Yu Guiping, Shen Zhenya, Chen Guoqiang, Teng Xiaomei, Hu Yanqiu, Huang Bin
Department of Cardiovascular Surgery, The First Affiliated Hospital of Soochow University, No. 188, Shi Zi Rd., Soochow, 215006, People's Republic of China.
Tumour Biol. 2013 Feb;34(1):91-8. doi: 10.1007/s13277-012-0514-0. Epub 2012 Sep 15.
The purposes of this study were to investigate the effects of phosphatidylethanolamine-binding protein 4 (PEBP4) on the cell growth, proliferation, apoptosis, and invasion of non-small cell lung cancer (NSCLC) cells and to provide evidence for future treatment options for NSCLC. Western blot assays were performed to examine PEBP4 protein expression levels in NSCLC cell lines (HCC827, A549, NCI-H661, NCI-H292, and 95-D) and a normal human bronchial epithelial (HBE) cell line. A PEBP4 shRNA expression vector was constructed and transfected into HCC827 cells. Subsequently, the effects of PEBP4 on the cell viability, cell cycle distribution, apoptosis levels, and invasion properties of HCC827 cells were analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, flow cytometry analyses, and transwell invasion assays. In addition, the effects of PEBP4 on the expression of proteins including cyclin D1, p53, Bcl-2, MMP-2, and MMP-9 were investigated. PEBP4 was highly expressed in lung cancer cells (HCC827, A549, NCI-H661, NCI-H292, and 95-D), but its expression was low in HBE cells. Cell viability, cell proliferation, and invasion of HCC827 cells in the PEBP4 knockdown group were significantly lower than that in the negative control and blank control groups (p < 0.05), and there were no significant differences between the negative and blank control groups in terms of cell viability, cell proliferation, apoptosis, and invasion. In HCC827 cells, the expression levels of cyclin D1, Bcl-2, MMP-2, and MMP-9 in the PEBP4 knockdown group were significantly lower (p < 0.05), and the expression of p53 protein was significantly higher than that in the negative and blank control groups (p < 0.05). There were no significant differences between the negative and blank control groups in the expression levels of cyclin D1, p53, Bcl-2, MMP-2, and MMP-9. In conclusion, PEBP4 enhanced HCC827 cell proliferation and invasion ability and inhibited apoptosis. Decreased PEBP4 expression may play a role in the reduced invasion ability and increased apoptosis of the human NSCLC cell line HCC827.
本研究的目的是探讨磷脂酰乙醇胺结合蛋白4(PEBP4)对非小细胞肺癌(NSCLC)细胞的生长、增殖、凋亡和侵袭的影响,并为NSCLC的未来治疗方案提供依据。采用蛋白质免疫印迹法检测NSCLC细胞系(HCC827、A549、NCI-H661、NCI-H292和95-D)及正常人支气管上皮(HBE)细胞系中PEBP4蛋白的表达水平。构建PEBP4短发夹RNA(shRNA)表达载体并转染至HCC827细胞。随后,采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法、流式细胞术分析和Transwell侵袭实验分析PEBP4对HCC827细胞活力、细胞周期分布、凋亡水平和侵袭特性的影响。此外,还研究了PEBP4对细胞周期蛋白D1、p53、Bcl-2、基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)等蛋白表达的影响。PEBP4在肺癌细胞(HCC827、A549、NCI-H661、NCI-H292和95-D)中高表达,但在HBE细胞中低表达。PEBP4基因敲低组HCC827细胞的活力、增殖和侵袭能力均显著低于阴性对照组和空白对照组(p < 0.05),阴性对照组和空白对照组在细胞活力、增殖、凋亡和侵袭方面无显著差异。在HCC827细胞中,PEBP4基因敲低组细胞周期蛋白D1、Bcl-2、MMP-2和MMP-9的表达水平显著降低(p < 0.05),p53蛋白的表达显著高于阴性对照组和空白对照组(p < 0.05)。阴性对照组和空白对照组在细胞周期蛋白D1、p53、Bcl-2、MMP-2和MMP-9的表达水平上无显著差异。综上所述,PEBP4增强了HCC827细胞的增殖和侵袭能力并抑制了凋亡。PEBP4表达降低可能在人NSCLC细胞系HCC827侵袭能力降低和凋亡增加中发挥作用。