中华放射医学与防护杂志
中華放射醫學與防護雜誌
중화방사의학여방호잡지
Chinese Journal of Radiological Medicine and Protection
2010年
2期
152-155
,共4页
熊杰%周云峰%孙文洁%王伟峰%廖正凯%周福祥%谢丛华
熊傑%週雲峰%孫文潔%王偉峰%廖正凱%週福祥%謝叢華
웅걸%주운봉%손문길%왕위봉%료정개%주복상%사총화
人端粒酶反转录酶启动子%CArG元件%基因治疗
人耑粒酶反轉錄酶啟動子%CArG元件%基因治療
인단립매반전록매계동자%CArG원건%기인치료
Human telomerase reverse transcriptase gene promoter%CArG elements%Gene therapy
目的 将放射敏感性的CArG元件与肿瘤特异性的人端粒酶反转录酶(hTERT)基因启动子相结合,构建新的嵌合启动子.方法 构建含不同CArG元件数量的嵌合hTERT启动子,在放射线诱导下(单剂量或分割照射),利用荧光素酶报告基因检测其在肿瘤细胞(HeLa、A549、MHCC97细胞)及正常细胞(hEL细胞)中的活性.结果 包含6个CArG元件的hTERT嵌合启动子较含其他数量CArG的嵌合启动子显示了更好的放射诱导性,并在4～6 Gy时达到顶峰,而构建的嵌合启动子在正常的hEL细胞中活性很低.在肿瘤细胞中,3次2 Gy与单次6 Gy剂量照射相比较,报告基因的表达相当.结论 包含6个CArG元件的肿瘤特异性嵌合启动子具备良好的放射诱导性,这种嵌合的启动子系统具有肿瘤基因治疗的潜力.
目的 將放射敏感性的CArG元件與腫瘤特異性的人耑粒酶反轉錄酶(hTERT)基因啟動子相結閤,構建新的嵌閤啟動子.方法 構建含不同CArG元件數量的嵌閤hTERT啟動子,在放射線誘導下(單劑量或分割照射),利用熒光素酶報告基因檢測其在腫瘤細胞(HeLa、A549、MHCC97細胞)及正常細胞(hEL細胞)中的活性.結果 包含6箇CArG元件的hTERT嵌閤啟動子較含其他數量CArG的嵌閤啟動子顯示瞭更好的放射誘導性,併在4～6 Gy時達到頂峰,而構建的嵌閤啟動子在正常的hEL細胞中活性很低.在腫瘤細胞中,3次2 Gy與單次6 Gy劑量照射相比較,報告基因的錶達相噹.結論 包含6箇CArG元件的腫瘤特異性嵌閤啟動子具備良好的放射誘導性,這種嵌閤的啟動子繫統具有腫瘤基因治療的潛力.
목적 장방사민감성적CArG원건여종류특이성적인단립매반전록매(hTERT)기인계동자상결합,구건신적감합계동자.방법 구건함불동CArG원건수량적감합hTERT계동자,재방사선유도하(단제량혹분할조사),이용형광소매보고기인검측기재종류세포(HeLa、A549、MHCC97세포)급정상세포(hEL세포)중적활성.결과 포함6개CArG원건적hTERT감합계동자교함기타수량CArG적감합계동자현시료경호적방사유도성,병재4～6 Gy시체도정봉,이구건적감합계동자재정상적hEL세포중활성흔저.재종류세포중,3차2 Gy여단차6 Gy제량조사상비교,보고기인적표체상당.결론 포함6개CArG원건적종류특이성감합계동자구비량호적방사유도성,저충감합적계동자계통구유종류기인치료적잠력.
Objective To combine the radio-inducible CArG element with cancer-specific human telomerase reverse transcriptase(hTERT) gene promoter,and to construct the novel chimeric promoters.Methods The synthetic hTERT promoters containing different number of radio-inducible CArG elements were constructed.and the activities of the promoters in the cancer cells (HeLa,A549,and MHCC97 cells) and nomal cells(hEL cells) were detected by using luciferase-reporter assays afer the treatment of irradiation(a single or fractionated irradiation dose).Results Synthetic promoter containing 6 repeated CArG units was better in radio-inducibility than any other promoters containing different number of CArG units.and nearly maximum levels obtained at 4-6 Gy.The very low activities of the chimeric promoters could be detected in normal hEL cells.A similar level of reporter gene expression was observed after 3 fractionated doses of 2 Gy compared with a single dose of 6 Gy in cancer cells.Conclusions The cancerspecific chimeric promoter containing 6 CArG elements showes the best radio-response,and the chimeric promoter system has the potential in cancer gene therapy.
