a replication-defective adenovirus expression human soluble TRAIL(hTRAIL) gene on hepatocellular carcinoma cell line SMMC-7721 in vitro. Methods Virus replication assay was performed to evaluate the selective replication ability of CNHK500- hTRAIL. The cytotoxicity of replication- competent adenovirus CNHK500-hTRAIL and replication-defective adenovirus Ad-hTRAIL were evaluated by MTT in HCC cell line SMMC-7721 and human normal hepatocyte line L02. ELISA assay were used to determine the expression level of TRAIL. Flow Cytometry ( FCM ) was used to detect the early apoptosis induced by CNHK500-hTRAIL and Ad- hTRAIL. Results CNHK500-hTRAIL could selectively proliferated in SMMC-7721 with an increase of 225137-fold in 96h post-infection. CNHK500-hTRAIL could kill SMMC-7721 cells at a very low MOI( MOI = 0. 1 ), and could induce SMMC-7721 cell lines obviousapoptosis, while it had no significant effect on L02. The expressing level of TRAIL in CNHK500-hTRAIL treated SMMC-7721 cell lines was much higher than that in Ad-hTRAIL treated one (almost 10-fold). The ability of therapeutic effect, transgene expression and apoptosis inducing were stronger than that of replication-defective Ad-hTRAIL. Conclusion Targeted replication-competent adenovirus carrying human TRAIL gene is more effective than that of replication-defective adenovirus carrying human TRAIL gene in both cytotoxicity and efficiency of gene transfer in HCC, and holds great promise in the area of cancer therapy.
出处 《肝胆外科杂志》 2009年第2期136-140,共5页 Journal of Hepatobiliary Surgery
基金 南京军区重大专项课题基金资助(No.07Z006)
关键词 增殖型腺病毒 TRAIL 基因治疗 凋亡 肝细胞癌 Replication-competent adenovirus TRAIL Gene-therapy Apoptosis Hepatocellular carcinoma
  • 相关文献

参考文献1

二级参考文献10

  • 1Kim D, Martuza RL, Zwiebel J. Replication-selective virotherapy for cancer: Biological principles, risk management and future directions. Nature Med. 2001; 7:781.
  • 2Xin Yuan Liu. A new anticancer strategy-genetic and virological treatnent of cancer. Chinese J Cancer Biother 2001; 8: 1.
  • 3Zhao L, Gu J, Dong A, et al. Potent antitumor activity of oncolytic adenovirus expressing mda-7/IL-24 for colorectal cancer. Hum Gene Ther 2005; 16: 845-58.
  • 4Griffith TS, Chin WA, Jackson GC, et al. Intracellular regulation of TRAIL-induced apoptosis inhuman melanoma cells. J.Immunol. 1998: 161: 2833-40.
  • 5Pitti RM, Marsters SA, Ruppert S, et al. Induction of apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family. J Biol Chem 1996; 271: 12687-90.
  • 6Liu XY, Qiu SB, Zou WG, et al. Effective gene-virotherapy for complete rradication of tumor mediated by the combination of hTRAIL (TNFSF 10) and plasminogen k5. Mol Ther 2005; 11:531-41.
  • 7Pei Z, Chu L, Zou W, et al. An oncolytic adenoviral vector of Smac increases antitumor activity of TRAIL against HCC in human cells and in mice. Hepatology 2004; 39:1371-81.
  • 8Zou W, Luo C, Zhang Z, et al. A novel oncolytic adenovirus targeting to telomerase activity in tumor cells with potent. Oncogene 2004; 23: 457-64.
  • 9Yun-xia Tang, Yu Chen, Jin-fa Gu, et al. Cancer gene-virotherapy of Ad-TERT-TRAIL. Chinese J Cancer 2005; 24: 536-42.
  • 10Zhang Q, Nie M, Sham J, et al. Effective gene-viral therapy for telomerase-positive cancers by selective replicative-competent adenovirus combining with endostatin gene. Cancer Res 2004;64: 5390-97.

共引文献15

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部