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目的制备前列腺非雄激素依赖启动子(PSES)介导的鸟氨酸脱羧酶(ODC)和S-腺苷甲硫氨酸脱羧酶(AdoMetDC)双反义腺病毒。方法构建pShuttle-Basic-PSES-AdoMetDC-ODC-PolyA穿梭载体,并与载体pAd-PL进行体外重组,包装成重组腺病毒。将重组腺病毒转染至前列腺癌、直肠癌等癌细胞中,噻唑兰比色(MTT)法观察其对细胞增殖的影响,Western Blot检测重组腺病毒对细胞中ODC和AdoMetDC表达的抑制作用。结果成功构建了前列腺特异的ODC和AdoMetDC双反义RNA表达载体,并包装制备出能特异的在前列腺细胞中表达产生ODC和AdoMetDC反义RNA的重组腺病毒。该重组腺病毒具有前列腺组织特异性。结论构建的前列腺非雄激素依赖启动子介导的ODC、AdoMetDC双反义腺病毒具有组织特异性。
OBJECTIVE: To prepare prostatic androgen-independent promoter (PSES) -mediated ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (AdoMetDC) double antisense adenovirus. Methods The pShuttle-Basic-PSES-AdoMetDC-ODC-PolyA shuttle vector was constructed and recombined with vector pAd-PL in vitro and packaged into recombinant adenovirus. The recombinant adenovirus was transfected into cancer cells such as prostate cancer and rectum. The effect of recombinant adenovirus on cell proliferation was observed by MTT assay. The inhibitory effect of recombinant adenovirus on ODC and AdoMetDC expression was detected by Western Blot. RESULTS: Prostate-specific ODC and AdoMetDC double antisense RNA expression vectors were successfully constructed and packaged to produce recombinant adenovirus specific for expression of ODC and AdoMetDC antisense RNA in prostate cells. The recombinant adenovirus has prostate tissue specificity. Conclusions The constructed non-androgen-dependent prostatic adenocarcinoma of the ODC and AdoMetDC double antisense adenovirus has tissue specificity.