Suppression of Aurora-A by RNA interference inhibits laryngeal cancer Hep-2 cell growth.
- Author:
Hao ZHANG
1
;
Xue-hua CHEN
;
Chang-ping CAI
;
Shi-li WANG
;
Bing-ya LIU
;
Liang ZHOU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Aurora Kinase A; Aurora Kinases; Carcinoma, Squamous Cell; genetics; metabolism; pathology; Cell Line, Tumor; Cell Transformation, Neoplastic; Focal Adhesion Protein-Tyrosine Kinases; metabolism; Gene Silencing; Humans; Laryngeal Neoplasms; genetics; metabolism; pathology; Matrix Metalloproteinase 2; metabolism; Mice; Mice, Nude; Protein-Serine-Threonine Kinases; genetics; RNA Interference; RNA, Small Interfering; genetics; Transfection
- From: Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2012;47(1):58-63
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effects of knockdown of Aurora-A by RNA interference on laryngeal cancer Hep-2 cell growth in vitro and in vivo.
METHODSA plasmid containing siRNA against Aurora-A was constructed and transfected into human laryngeal cancer cell line Hep-2. Measurements included the CCK-8 assay for viability and proliferation, Transwell assay for invasion, colony formation assay for cell anchorage-independent growth. Western blot and immunohistochemistry assay for protein expression. Tumorigenicity was observed in vivo.
RESULTSIn Hep-2 cells transfected by Aurora-A siRNA (designated as siRNA-3), protein expression of Aurora-A was suppressed by 52%. In CCK-8 assay, absorbance value of siRNA-3 cells (3.268 ± 0.106, (x(-) ± s)) was lower than that of Hep-2 cells (3.722 ± 0.152, F = 17.634, P < 0.001). In Transwell assay, the average invasive cells per field in siRNA-3 cells (110.0 ± 18.0) was less than that in Hep-2 cells (236.0 ± 26.0, F = 26.462, P < 0.01). In colony formation assay, the average colony number of siRNA-3 cells (31.0 ± 6.6) was lower than that of Hep-2 cells (104.0 ± 14.0). The average tumor size in siRNA-3 group was (127.77 ± 174.83) mm(3), which was less than Hep-2 cell group (837.26 ± 101.80) mm(3), (F = 28.187, P < 0.001). Silencing of Aurora-A decreased the expression of focal adhesion kinase (FAK) and matrix metalloproteinase-2 (MMP-2), key regulators in cell adhesion and invasion.
CONCLUSIONSThe knockdown of Aurora-A inhibits the growth and invasiveness of Hep-2 cells in vitro and in vivo, which may be a promising therapeutic strategy for LSCC.
