论文部分内容阅读
构建了以癌胚抗原（ＣＥＡ）基因启动子控制的ＨＳＶ－ＴＫ和ＥＣＣＤ的表达质粒ＰＣＥＡ－ＴＫ和ｐＣＥＡ－ＣＤ．将它们分别与ｐＳＶ２－ｎｅｏ共转染人结肠癌细胞株ＬｏＶｏ和人宫颈癌细胞株ＨｅＬａ，Ｇ４１８筛选得到细胞克隆ＬｏＶｏ／ＣＥＡ－ＴＫ、ｔｏＶｏ／ＣＥＡ－ＣＤ、ＨｅＬａ／ＣＥＡ－ＴＫ和ＨｅＬａ／ＣＥＡ－ＣＤ．与野生型ＬｏＶｏ细胞相比，ＬｏＶｏ／ＣＥＡ－ＴＫ和ＬｏＶｏ／ＣＥＡ－ＣＤ形态无明显改变，生长曲线也相似，但对ＧＣＶ或５－ＦＣ的细胞毒的敏感性分别提高了２０００倍或７００信而ＨｅＬａ／ＣＥＡ－ＴＫ（或ＨｅＬａ／ＣＥＡ－ＣＤ）仍对低浓度ＧＣＶ（或５－ＦＣ）不敏感．以上结果显示了应用组织专一性表达的自杀基因治疗人结肠癌的可能性．
The expression plasmids PCEA-TK and pCEA-CD of HSV-TK and ECCD controlled by the promoter of carcinoembryonic antigen (CEA) gene were constructed. These cells were co-transfected with pSV2-neo and transfected into human colon carcinoma cell line LoVo and human cervical carcinoma cell line HeLa. G418 cell clones were selected to obtain LoVo/CEA-TK, toVo/CEA-CD, HeLa/CEA-TK and HeLa/CEA. -CD. Compared with wild-type LoVo cells, the morphology of LoVo/CEA-TK and LoVo/CEA-CD did not change significantly and the growth curve was similar, but the sensitivity to cytotoxicity of GCV or 5-FC was increased by 2000 times or 700 letters, respectively. However, HeLa/CEA-TK (or HeLa/CEA-CD) is still insensitive to low concentrations of GCV (or 5-FC). The above results show the possibility of using tissue-specific suicide gene therapy for human colon cancer.