Premium
Critical role for c‐FLIP L on Fas resistance in colon carcinoma cell line HT‐29
Author(s) -
Zang Fenglin,
Sun Baocun,
Zhao Xiulan,
Niu Ruifang,
Zhang Shiwu,
Yu Man,
Wei Xiyin,
Zhang Lin
Publication year - 2008
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/j.cellbi.2007.12.002
Subject(s) - flip , apoptosis , annexin , gene knockdown , microbiology and biotechnology , flow cytometry , small interfering rna , caspase 8 , cancer research , fas receptor , biology , cell culture , transfection , chemistry , programmed cell death , caspase 3 , biochemistry , genetics
The cellular Fas‐associated death domain‐like interleukin‐1β‐converting enzyme inhibitory protein‐long form (c‐FLIP L ) is a key regulator of Fas signaling, although owing a dominant‐negative homologue of caspase‐8, the role of c‐FLIP L remains controversial. In the present study, two pairs of small interfering RNA (siRNA) directed against c‐FLIP L were used to assess the effect of c‐FLIP L on Fas‐mediated apoptosis of colon carcinoma in vitro . HT‐29 cell line was selected for overexpression of c‐FLIP L and Fas with RT—PCR and flow cytometry analyses. After electroporation, the mRNA level of c‐FLIP L was significantly decreased (control siRNA versus c‐FLIP L siRNA, 77.97 ± 5.61% versus 26.22 ± 3.79%) and the maximum interfering efficiency was around 66.49% using semi‐quantitative RT—PCR analysis. Knockdown of c‐FLIP L with the specific siRNA sensitized colon carcinoma cells to Fas‐mediated apoptosis (control siRNA versus c‐FLIP L siRNA, 5.68 ± 2.11% versus 29.50 ± 2.27%) using DNA content analysis and Annexin V—FITC analysis. In conclusion, our study indicated that c‐FLIP L might be a suppressor of Fas‐mediated apoptosis in Fas antigen expressing colon carcinoma and therefore a potential target for novel anticancer therapies.