【摘要】 目的 :了解分泌肽序列在HepG2细胞中对人内皮抑素基因 (hEndostatin)的cDNA表达和分泌差异的影响。方法 :构建不含hIL - 2分泌肽序列的真核表达载体pBlast-hEndo质粒 ,转染人的HepG2细胞株 ,用RT -PCR的方法检测HepG2 (pBlast-hIL2 -hEndo) ,HepG2 (pBlast-hEndo) ,HepG2 (pBlast-Mcs)和HepG2中hEndostatin的mR NA表达水平 ,及制备 4种细胞的总蛋白和收集各自的培养上清 ,进行Westernblot分析蛋白表达和分泌的差异。结果 :发现HepG2 (pBlast-hIL2 -hEndo)中hEndostatin的mRNA的水平显著高于HepG2 (pBlast -hEndo)。而仅在HepG2(pBlast-hIL - 2 -hEndo)的细胞总蛋白和上清 ,及HepG2 (pBlast -hEndo)的细胞总蛋白中检测到hEndostatin表达 ,在其它各株细胞总蛋白及上清中 ,未检测到hEndostatin。结论 :hIL - 2分泌肽序列能促进hEndostatin基因在HepG2细胞中表达及分泌。更多还原

【Abstract】 AIM: The role of human interleukin-2(IL-2) signal peptide sequence in the effect of human Endostatin (hEndostatin) expression and secretion was investigated in HeG2 cells. METHODS: RT-PCR and Western-blotting were conduct to observe mRNA level difference of hEndostatin gene, its protein expression and secretion level difference between with hIL-2 signal peptide sequence and without it. RESULTS: mRNA level of hEndostatin gene in HepG2 (pBlast-hIL2-hEndo) cells was higher than that in HepG2(pBlast-hEndo)( P<0.05 ). hEndostatin protein was detected only in HepG2 (pBlast-hIL2-hEndo) cells and its medium, and not in other HepG2 cells and medium. CONCLUSION: Human interleukin-2(IL-2) signal peptide sequence facilitate to increase mRNA level of hEndostatin gene, its protein expression and secretion in HepG2 cells.更多还原