Effect of shRNA-mediated silence of H-ras gene on proliferation of human SACC-M cells / 中华口腔医学杂志
Chinese Journal of Stomatology
;
(12): 113-117, 2008.
Article
in Chinese
| WPRIM
| ID: wpr-235971
ABSTRACT
<p><b>OBJECTIVE</b>To examine the effects of H-ras gene silence on cell cycle, proliferation and apoptosis of salivary adenoid cystic carcinoma -M (SACC-M) cell lines.</p><p><b>METHODS</b>The plasmid H-ras-shRNA, containing the sequence of shRNA targeting H-ras, and HK-shRNA (without interfering effect) were constructed and transfected into SACC-M cells. The cell line with shRNA plasmid stable expression was isolated by G418. The expression levels of H-ras were detected by RT-PCR and protein immunofluorescent assay; cell cycle and cell apoptosis were analyzed by flow cytometry (FCM). The proliferation of cell was also determined by subcutaneous tumor formation in nude mice.</p><p><b>RESULTS</b>After transfection of H-ras-shRNA plasmid, the mRNA expression of H-ras in SACC-M cells was down-regulated by 61.80% and protein expression of H-ras was inhibited by 62.76%; the cell proliferation was inhibited obviously; the G0G1 phase cells were increased. The cell apoptosis rate of H-ras-shRNA group was significantly higher than that of HK-shRNA group (P <0.05). The volume of subcutaneous tumor in nude mice was significantly smaller in Hras-shRNA group than in control group.</p><p><b>CONCLUSIONS</b>The recombinant plasmid HRAS-shRNA could efficiently down-regulate the expression of H-ras gene and protein, induce apoptosis of SACC-M cells and simultaneously inhibit proliferation of these cells in vitro and in vivo.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Salivary Gland Neoplasms
/
Transfection
/
Proto-Oncogene Proteins p21(ras)
/
Apoptosis
/
Carcinoma, Adenoid Cystic
/
Gene Silencing
/
RNA, Small Interfering
/
Cell Line, Tumor
/
Cell Proliferation
Limits:
Animals
/
Female
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Stomatology
Year:
2008
Type:
Article
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