Yang Shaoxing, Qi Fei, Tang Chuanhao, Wang Hong, Qin Haifeng, Li Xiaoyan, Li Jianjie, Wang Weixia, Zhao Changyun, Gao Hongjun
Department of Pulmonary Oncology, Affiliated Hospital, Academy of Military Medical Science, Beijing 100071, P.R. China.
Department of Respiratory Medicine, Chinese PLA General Hospital, Beijing 100853, P.R. China.
Oncol Lett. 2017 Feb;13(2):898-904. doi: 10.3892/ol.2016.5502. Epub 2016 Dec 14.
Cluster of differentiation (CD) 147 is a transmembrane glycoprotein that is highly expressed at the tumor cell surface, which stimulates fibroblasts to produce a large number of matrix metalloproteinases and promotes tumor invasion and metastasis and tumor-induced angiogenesis. The present study investigated the functions and the role of CD147 in the human lung carcinoma A549 cell line. The present study constructed expression and interference [small interfering (si) RNA] lentiviral vectors of CD147, which established stable overexpression and low expression of CD147 in the A549 cell line, named A549-CD147 and A549-siCD147, respectively. The differences in biological features between various levels of CD147 expression in A549 cells was investigated by cell counting kit-8 (CCK-8), Transwell, scratch and lumen formation assays. The results of the CCK-8 assay revealed that A549-CD147 cell proliferation was significantly increased and A549-siCD147 cell proliferation was decreased compared with the control groups. The A549-CD147 cells had the largest number of cells penetrating the Matrigel in the Transwell assay, which indicates that upregulation of CD147 expression increases the infiltration capacity of cells. The scratch assay revealed that A549-CD147 cells have the highest capacity for migration, while A549-siCD147 cells have the lowest. Quantitative polymerase chain reaction and western blot analysis demonstrated that vascular endothelial growth factor (VEGF) expression was proportional to the expression level of CD147 at the mRNA and protein level. The lumen formation assay revealed that the number of vessel lumens that human umbilical vein endothelial cells formed in the A549-CD147 cell supernatant was increased compared with the A549-siCD147 cells. Collectively, the present results suggest that CD147 is important in the promotion of lung carcinoma cell proliferation, invasion and metastasis and the upregulation of VEGF, which stimulates the angiogenesis of lung carcinoma. In conclusion, CD147 may be a potential target in the treatment of lung carcinoma.
分化簇(CD)147是一种跨膜糖蛋白,在肿瘤细胞表面高度表达,可刺激成纤维细胞产生大量基质金属蛋白酶,促进肿瘤侵袭、转移及肿瘤诱导的血管生成。本研究探讨了CD147在人肺癌A549细胞系中的功能及作用。本研究构建了CD147的表达和干扰[小干扰(si)RNA]慢病毒载体,分别在A549细胞系中建立了CD147的稳定过表达和低表达,命名为A549-CD147和A549-siCD147。通过细胞计数试剂盒-8(CCK-8)、Transwell、划痕和管腔形成实验,研究了A549细胞中不同水平CD147表达的生物学特性差异。CCK-8实验结果显示,与对照组相比,A549-CD147细胞增殖显著增加,A549-siCD147细胞增殖减少。Transwell实验中,A549-CD147细胞穿透基质胶的细胞数量最多,这表明CD147表达上调可增加细胞的浸润能力。划痕实验显示,A549-CD147细胞迁移能力最强,而A549-siCD147细胞迁移能力最弱。定量聚合酶链反应和蛋白质印迹分析表明,血管内皮生长因子(VEGF)在mRNA和蛋白质水平的表达与CD147的表达水平呈正相关。管腔形成实验显示,与A549-siCD147细胞相比,人脐静脉内皮细胞在A549-CD147细胞上清液中形成的血管管腔数量增加。总体而言,目前的结果表明,CD147在促进肺癌细胞增殖、侵袭和转移以及上调VEGF从而刺激肺癌血管生成方面具有重要作用。总之,CD147可能是肺癌治疗的潜在靶点。