Effects of lentivirus-mediated epidermal growth factor-like domain 7 silencing on proliferation and invasion of human laryngeal carcinoma Hep-2 cells
10.3760/cma.j.issn.1673-0860.2014.08.006
- VernacularTitle:慢病毒介导的类表皮生长因子域7基因沉默抑制喉癌Hep-2细胞增殖和侵袭的研究
- Author:
Jingjia LI
1
;
Jin YE
;
Shimin ZHUANG
;
Tao WANG
;
Zhiyuan WANG
;
Lihong CHANG
;
Gehua ZHANG
Author Information
1. 中山大学附属第三医院耳鼻咽喉头颈外科
- Keywords:
RNA,small interfering;
Laryngeal neoplasms;
Epidermal growth factor;
Cell proliferation;
Neoplasms invasiveness;
Lentivirus;
Tumor cells,cultured
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2014;49(8):643-648
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects of epidermal growth factor-like domain 7 (EGFL7) gene silencing on the proliferation and invasion ablity of laryngeal carcinoma cells.Methods A lentiviral vector expressing EGFL7 shRNA was constructed and transfected into human laryngeal carcinoma Hep-2 cells.The expressions of EGFL7 mRNA and protein were detected by Real-time PCR and Western blot,respectively.Cell proliferation was evaluated by CCK-8 assay,cell cycle and apoptosis were tested by flow cytometry,and cell invasion was detected by transwell invasion assay.Results The relative expression level s of EGFL7 mRNA and protein in EGFL7-SuRNA group were svgnificantly lower than control group (P ＜0.001).Western blot analysis proved that the relative expression of EGFL7 protein in NC group,Lenti-NCgroup and Lenti-EGFL7 group was (0.39 ±0.12),(0.36 ±0.14) and (0.07 ±0.04),respectively.EGFL7 expression in Lenri-EGFL7 group was significantly inhibited than NC group (P ＜ 0.001),which confirmed that the recombinant lentivirus was successfully transfected into Hep-2 cells.The proliferation of Hep-2 cells was significantly inhibited after transfection (P ＜ 0.01).Compared with the NC group and Lenti-NC group,the proportion of cells in S phase was significantly increased in Lenti-EGFL7 group (P ＜ 0.01),and the proportion in G1 phase was significantly decreased (P ＜ 0.05).Cell apoptosis assay showed that the apoptotic rate in Lenti-EGFL7 group (66.2 ± 1.28) ％ was significantly increased in NC group (6.09-±3.28)％ and Lenti-NC group (9.86 ±2.13)％.In Transwell invision assay,the mean number of cells coming through the Metrigel in Lenti-EGFL7 group was significantly decreased than in the NC group and Lenti-NC group (P ＜ 0.01).Conclusion The proliferation and invasion ablity of laryngeal carcinoma Hep-2 cells can be inhibited by siRNA mediated EGFL7 gene silencing.
