Effects of lentivirus-mediated epidermal growth factor-like domain 7 silencing on proliferation and invasion of human laryngeal carcinoma Hep-2 cells.
- Author:
Jingjia LI
1
;
Jin YE
1
;
Shimin ZHUANG
1
;
Tao WANG
1
;
Zhiyuan WANG
1
;
Lihong CHANG
1
;
Gehua ZHANG
2
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Cell Cycle; Cell Division; Cell Line, Tumor; Cell Proliferation; EGF Family of Proteins; Gene Silencing; Genetic Vectors; Hep G2 Cells; Humans; Laryngeal Neoplasms; metabolism; virology; Lentivirus; RNA, Messenger; RNA, Small Interfering; Transfection
- From: Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2014;49(8):643-648
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the effects of epidermal growth factor-like domain 7 (EGFL7) gene silencing on the proliferation and invasion ablity of laryngeal carcinoma cells.
METHODSA 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.
RESULTSThe 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-NC group 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).
CONCLUSIONThe proliferation and invasion ablity of laryngeal carcinoma Hep-2 cells can be inhibited by siRNA mediated EGFL7 gene silencing.