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EphA7基因沉默对人喉癌细胞系Hep-2和AMC-HN-8增殖、侵袭及凋亡的影响

Effect of EphA7 Silencing on Proliferation, Invasion and Apoptosis in Human Laryngeal Cancer Cell Lines Hep-2 and AMC-HN-8.

作者信息

Xiang Cheng, Lv Yuanjing, Wei Yanjie, Wei Jing, Miao Susheng, Mao Xionghui, Gu Xin, Song Kaibin, Jia Shenshan

机构信息

Department of Head and Neck Surgery, the Third Affiliated Hospital of Harbin Medical University, Harbin, China.

出版信息

Cell Physiol Biochem. 2015;36(2):435-45. doi: 10.1159/000430110. Epub 2015 May 11.

Abstract

AIMS

This study aimed to investigate the expression of EphA7 in human laryngeal squamous cell carcinoma (LSCC) tissues and disclose the potential roles and molecular mechanisms of EphA7 in LSCC.

METHODS

In the present study, we examined EphA7 expression and its function and mechanism in LSCC. EphA7 expression levels were investigated by quantitative real-time PCR (qRT-PCR), western blotting, and immunohistochemistry in a panel of 35 LSCC patient cases. To investigate the potential mechanism of EphA7 in human laryngeal cancer, we employed EphA7 siRNA to knockdown EphA7 expression in LSCC cell line Hep-2 and AMC-HN-8. Subsequently, MTT, TUNEL, qRT-PCR, and western blotting were performed to disclose the roles of EphA7 on proliferation, invasion and migration, and apoptosis in LSCC cell line Hep-2 and AMC-HN-8.

RESULTS

Depletion of EphA7 remarkably inhibited the proliferation and invasion of Hep-2 and AMC-HN-8 cells in comparison to control and EphA7 siRNA negative control (NC)-transfected cells. TUNEL staining assay demonstrated that, compared with the control group, the rate of apoptosis in the EphA7 siRNA group was significantly increased. In addition, knockdown of EphA7 in Hep-2 or AMC-HN-8 cells markedly decreased the expression of EphA7 and PTEN, which could contribute to apoptosis. However, the bpV(phen), a PTEN inhibitor, could attenuate anti-proliferation and pro-apoptotic effects of EphA7 siRNA in Hep-2 and AMC-HN-8 cells.

CONCLUSION

Up-regulation of EphA7 was observed in human LSCC samples and down-regulation of EphA7 effectively suppressed laryngeal carcinoma cell growth and promoted its apoptosis. Thus, EphA7 has a critical role in modulating cell growth and apoptosis, which serves as a potential therapeutic target in human LSCC.

摘要

目的

本研究旨在调查EphA7在人喉鳞状细胞癌(LSCC)组织中的表达,并揭示EphA7在LSCC中的潜在作用和分子机制。

方法

在本研究中,我们检测了EphA7在LSCC中的表达及其功能和机制。通过定量实时PCR(qRT-PCR)、蛋白质印迹法和免疫组织化学法对35例LSCC患者病例进行了EphA7表达水平的研究。为了研究EphA7在人喉癌中的潜在机制,我们使用EphA7 siRNA敲低LSCC细胞系Hep-2和AMC-HN-8中EphA7的表达。随后,进行MTT、TUNEL、qRT-PCR和蛋白质印迹法以揭示EphA7对LSCC细胞系Hep-2和AMC-HN-8增殖、侵袭和迁移以及凋亡的作用。

结果

与对照和EphA7 siRNA阴性对照(NC)转染细胞相比,EphA7的缺失显著抑制了Hep-2和AMC-HN-8细胞的增殖和侵袭。TUNEL染色分析表明,与对照组相比,EphA7 siRNA组的凋亡率显著增加。此外,在Hep-2或AMC-HN-8细胞中敲低EphA7显著降低了EphA7和PTEN的表达,这可能有助于凋亡。然而,PTEN抑制剂bpV(phen)可减弱EphA7 siRNA对Hep-2和AMC-HN-8细胞的抗增殖和促凋亡作用。

结论

在人LSCC样本中观察到EphA7上调,EphA7下调有效抑制喉癌细胞生长并促进其凋亡。因此,EphA7在调节细胞生长和凋亡中起关键作用,可作为人LSCC的潜在治疗靶点。

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