骨桥蛋白在人喉鳞状细胞癌及其裸鼠异种移植模型中的功能特性

Functional characterization of OPN in human laryngeal squamous cell carcinoma and its xenograft model in nude mice.

作者信息

Chen Jianqiu, Zhou Qi, Zhu Chunsheng, Zhu Minhui, Tian Yongsheng, Li Guojun, Tao Xiaofeng, Zheng Hongliang

机构信息

Department of Otolaryngology Head and Neck Surgery, General Hospital of Jinan Military Region Jinan 250031, Shandong Province, PR China.

Department of Tumor Biological Treatment, The Third Affiliated Hospital, Soochow University Changzhou 213003, Jiangsu Province, China.

出版信息

Int J Clin Exp Med. 2015 Jan 15;8(1):164-72. eCollection 2015.

PMID:25784985

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4358440/

Abstract

BACKGROUND

Osteopontin (OPN) is involved in promotion of cancer cells by regulating various facets of tumor progression such as cell proliferation, angiogenesis and metastasis. To understand the role of OPN in laryngeal squamous cell carcinoma (LSCC), we thus explored the biological function of OPN in LSCC after silencing OPN expression by RNA interference (RNAi).

METHOD

The OPN expression in tumor tissues of LSCC was determined immunohistochemically in both LSCC and adjacent normal tissues. Lentivirus vector with RNAi small hairpin gene sequence of OPN (named LV-shOPN) was transfected into Hep-2 cells and transplanted into BALB/c-nu mice. After siRNA transfection, the viability of Hep-2 cells was examined by MTS, OPN expression was detected by Western blotting, and tumor angiogenesis was assessed by microvessel densities (MVD).

RESULTS

The difference of positive rate of OPN in 72 cases LSCC (54 cases, 75.0%) and adjacent normal tissues (15 cases, 20.8%) was statistically significant (P<0.001) and the OPN expression was also significantly correlated with tumor stage, grade and the presence of lymph node. Hep-2 cells infected with LV-shOPN significantly decreased OPN expression, in comparison to cells with LV-shNon transfection (as the control) (P<0.05). The constructed LV-shOPN effectively inhibited the viability of Hep-2 cell and growth of xenograft tumors in nude mice (all P<0.050). The expression of OPN and MVD was significantly decreased in xenograft tumors (all P<0.05).

CONCLUSION

RNAi silencing of OPN expression can significantly inhibit tumor growth and angiogenesis of Hep-2 cells, and OPN may be considered as one of gene targeting therapy for LSCC.

摘要

背景

骨桥蛋白（OPN）通过调节肿瘤进展的各个方面，如细胞增殖、血管生成和转移，参与促进癌细胞生长。为了解OPN在喉鳞状细胞癌（LSCC）中的作用，我们通过RNA干扰（RNAi）沉默OPN表达后，探讨了OPN在LSCC中的生物学功能。

方法

采用免疫组织化学方法检测LSCC肿瘤组织及癌旁正常组织中OPN的表达。将携带OPN的RNAi小发夹基因序列的慢病毒载体（命名为LV-shOPN）转染至Hep-2细胞，并接种于BALB/c-nu小鼠。siRNA转染后，采用MTS法检测Hep-2细胞活力，Western印迹法检测OPN表达，微血管密度（MVD）评估肿瘤血管生成。

结果

72例LSCC组织中OPN阳性率为75.0%（54例），癌旁正常组织中OPN阳性率为20.8%（15例），差异有统计学意义（P<0.001），且OPN表达与肿瘤分期、分级及淋巴结转移显著相关。与转染LV-shNon（作为对照）的细胞相比，感染LV-shOPN的Hep-2细胞OPN表达显著降低（P<0.05）。构建的LV-shOPN有效抑制了Hep-2细胞活力及裸鼠移植瘤生长（均P<0.05）。移植瘤中OPN表达及MVD均显著降低（均P<0.05）。