Department of Pediatrics, Nanjing Maternal and Child Health Hospital of Nanjing Medical University, No. 123 Tianfei Road, Nanjing, P.R. China.
Int J Mol Med. 2010 Dec;26(6):803-11. doi: 10.3892/ijmm_00000528.
We previously identified the six-transmembrane epithelial antigen of prostate (STEAP) 4 as a novel plasma membrane protein that is up-regulated in obese patients and may play a significant role in the development of human obesity. In this study, a STEAP4-specific antibody was used to characterize the biological functions of the STEAP4 protein in human adipocytes. Cell viability assays (Trypan Blue exclusion), CCK-8 assays and cell cycle analysis showed that the STEAP4 antibody inhibited pre-adipocyte proliferation. Morphological observations by electron microscopy and confocal laser microscopy, annexin V-FITC labeling, caspase-3 and caspase-8 activity assays as well as data from quantitative real-time RT-PCR (qPCR) further determined that the STEAP4 antibody could promote apoptosis in pre-adipocytes. Based on quantitative Oil Red O staining and the expression profiles of specific markers, we demonstrated that the STEAP4 antibody did not affect adipogenesis, but the 2-deoxy-d-[3H]-glucose uptake tests showed that it induced the insulin-stimulated glucose uptake in mature human adipocytes. In conclusion, our results demonstrated that the STEAP4 antibody does not influence human adipocyte differentiation, but it is likely that the STEAP4 protein regulates proliferation and apoptosis and plays an important role in modulating the insulin sensitivity of human adipocytes.
我们之前鉴定了六跨膜上皮抗原的前列腺(STEAP)4 作为一种新型的质膜蛋白,上调在肥胖患者中,可能在人类肥胖的发展中起重要作用。在这项研究中,STEAP4 特异性抗体被用来描述 STEAP4 蛋白在人类脂肪细胞中的生物学功能。细胞活力测定(台盼蓝排除),CCK-8 检测和细胞周期分析表明,STEAP4 抗体抑制前脂肪细胞增殖。形态学观察通过电子显微镜和共聚焦激光显微镜, Annexin V-FITC 标记,caspase-3 和 caspase-8 活性测定以及定量实时 RT-PCR(qPCR)的数据进一步确定,STEAP4 抗体可以促进前脂肪细胞凋亡。基于定量油红 O 染色和特定标记物的表达谱,我们表明,STEAP4 抗体不影响脂肪生成,但 2-脱氧-d-[3H]-葡萄糖摄取试验表明,它诱导成熟的人脂肪细胞胰岛素刺激的葡萄糖摄取。总之,我们的研究结果表明,STEAP4 抗体不影响人脂肪细胞分化,但它可能是 STEAP4 蛋白调节增殖和凋亡,并在调节人脂肪细胞的胰岛素敏感性中发挥重要作用。