Document Detail


Effects of small interfering RNA targeting heparanase-1 combined with heparin on invasiveness of mouse hepatocellular carcinoma cell lines.
MedLine Citation:
PMID:  20800024     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND AND OBJECTIVE: Heparanase-1 (HPA-1) can promote angiogenesis and metastasis of malignant tumors and plays an important role in the genesis and development of tumors. This study was to explore the effects of specific small interfering RNA (siRNA) targeting HPA-1 combined with heparin on invasiveness of mouse hepatocellular carcinoma cells.
METHODS: The expression of HPA-1 in Hca-F, Hca-P, and Hepa1-6 cells, which have high, low, and no metastatic potential, respectively, was analyzed by reverse transcription-polymerase chain reaction (RT-PCR), Western blot analysis and enzyme-linked immunosorbent assay (ELISA). After transfection with two specific siRNAs targeting HPA-1, siRNA-1 and siRNA-2, and treatment with heparin, invasiveness of Hca-F cells was observed by Matrigel invasion assay.
RESULTS: HPA-1 was negative in Hepa1-6 cells while positive in both Hca-F and Hca-P cells. The expression levels of both HPA-1 mRNA and protein were obviously higher in Hca-F cells than in Hca-P cells. HPA-1 proteins could be secreted into culture supernatant of Hca-F and Hca-P cells, and the amount of secreted HPA-1 detected by Western blot analysis was larger in Hca-F cells than in Hca-P cells (1.34 ± 0.02 vs. 0.60 ± 0.01, P < 0.001), which was consistent with the results of ELISA. Both siRNA-1 and siRNA-2 downregulated the expression of HPA-1 and the siRNA-2 did more efficiently. The number of invasive Hca-F cells treated with siRNA-2 or heparin alone was larger than that of Hca-F cells treated with combination of them (9 ± 1 vs. 4 ± 1, P = 0.013; 15 ± 2 vs. 4 ± 1, P = 0.008), but smaller than that of untreated Hca-F cells (9 ± 1 vs. 22 ± 2, P = 0.006; 15 ± 2 vs. 22 ± 2, P = 0.026).
CONCLUSION: The combined application of specific siRNA targeting HPA-1 and heparin is more effective in inhibiting the invasiveness of mouse hepatoma cells.
Authors:
Sheng-Jin Yu; Xiao-Hui Kang; Jia-Ning Zhang; Hong-Mei Wang; Tao Xie; Wei Wang; Shu-Jing Wang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Chinese journal of cancer     Volume:  29     ISSN:  1000-467X     ISO Abbreviation:  Chin J Cancer     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-08-30     Completed Date:  2012-02-09     Revised Date:  2013-02-12    
Medline Journal Info:
Nlm Unique ID:  101498232     Medline TA:  Chin J Cancer     Country:  China    
Other Details:
Languages:  eng     Pagination:  816-23     Citation Subset:  IM    
Affiliation:
Department of Basic Medical Sciences, Medical College of Eastern Liaoning University, Dandong, Liaoning 118000, PR China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Line, Tumor
Cell Movement
Down-Regulation
Glucuronidase / biosynthesis*,  genetics,  secretion
Heparin / pharmacology*
Liver Neoplasms, Experimental / metabolism,  pathology*
Mice
Neoplasm Invasiveness*
Neoplasm Metastasis
RNA, Messenger / metabolism
RNA, Small Interfering / genetics*,  pharmacology
Transfection
Chemical
Reg. No./Substance:
0/RNA, Messenger; 0/RNA, Small Interfering; 9005-49-6/Heparin; EC 3.2.1.-/heparanase; EC 3.2.1.31/Glucuronidase

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


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