鉴定调控L-plastin在非激素依赖型前列腺癌中表达的转录因子SP-1  

Identification of transcription factor SP-1 upregulating the expression of L-plastin in hormone-independent prostate cancer

作　　者：林天歆[1] 黄健[1] 尹心宝[1] 许可慰[1] 叶枫[2] 李泗耀[1] 黄海[1] 江春[1] 

机构地区：[1]中山大学附属第二医院泌尿外科 [2]中山大学附属第二医院体检中心,广东广州510120

出　　处：《中国病理生理杂志》2006年第7期1330-1334,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30500507);广东省自然科学基金资助项目(No.021846;No.05001758)

摘　　要：目的:鉴定调控L-plastin在非激素依赖型前列腺癌中表达的转录因子,进一步阐明非激素依赖型前列腺癌的发病机理。方法:在鉴定L-plastin启动子近3’端-216到1序列片段存在明显的转录活性的基础上,利用TFSEARCH软件分析该片段的序列,寻找调控L-plastin表达的非甾体激素类转录因子,并用凝胶滞后实验和超滞后实验证实结合于该片段的转录因子,利用PCR定点突变法构建删除该转录因子结合位点的重组子,并检测切除该片段序列后荧光素酶活性变化。结果:TFSEARCH软件分析表明在距转录起始点-54--41bp处含有SP-1转录因子结合位点GGTGGGGCGGGGA,凝胶滞后实验和超滞后实验表明SP-1是结合于L-plastin启动子3’端该序列的转录因子,删除SP-1结合位点后荧光素酶活性下降。结论:SP-1是促进L-plastin在非激素依赖型前列腺癌中表达的重要调控因子。AIM： To identify the non - steroid transcription factors upregulating the expression of L - plastin in hormone - independent prostate cancer, and partly elucidate the mechanism of hormone- refractory prostate cancer. METHODS： TF SEARCH software was used to analysis the possible binding sites of transcription factors in the 3＇ end of L - plastin promoter that had been identified as important part of regulation response elements. Gel shift assay and supershift assay were used to confirm the transcription factors binding the speculated response elements. PCR site - mutagenesis technique was performed to delete the binding site of transcription factor and luciferase activity assay was carried out after deletion of the binding site. RESULTS： SP - 1 respond element A located at -54 - -41 of L - plastin promoter was identified with the TF SEARCH software. Gel shift assay and supershift assay confirmed that SP - 1 was the transcription factor binding to GGTGC^GCGGGGA. Mutant deleted the SP - 1 binding - site had low - luciferase activity than that of the naive. CONCLUSION： SP - 1 plays an important role in. the up ＂regulation of L - plastin expression in hormone - independent prostate cancer.

关 键 词：前列腺肿瘤 L—plasfin 启动子 转录因子 Sp1 

分 类 号：R363[医药卫生—病理学]