前列腺癌HRE.CArG.HSV-TK重组腺病毒载体基因放射治疗的体外研究  

In vitro research of recombinant adenovirus vector HRE.CArG.HSV-TK gene and radiation in prostate cancer cells

作　　者：魏玲[1] 宋现让[1] 孙菊杰[1] 王兴武[1] 宋宝[1] 郑燕[1] 

机构地区：[1]山东省肿瘤医院病理科(山东省放射肿瘤学重点实验室),山东济南250117

出　　处：《中国现代医学杂志》2010年第21期3245-3248,3253,共5页China Journal of Modern Medicine

基　　金：山东省医学科学院科研基金项目(No:2006-14)

摘　　要：目的观察HRE.CArG.HSV-TK重组腺病毒载体Ad.HRE.CArG.HSV-TK联合放射对体外培养的前列腺癌细胞LNCaP和PC-3生长和凋亡的影响。方法 50 MOI重组腺病毒体外转导细胞,Real–timeRT-PCR检测TK基因mRNA表达水平,MTT法检测腺病毒转导联合放射对常氧(37℃,19%氧气,5%二氧化碳)和乏氧(37℃,0.5%氧气,5%二氧化碳)状态下细胞的生长抑制作用,流式细胞术检测凋亡(AnnexinV-FITC染色)。结果 50MOI腺病毒体外转导细胞48 h后,TK mRNA表达较对照组明显增加(P<0.05),乏氧和/或放射处理后TK mRNA表达增加更显著。腺病毒体外转导和/或联合射线照射后,细胞抑制率显著增加并可介导细胞凋亡(P<0.05)。乏氧处理时上述效应更明显。结论腺病毒载体联合放射可显著抑制体外人前列腺癌细胞生长并诱导凋亡,为前列腺癌进一步的基因-放射治疗研究奠定了基础。【Objective】 To observe the influence of HRE.CArG.HSV-TK recombinant adenovirus vector-Ad.HRE.CArG.HSV-TK combined with radiation on the growth and apoptosis of prostate cancer cells in vitro.【Methods】 Cells were transducted with 50 MOI adenovirus vector.TK mRNA expression was detected by Real-time RT-PCR.The growth inhibition effect of adenovirus transduction combined with radiation on cells under normoxia（37℃,19% O2,5%CO2） and hypoxia（37℃,0.5% O2,5% CO2） was detected by MTT assay.Apoptosis（Annexin V-FITC staining）was determined by flow cytometry.【Results】 After LNCaP and PC-3 cells were transducted by 50MOI recombinant adenovirus for 48 h,TK mRNA expression increased dramatically compared with control（P 0.05）.These effects became more obvious after hypoxia and or radiation treatment.The inhibition rate was upregulated and apoptosis was induced in cells transducted with adenovirus and/or combined with radiation（P 0.05）.Above effects were more obvious undergoing hypoxia treatment.【Conclusion】 Adenovirus vector combined with radiation can significantly suppress the growth and induce apoptosis in prostate cancer cells.This provides the foundation for further gene radiation therapy of prostate cancer.

关 键 词：前列腺癌 腺病毒 HSV-TK基因 乏氧 放射 

分 类 号：R737.25[医药卫生—肿瘤]