Document Detail


Combining siRNAs at two different sites in the EGFR to suppress its expression, induce apoptosis, and enhance 5-fluorouracil sensitivity of colon cancer cells.
MedLine Citation:
PMID:  17169374     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: The epidermal growth factor receptor (EGFR) has played an important role in the growth and apoptosis of colon cancer. The RNA interference (RNAi) technique can suppress gene expression, but the effects of double combining sites RNAi targeting EGFR have not been well understood. METHODS: pU6-EGFR-shRNA-1 and pU6-EGFR-shRNA-2 expressive vectors were transfected to the LoVo cells. Five groups were selected for the study: Group 1, the control cells; group 2, the negative control plasmid vector HK; group 3, pU6-EGFR-shRNA-1; group 4, pU6-EGFR-shRNA-2; group 5, pU6-EGFR-shRNA-1 and pU6-EGFR-shRNA-2, half for each. The mRNA and protein expression were assessed by Real Time quantitative PCR and Western blot. Apoptosis was determined via flow cytometry. IC(50) and the inhibition ratio of 5-fluorouracil (5-FU) were carried out by CCK-8. RESULTS: In groups 3, 4, and 5, the mRNA expression was decreased by (80.22 +/- 3.42)%, (81.30 +/- 2.83)%, and (90.58 +/- 2.76)%, respectively, and the protein expression was decreased by (74.11 +/- 4.02)%, (73.39 +/- 2.30)%, and (90.39 +/- 3.34)%, respectively. Meanwhile, the cell apoptosis increased by (10.43 +/- 0.49)%, (10.13 +/- 0.39)%, and (14.17 +/- 0.53)%, respectively. The IC(50) of 5-FU and cell inhibition ratio analysis demonstrated that there were significant differences between the following three: group 5, groups 3 and 4, and groups 1 and 2. CONCLUSIONS: Both pU6-EGFR-shRNA-1 and pU6-EGFR-shRNA-2 are capable of suppressing EGFR expression of the LoVo cell and can promote apoptosis and increase the cell toxicity of 5-FU. The double combining sites RNAi technique is significantly better than a single site.
Authors:
Xiangbai Wu; Yongzhi Deng; Guobin Wang; Kaixiong Tao
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-12-12
Journal Detail:
Title:  The Journal of surgical research     Volume:  138     ISSN:  0022-4804     ISO Abbreviation:  J. Surg. Res.     Publication Date:  2007 Mar 
Date Detail:
Created Date:  2007-02-16     Completed Date:  2007-04-05     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  0376340     Medline TA:  J Surg Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  56-63     Citation Subset:  IM    
Affiliation:
Department of General Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
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MeSH Terms
Descriptor/Qualifier:
Antimetabolites, Antineoplastic / pharmacology*
Apoptosis / drug effects,  physiology
Blotting, Western
Cell Line, Tumor
Colonic Neoplasms / drug therapy*
Combined Modality Therapy / methods
Drug Resistance, Neoplasm / genetics
Flow Cytometry
Fluorouracil / pharmacology*
Gene Expression Regulation, Neoplastic / drug effects,  genetics
Gene Therapy / methods*
Humans
Plasmids / genetics
RNA, Small Interfering* / genetics
Receptor, Epidermal Growth Factor / genetics*,  metabolism
Reverse Transcriptase Polymerase Chain Reaction
Transfection
Chemical
Reg. No./Substance:
0/Antimetabolites, Antineoplastic; 0/RNA, Small Interfering; 51-21-8/Fluorouracil; EC 2.7.10.1/Receptor, Epidermal Growth Factor

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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