RNA interference targeting c-Met inhibits proliferation of human laryngeal carcinoma Hep-2 cell line in vitro and in its xenografts in nude mice
10.3760/cma.j.issn.1673-0860.2010.01.015
- VernacularTitle:RNA干扰抑制c-Met表达对喉癌Hep-2细胞体内外生长的影响
- Author:
Chang-You JI
1
;
Zhi-Nian XIE
;
Ji-Chuan CHEN
;
Yi-Nan WANG
;
Li-Lian GUAN
;
Hong-Tao LI
;
Min ZHANG
;
Rong-Rong LIU
Author Information
1. 第三军医大学附属大坪医院
- Keywords:
RNA interference;
Laryngeal neoplasms;
Cell line,tumor;
Proto-onengene proteins c-met
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2010;45(1):61-67
- CountryChina
- Language:Chinese
-
Abstract:
Objective The proto-oncogene c-Met was found to express on human laryngeal carcinoma Hep-2 cell line in previous research. In the present study, the author further examined whether inhibition of c-Met by RNA interference(RNAi) might inhibit biologic activity of Hep-2 cell line in vitro and proliferation using a murine laryngeal carcinoma model. Methods RNAi plasmid that can express small interfering RNA targeting c-Met or siRNA that did not match any known human coding mRNA( control siRNA plasmid) was designed,constructed,and transfected into Hep-2 cell line by using cationic liposome Lipofectamine2000 as transfecting agent. In vitro, the transfection efficacy was tested by RT-PCR and Western Blot method, then elected the most inhibitive c-Met-siRNA sequence. Cell proliferation, movement and invasion were studied using MTT, cell migration assay and cell invasion assay, respectively. The Hep-2 cells were transplanted into nude mice, then the time of tumor formation and growth were observed. After tumor formation, c-Met-siRNA was given as the anti-tumor therapy. Expression of c-Met, MMP-9 and VEGF were detected by Western Blot method. Results After the pSilencer2. 0/c-Met-shRNA recombinant plasmid transfeetion into laryngeal carcinoma Hep-2 cells, the expression of mRNA and proteinum of c-Met decreased significantly in Hep-2 cells. On the 35th day after tumor vaccination, the tumor volume was (138±27)mm~3 in c-Met-siRNA transfection group, Which was diminished significantly in contrast with control group( P<0. 01 ). The expression of c-Met, MMP-9 and VEGF in the tumor of experiment group was decreased significantly, respectively (P<0.05 ). Conclusions The results indicated that c-Met-siRNA can down-regulate the expression of c-Met and markedly inhibit laryngeal carcinoma Hep-2 cell proliferation, movement and invasion and the growth of transplantation tumor of nude mice. The siRNA expressing plasmid mediated gene therapy might be a new strategy in targeting molecular therapy of cancer of larynx.
