轉殖病毒胸腺嘧啶激脢基因於p53圖變形人類非小細胞肺癌細胞株之研究

The study of p53-mutated human non-small cell lung cancer cells transfected with HerpesSimplex Virus type I- thymidine kinase (HSV-tk) cDNA

黃智欣(Huang Chih Hsin)

指導教授 ： 方剛

國立臺灣師範大學/理學院/生物學系/碩士(2002年)

摘要

無資料

關鍵字

非小細胞肺癌細胞 ； 賽默芬 ； 旁觀者效應 ； 老化

並列摘要

In view of the functional loss of tumor suppressors being associated to drug resistance in most of lung cancers, we have demonstrated dose-dependent ganciclovir (GCV)-induced cytotoxicity following delivery of recombinant prodrug herpes simplex virus type I thymidine kinase (HSV-tk) cDNA in highly metastatic and tumorigenic human non-small-cell-lung cancer (NSCLC) cells H1437 with mutant p53 (R267P). It is found that cells with acquisition of HSV-tk cDNA conferred GCV susceptibility to the selected clones and the effect proved proportional to the level of HSV-TK expressed both in vivo and in vitro. The inhibition of cell proliferation in culture and the regression of xenograft tumors in nude mice were demonstrated caused by triggering senescence program as a result of GCV exposure in the selected HSV-tk clones. The potent bystander effects both in vivo and in vitro systems was found correlated with HSV-TK expressed accordingly. In addition, H1437 cells underwent ecotopic transfer of HSV-tk lead to S- and G2/M-phase arrest after GCV induction prior to emergence of senescence-like phenotype that expression of the senescence marker SA-β-gal and enlarged and fattened morphology. And we also observed that transfectant cells were growth-retarded after drug treatment by PKH67 fluorescence detection. To address more in-depth mechanism leading to senescence phenotype, we identified two steps prior to cell death as triggered by senescence progression: (a) decreased expression and activity of telomerase (b) escalation of endogenous cell cycle modulators, cyclin A and cyclin B1. Taken all the work together, we concluded that HSV-tk/GCV system constitutes an effective treatment in human NSCLC cells with mutated p53 and the pathway leading to cell death through senescence activation ought to become an alternative and effective approach in restraining growth of human NSCLC cells lacking functional p53.

並列關鍵字

non-small cell lung cancer cells ； Herpes Simplex Virus type I- thymidine kinase ； ganciclovir ； bystander effect ； senescence

參考文獻

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