放射诱导的肿瘤特异性嵌合启动子研究  

Study of cancer-specific chimeric promoters induced by irradiation

作　　者：熊杰[1] 周云峰[1] 孙文洁[1] 王伟峰[1] 廖正凯[1] 周福祥[1] 谢丛华[1] 

机构地区：[1]武汉大学中南医院放化疗科肿瘤生物学行为湖北省重点实验室湖北省肿瘤医学临床研究中心,430071

出　　处：《中华放射医学与防护杂志》2010年第2期152-155,共4页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金（30672438）;湖北省创新群体项目（2006ABC009）

摘　　要：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4～6Gy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2Gy与单次6Gy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。Objective To combine the radio-inducible CArG element with cancer-specific human telomerase reverse transcriptase（hTERT） gcne 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 ceils （ HeLa, A549, and MHCC97 cells） and nomal cells（hEL cells） were detected by using lucifcrase-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 gent expression was observed after 3 fractionated doses of 2 Gy compared with a single close 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 genc therapy.

关 键 词：人端粒酶反转录酶启动子 CArG元件 基因治疗 

分 类 号：R73-3[医药卫生—肿瘤]