Effects of RNA interference targeting epidermal growth factor receptor on the apoptosis in and chemosensitivity of a human cutaneous carcinoma cell line Colo-16
10.3760/cma.j.issn.0412-4030.2011.04.012
- VernacularTitle:RNA干扰抑制表皮生长因子受体表达对Colo-16细胞凋亡及化疗敏感性的影响
- Author:
Hui WANG
;
Zhiping WEI
;
Yanqun LIU
- Publication Type:Journal Article
- Keywords:
RNA interference;
Receptor,epidermal growth factor,Carcinoma,squamous cell;
Sirolimus
- From:
Chinese Journal of Dermatology
2011;44(4):263-266
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of a short hairpin RNA targeting epidermal growth factor recereceptor (EGFR-shRNA) on Colo-16 cell apoptosis and sensitivity to rapamycin. Methods The expression vector of EGFR-specific shRNA was constructed. Colo-16 cells were classified into 4 groups, normal control group remaining untreated, liposome group transfected with lipofectamine 2000, negative control group transfected with shRNA-NC/Iipofectamine 2000 and positive interference group transfected with the expression vector of shRNA-EGFR/Lipofectamine 2000. After additional culture, immunocytochemistry and Western blot were conducted to detect the protein expression of EGFR, and flow cytometry to measure the apoptosis in Colo-16cells. MTT assay was performed to measure the sensitivity of Colo-16 cells to rapamycin. Results Compared with the normal control group, the expression of EGFR was down-regulated by 43.3% in positive interference group (F= 44.6, P< 0.05), and the sensitivity to rapamycin was increased by 2.44 folds. The apoptosis rate in positive interference group was (12.65±0.091)%, significantly different from that in the normal control group (F = 2042.9, P < 0.05). Conclusion The plasmid expression vector containing shRNA targeting EGFR can effectively suppress the expression of EGFR by Colo-16 cells, enhance the sensitivity of Colo-16 cells to rapamycin and induce the apoptosis in Colo-16 cells.
