Effect of shRNA-mediated silence of H-ras gene on proliferation of human SACC-M cells

• Published in: Zhonghua kouqiang yixue zazhi Vol. 43; no. 2; p. 113
• Main Authors: Yu, Li-Jie, Wang, Jie, Dong, Fu-Sheng, Shi, Hong, Li, He-Xiang, Gu, Hong-Tao
• Format: Journal Article
• Language: Chinese
• Published: China 01.02.2008
• Subjects: Animals; Apoptosis; Carcinoma, Adenoid Cystic - genetics; Carcinoma, Adenoid Cystic - pathology; Cell Line, Tumor; Cell Proliferation; Female; Gene Silencing; Humans; Mice; Mice, Inbred BALB C; Mice, Nude; Proto-Oncogene Proteins p21(ras) - genetics; RNA, Small Interfering - genetics; Salivary Gland Neoplasms - genetics; Salivary Gland Neoplasms - pathology; Transfection
• ISSN: 1002-0098

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. 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. 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 o