Niu Juntao, Huang Yongwang, Zhang Lun
Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy Tianjin 300060, China ; Department of Otorhinolaryngology, The Second Hospital of Tianjing Medical University Tianjin 300211, China.
Department of Otorhinolaryngology, The Second Hospital of Tianjing Medical University Tianjin 300211, China.
Int J Clin Exp Pathol. 2015 Jun 1;8(6):6255-61. eCollection 2015.
CXCR4 has been reported in various types of human cancer, which is associated with cancer progression and metastasis. However, the investigation of CXCR4 in laryngeal cancer is extremely rare. In the present study, we used lentivirus-mediated shRNA targeting CXCR4 to silenced CXCR4 expression in Hep-2 cells and evaluated the effect of long-term suppression of CXCR4 on Hep-2 growth and metastasis. The Cell proliferation was analyzed by MTS assay, and the invasion and metastasis potentials were analyzed using wound healing and transwell assays, respectively. Our results showed that lentivirus-mediated shRNA effectively infected Hep-2 cells and suppressed CXCR4 expression, and inhibited cell growth of Hep-2 cells. Cell invasion and apoptosis were decreased concomitantly with the reduction in CXCR4 protein expression. Further analysis revealed that CXCR4 silencing caused the reducion of CXCR4, CXCL12, TIMP2, VEGF and MMP9, and the phosphorylation levels of IκB, AKT and MAPK, and also decreased the activity of NF-κB. These results suggested that knockdown of CXCR4 inhibits the invasion and metastasis of Hep-2 through PI3K/AKT and MAPK signaling pathways, by decreasing NF-κB activities to down-regulate VEGF, TIMP-2 and MMP-9 expression. These data demonstrate that the inhibition of CXCR4 may be an effective interventional therapeutic strategy in laryngeal cancer.
CXCR4已在多种人类癌症中被报道,它与癌症进展和转移相关。然而,关于CXCR4在喉癌中的研究极为罕见。在本研究中,我们使用慢病毒介导的靶向CXCR4的短发夹RNA(shRNA)来沉默Hep-2细胞中CXCR4的表达,并评估长期抑制CXCR4对Hep-2细胞生长和转移的影响。通过MTS法分析细胞增殖,分别使用伤口愈合试验和Transwell试验分析侵袭和转移潜能。我们的结果表明,慢病毒介导的shRNA有效感染Hep-2细胞并抑制CXCR4表达,进而抑制Hep-2细胞的生长。细胞侵袭和凋亡随着CXCR4蛋白表达的降低而减少。进一步分析显示,CXCR4沉默导致CXCR4、CXCL12、TIMP2、VEGF和MMP9减少,IκB、AKT和MAPK的磷酸化水平降低,同时NF-κB的活性也降低。这些结果表明,敲低CXCR4通过PI3K/AKT和MAPK信号通路抑制Hep-2细胞的侵袭和转移,通过降低NF-κB活性来下调VEGF、TIMP-2和MMP-9的表达。这些数据表明,抑制CXCR4可能是喉癌一种有效的干预治疗策略。