背景:TetraspaninsCD151,跨膜4超家族蛋白,已被确定参与各种癌症的起始。然而,CD151在非小细胞肺癌(NSCLC)中的确切功能尚不清楚.这里,我们通过靶向EGFR/ErbB2来解决CD151在NSCLC中的促肿瘤作用,这有利于肿瘤增殖,移民和入侵。
方法:首先,用RT-PCR检测NSCLC组织和细胞系中CD151的mRNA表达水平。同时,通过蛋白质印迹法分析CD151及其相关蛋白。然后通过免疫组织化学验证NSCLC样本及其配对的邻近肺组织中CD151的表达水平。通过共免疫沉淀评估蛋白质相互作用。流式细胞术用于细胞周期分析。CCK-8,EdU公司,和克隆形成试验用于分析细胞活力。伤口愈合,Transwell迁移,和基质胶侵袭试验用于评估肿瘤细胞的运动性。探讨CD151在体内的作用,肺癌异种移植小鼠模型。
结果:在NSCLC组织和细胞系中发现高CD151表达,其高表达与NSCLC患者预后不良显著相关。Further,CD151体外敲除抑制肿瘤增殖,迁移,和入侵。此外,与接种载体转染的肿瘤细胞的对照小鼠相比,用CD151过表达的肿瘤细胞接种的裸鼠表现出实质性的肿瘤增殖。值得注意的是,我们发现CD151的过表达通过与整合素相互作用而赋予细胞迁移和侵袭。我们接下来试图证明CD151通过激活NSCLC细胞中的EGFR/ErbB2调节下游信号传导途径。因此,我们推断CD151可能影响NSCLC对抗癌药物的敏感性.
结论:基于这些结果,我们通过靶向EGFR/ErbB2信号通路证明了CD151介导肿瘤进展的新机制,CD151促进NSCLC增殖,迁移,和入侵,这可能被认为是NSCLC治疗的潜在靶点。
BACKGROUND: Tetraspanins
CD151, a transmembrane 4 superfamily protein, has been identified participating in the initiation of a variety of cancers. However, the precise function of
CD151 in non-small cell lung cancer (NSCLC) remains unclear. Here, we addressed the pro-tumoral role of CD151 in NSCLC by targeting EGFR/ErbB2 which favors tumor proliferation, migration and invasion.
METHODS: First, the mRNA expression levels of CD151 in NSCLC tissues and cell lines were measured by RT-PCR. Meanwhile,
CD151 and its associated proteins were analyzed by western blotting. The expression levels of CD151 in NSCLC samples and its paired adjacent lung tissues were then verified by Immunohistochemistry. The protein interactions are evaluated by co-immunoprecipitation. Flow cytometry was applied to cell cycle analysis. CCK-8, EdU Incorporation, and clonogenic assays were used to analyze cell viability. Wound healing, transwell migration, and matrigel invasion assays were utilized to assess the motility of tumor cells. To investigate the role of
CD151 in vivo, lung carcinoma xenograft mouse model was applied.
RESULTS: High CD151 expression was identified in NSCLC tissues and cell lines, and its high expression was significantly associated with poor prognosis of NSCLC patients. Further, knockdown of
CD151 in vitro inhibited tumor proliferation, migration, and invasion. Besides, inoculation of nude mice with CD151-overexpressing tumor cells exhibited substantial tumor proliferation compared to that in control mice which inoculated with vector-transfected tumor cells. Noteworthy, we found that overexpression of CD151 conferred cell migration and invasion by interacting with integrins. We next sought to demonstrate that CD151 regulated downstream signaling pathways via activation of EGFR/ErbB2 in NSCLC cells. Therefore, we infer that CD151 probably affects the sensitivity of NSCLC in response to anti-cancer drugs.
CONCLUSIONS: Based on these results, we demonstrated a new mechanism of CD151-mediated tumor progression by targeting EGFR/ErbB2 signaling pathway, by which CD151 promotes NSCLC proliferation, migration, and invasion, which may considered as a potential target of NSCLC treatment.