hIL-2分泌肽序列在HepG2细胞中对hEndostatin表达和分泌差异的影响

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mRNA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。

Signal peptide sequence of human interleukin-2 influenced hEndostatin gene expression and protein secretion in HepG2 cells

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.