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目的构建小鼠CD154真核表达质粒,观察其转染膀胱癌细胞后对癌细胞体内成瘤性的影响。方法逆转录聚合酶链反应(RTPCR)法扩增T739小鼠CD154cDNA,重组到pcDNA3.1/Zeo(+)构成重组质粒,经阳离子脂质体转染小鼠膀胱移行细胞癌细胞BTT739,将转染后的细胞接种T739小鼠,观察肿瘤在体内的生长情况。结果RTPCR产物进行凝胶电泳可见0.8kb处的目的基因条带,序列测定结果正确;CD154重组质粒转染BTT739细胞后能够在肿瘤细胞内表达;转染CD154的肿瘤细胞在小鼠体内不能成瘤。结论成功构建小鼠CD154真核表达质粒,将其转染肿瘤细胞后能抑制肿瘤细胞的体内成瘤性。
Objective To construct a murine CD154 eukaryotic expression plasmid and observe its effect on the tumorigenicity of cancer cells after transfection into bladder cancer cells. Methods CD154 cDNA of T739 mice was amplified by reverse transcription polymerase chain reaction (RTPCR). Recombinant plasmid pcDNA3.1 / Zeo (+) was constructed and transfected into bladder transitional cell carcinoma cell line BTT739 by cationic liposome. The stained cells were seeded in T739 mice to observe the growth of the tumor in vivo. Results The target gene of 0.8kb was obtained by gel electrophoresis. The result of sequencing was correct. The recombinant plasmid of CD154 could be expressed in tumor cells after transfected with BTT739 cells. The tumor cells transfected with CD154 could not form tumor in mice . Conclusion The mouse CD154 eukaryotic expression plasmid was successfully constructed and transfected into tumor cells can inhibit the tumorigenicity of tumor cells in vivo.