Yang Fu-Lan, Chen Fang-Zhi, Wan Xin-Xing, Zhou Xi, Zhou Mei-Juan, Chen Han-Chun, Fu Jun-Jiang, Zhang Dian-Zheng
Department of Biochemistry, School of Life Sciences and The State Key Laboratory of Medical Genetics, Central South University, Changsha, Hunan 410013, China.
Department of Urology, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, China.
Biomed Res Int. 2015;2015:325026. doi: 10.1155/2015/325026. Epub 2015 Sep 1.
Using the phage display biopanning technique, we have previously identified a heptapeptide KLWVIPQ which specifically binds to the surface of the IFN-α-sensitive but not the IFN-α-resistant CML cells. The effects of this heptapeptide on the IFN-α-sensitive CML cells were investigated in the present study. IFN-α-sensitive KT-1/A3 and IFN-α-resistant KT-1/A3R CML cells were transfected by pEGFP-KLWVIPQ expression vector and/or induced by IFN-α. WST-1 cell proliferation assay, flow cytometry, and western blotting were performed to determine the effects of this heptapeptide and/or IFN-α on CML cells. The viability of the KT-1/A3 cells was inhibited and apoptosis was induced by either expression of the heptapeptide KLWVIPQ or IFN-α treatment with concurrent upregulation of P53 and downregulation of P210(bcr/abl). However, these effects were not observed in the IFN-α-resistant KT-1/A3R cells. These results suggest that the heptapeptide KLWVIPQ shares a similar mechanism with IFN-α in the regulation of CML cell growth and apoptosis, implying that the heptapeptide KLWVIPQ could be a novel target to go further into mechanisms of IFN-α sensitivity and/or resistance in CML.
利用噬菌体展示生物淘选技术,我们之前鉴定出一种七肽KLWVIPQ,它能特异性结合干扰素-α敏感的慢性粒细胞白血病(CML)细胞表面,但不结合干扰素-α耐药的CML细胞表面。本研究调查了这种七肽对干扰素-α敏感的CML细胞的影响。用pEGFP-KLWVIPQ表达载体转染干扰素-α敏感的KT-1/A3和干扰素-α耐药的KT-1/A3R CML细胞,和/或用干扰素-α诱导。进行WST-1细胞增殖试验、流式细胞术和蛋白质印迹法,以确定这种七肽和/或干扰素-α对CML细胞的影响。七肽KLWVIPQ的表达或干扰素-α处理均可抑制KT-1/A3细胞的活力并诱导其凋亡,同时伴随P53上调和P210(bcr/abl)下调。然而,在干扰素-α耐药的KT-1/A3R细胞中未观察到这些效应。这些结果表明,七肽KLWVIPQ在调节CML细胞生长和凋亡方面与干扰素-α具有相似的机制,这意味着七肽KLWVIPQ可能是进一步深入研究CML中干扰素-α敏感性和/或耐药性机制的新靶点。