| 田梅,金光辉,朴春姬,等.辐射诱导表达载体pEgr-hPTEN的构建及其体外抗肿瘤作用的研究[J].中华放射医学与防护杂志,2003,23(6):423-426.TIAN Mei,JIN Guanghui,PIAO Chunji,et al.Study on construction of pEgr-hPTEN expression vector induced by irradiation and its anti-tumor effect in vitro[J].Chin J Radiol Med Prot,2003,23(6):423-426 |
| 辐射诱导表达载体pEgr-hPTEN的构建及其体外抗肿瘤作用的研究 |
| Study on construction of pEgr-hPTEN expression vector induced by irradiation and its anti-tumor effect in vitro |
| 投稿时间：2003-06-27 |
| DOI： |
| 中文关键词: 人抑癌基因hPTEN pEgr-hPTEN重组表达载体 基因-放射治疗 |
| 英文关键词:hPTEN pEgr-hPTEN expression vector Gene -radio therapy |
| 基金项目:国家自然科学基金资助项目(30170290) |
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| 中文摘要: |
| 目的 克隆人野生型抑癌基因PTEN/MMAC1cDNA序列、构建辐射诱导表达载体pEgr-hPTEN并研究其体外稳定转染联合X-射线照射对恶性胶质瘤细胞SHG44增殖的抑制作用。方法 利用R-TnestedPCR法从正常人胎盘组织中扩增出约1200bp的PTEN片段,并将其重组入pcDNA3.1Egr载体中,构建为表达质粒pEgr-hPTEN,体外转染肿瘤细胞并经G418筛选稳定转染细胞株SHG44sPTEN,接受不同剂量X射线照射,观察基因放射联合作用对肿瘤细胞生长的影响。结果 经全自动测序证明克隆得到了野生型PTEN基因;辐射诱导表达载体pEgr-hPTEN构建正确;稳定转染联合X-射线照射可明显抑制肿瘤细胞的恶性增殖,5Gy以内随吸收剂量的增加,肿瘤抑制作用更明显。结论 本研究成功克隆了人野生型抑癌基因PTEN/MMAC1cDNA序列并构建了辐射诱导表达载体pEgr-hPTEN,体外基因放射联合治疗具有明显的肿瘤抑制作用。本研究为提高临床恶性肿瘤放疗疗效奠定了理论和实验基础。 |
| 英文摘要: |
| Objective To clone the cDNA of human tumor suppressor gene-PTEN ,construct pEgr-hPTEN e xpression vector induced by irradiation and study its inhibitory effect on proli feration of malignant glioma cell line SHG-44 transfected steadily with pEgr-h PTEN after different doses of X-ray irradiation. Methods A DNA fragment about 1 200 bp ,PTEN, was amplified from human placenta tissues by using RT-nes ted PCR and was cloned into pUCm-T vector after automatic sequencing, the n the fragment was inserted into a vector pcDNA3 1-Egr to construct an express i on vector pEgr-hPTEN. pEgr-hPTEN was transfected into SHG-44 cells in vitro . Stably transfected cell line SHG-44-sPTEN was selected through G418. The inhi bitor effect on SHG-44-sPTEN was observed after different doses of X-ray irra diation in vitro. Results The PTEN cDNA has been cloned correctly and its expre ssion vector pEgr-hPTEN was also constructed. Growth of SHG-44 cells was inhi bited significantly by stable pEgr-hPTEN transfection combined with X-ray irra diation. With the increase of dose , the inhibitory effect was enhanced within 5 Gy. Conclusion Human tumor suppressor gene-PTEN cDNA has been cloned and its e xpression vector has been constructed. The tumor was inhibited significantly by gene-radiotherapy in vitro. The result provides the theoretical and experim ental basis for improvement of clinical radiotherapeutic effect on tumors . |
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