Wu Chen, Shan Yaojun, Liu Xinxia, Song Wenqian, Wang Jiali, Zou Minji, Wang Min, Xu Donggang
College of Life Sciences, Hebei University, Baoding, Hebei, 071002, P R China.
J Biosci. 2009 Sep;34(3):389-95. doi: 10.1007/s12038-009-0045-z.
Insulin-like growth factor binding protein-3 (IGFBP-3) is known to induce apoptosis in an insulin-like growth factor (IGF)-dependent and IGF-independent manner, but the mechanism underlying the IGF-independent effects remains unclear. Polypeptide N-acetylgalactosaminyltransferase 14 (GalNAc-T14) is a novel IGFBP-3 binding partner. In this paper, small interference RNA (siRNA) targeting GalNAc-T14 was used to examine whether GalNAc-T14 affects the apoptotic action of IGFBP-3. Using semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) and western blot analysis, we determined that GalNAc-T14 expression was downregulated by the siRNA directed against GalNAc-T14. Apoptosis analysis of IGFBP-3-overexpressing cells treated with siRNA against GalNAc-T14 was performed to determine if GalNAc-T14 was specifically involved in IGFBP-3 signalling. The results, as determined by flow cytometric analysis and caspase-3 assay, showed that the extent of apoptosis induced by IGFBP-increased with RNA interference (RNAi) knockdown of GalNAc-T14. Our data suggest that GalNAc-T14 influences the apoptotic action of IGFBP-3 and might mediate the signalling pathway of IGFBP-3. Experiments to determine the role of GalNAc-T14 in the regulation of apoptosis induced by IGFBP-3 are under way.
胰岛素样生长因子结合蛋白-3(IGFBP-3)已知可通过胰岛素样生长因子(IGF)依赖性和IGF非依赖性方式诱导细胞凋亡,但其IGF非依赖性作用的潜在机制仍不清楚。多肽N-乙酰半乳糖胺基转移酶14(GalNAc-T14)是一种新型的IGFBP-3结合伴侣。在本文中,使用靶向GalNAc-T14的小干扰RNA(siRNA)来研究GalNAc-T14是否影响IGFBP-3的凋亡作用。通过半定量逆转录聚合酶链反应(RT-PCR)和蛋白质印迹分析,我们确定针对GalNAc-T14的siRNA下调了GalNAc-T14的表达。对用针对GalNAc-T14的siRNA处理的IGFBP-3过表达细胞进行凋亡分析,以确定GalNAc-T14是否特异性参与IGFBP-3信号传导。通过流式细胞术分析和caspase-3检测确定的结果表明,IGFBP诱导的细胞凋亡程度随着GalNAc-T14的RNA干扰(RNAi)敲低而增加。我们的数据表明,GalNAc-T14影响IGFBP-3的凋亡作用,并可能介导IGFBP-3的信号通路。确定GalNAc-T14在IGFBP-3诱导的细胞凋亡调节中的作用的实验正在进行中。