CD95基因转导食管癌细胞的体外抑瘤效应

Anti-tumor effects of CD95 gene in oesophagus cancer cells transducted with CD95 gene

目的观察 CD95基因表达株表达的 CD95蛋白对体外培养食管癌细胞的抑制作用。方法 Western blot 检测 CD95基因转导前后CD95蛋白的表达水平。直接记数法描述转导株的细胞生长曲线;MTT 显色法比较转导前后细胞的体外增殖能力;药敏实验观察转导细胞对VCR、5-FU 等化疗药物的敏感性;软琼脂集落形成实验观察基因转导前后细胞的克隆形成力。结果杂交结果表明,转导株 CD95蛋白水平的表达明显高于非转导株。转导细胞的细胞倍增时间(2d)、对数生长期(3d～7d)等均体现了比非转导株(0.8d,2d～8d)更为缓慢和处于抑制状态,体外生长速度的增长指数下降,集落形成能力低下(0.9%),而对 VCR、5-FU 等化疗药物的敏感性明显增加。结论 CD95基因在食管癌细胞中处于低表达状态;通过真核表达载体的介导,CD95基因导入食管癌细胞后,能有效地表达 CD95蛋白。CD95基因转导株的表达蛋白能有效地抑制体外培养的食管癌细胞的生长。

AIM To probe into the tumor inhititory effects in oesophagus cancer cell of CD95 gene trans- duced.METHODS Finally the expressing level of CD95 gene was determined by the Western blot.The tu- mor repression function of the transfected strain were observed through cells growth curve and plating effi- ciency test.MTT assay was used to detect durg sensitivity of gene transduced cells.RESULTS The bloting results suggested,that the expressing level of protein in gene transduced cell strain was much higher than that in gene not transduced cell strain.Forthe transferred strain population doubling time was 2 days and log phase of growth was 3～7 days,plating efficiency and repression rate were 0.9% and 45% respectively. Results of MTT assay showed increasing sensitivity of gene transduced cells to CDDP,VCR,5-FU.CON- CLUSION The expressing level of CD95 gene was very low in oesophagus cancer cells.The oesophagus cancer cells transduced with reconstructed plasmid of pBK-CD95 cDNA could express CD95 mRNA and CD95 protein efficienctly.CD95 gene transgenic expression clearly repress the proliferation of oesophagus cancer cells in vitro.