Document Detail


Knockdown of survivin gene expression by RNAi induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721.
MedLine Citation:
PMID:  15655839     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
AIM: To investigate the survivin gene expression in human hepatocellular carcinoma cell line SMMC-7721 and the effects of survivin gene RNA interference (RNAi) on cell apoptosis and biological behaviors of SMMC-7721 cells. METHODS: Eukaryotic expression vector of survivin gene RNAi and recombinant plasmid pSuppressorNeo-survivin (pSuNeo-SVV), were constructed by ligating into the vector, pSuppressorNeo (pSuNeo) digested with restriction enzymes Xba I and Sal I and the designed double-chain RNAi primers. A cell model of SMMC-7721 after treatment with RNAi was prepared by transfecting SMMC-7721 cells with the lipofectin transfection method. Strept-avidin-biotin-complex (SABC) immunohistochemical staining and RT-PCR were used to detect survivin gene expressions in SMMC-7721 cells. Flow cytometry was used for the cell cycle analysis. Transmission electron microscopy was performed to determine whether RNAi induced cell apoptosis, and the method of measuring the cell growth curve was utilized to study the growth of SMMC-7721 cells before and after treatment with RNAi. RESULTS: The eukaryotic expression vector of survivin gene RNAi and pSuNeo-SVV, were constructed successfully. The expression level of survivin gene in SMMC-7721 cells was observed. After the treatment of RNAi, the expression of survivin gene in SMMC-7721 cells was almost absent, apoptosis index was increased by 15.6%, and the number of cells was decreased in G2/M phase and the cell growth was inhibited. CONCLUSION: RNAi can exert a knockdown of survivin gene expression in SMMC-7721 cells, and induce apoptosis and inhibit the growth of carcinoma cells.
Authors:
Sheng-Quan Cheng; Wen-Liang Wang; Wei Yan; Qing-Long Li; Li Wang; Wen-Yong Wang
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Publication Detail:
Type:  In Vitro; Journal Article    
Journal Detail:
Title:  World journal of gastroenterology : WJG     Volume:  11     ISSN:  1007-9327     ISO Abbreviation:  World J. Gastroenterol.     Publication Date:  2005 Feb 
Date Detail:
Created Date:  2005-01-18     Completed Date:  2005-03-14     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  100883448     Medline TA:  World J Gastroenterol     Country:  China    
Other Details:
Languages:  eng     Pagination:  756-9     Citation Subset:  IM    
Affiliation:
Department of Pathology, Faculty of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032, Shaanxi Province, China.
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MeSH Terms
Descriptor/Qualifier:
Apoptosis
Carcinoma, Hepatocellular / physiopathology,  therapy*
Cell Division
Cell Line, Tumor / physiology,  ultrastructure
Flow Cytometry
Gene Expression
Gene Therapy / methods*
Humans
Liver Neoplasms / physiopathology,  therapy*
Microscopy, Electron, Transmission
Microtubule-Associated Proteins / genetics*
Neoplasm Proteins
RNA, Small Interfering*
Restriction Mapping
Chemical
Reg. No./Substance:
0/BIRC5 protein, human; 0/Microtubule-Associated Proteins; 0/Neoplasm Proteins; 0/RNA, Small Interfering

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


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