Effect of down-regulation of Notch2 expression on proliferation of esophageal squamous cell carcinoma cell line Eca109 through transfection with short-hairpin RNA
Author(s): WANG Xu-hui, HUA Sheng-hao, DENG Min, XIAO Li-hong, ZHOU Xing, ZHOU Rong, HU Juan, CAO Zhi-qin, LIU Qiang, JIAO Zhi-jun, Department of Laboratory Medicine, People’s Hospital of Xiangxiang, Department of Laboratory Medicine, Affiliated Children’s Hospital of Soochow University, Key Laboratory of Medical Immunology, Department of Laboratory Medicine, Affiliated Hospital of Jiangsu University
Pages: 501-506+540
Year: 2014 Issue: 6
Journal: Tumor
Keyword: Esophageal neoplasms; Receptor; Notch2; RNA interference; Cell proliferation; Cell cycle;
Abstract: Objective: To investigate the effect of blocking Notch2 signaling pathway on the proliferation of esophageal squamous cell carcinoma cell line Eca109 through down-regulating the expression level of Notch2 by transfection with short-hairpin RNA(shRNA). Methods: The expression of Notch receptor in multiple tumor cell lines including esophageal squamous cell carcinoma Eca109 cells was detected by real-time fl uorescence quantitative-PCR(RFQ-PCR). Then Eca109 cells with high expression of Notch2 were transfected with Control-shRNA and specific shRNA of Notch2(Notch2-shRNA), respectively. The mRNA expressions of Notch2 and its downstream gene Hes-1 in Eca109 cells after transfection with shRNA were detected by RFQ-PCR. The Notch2 protein expression was detected by Western blotting. The proliferation of Eca109 cells was detected by cell counting kit-8(CCK-8). In additon, fl ow cytometry was performed to assess the cell cycle distribution of Eca109 cells. Results: Notch2 gene highly expressed in Eca109 cell line. Compared with the control group, the expression levels of Notch2 mRNA and protein and its downstream gene Hes-1 were significantly decreased after transfection with Notch2-shRNA(P < 0.01). Moreover, the proliferation of Eca109 cells transfected with Notch2-shRNA for 72 h was inhibited(P < 0.05), and the percentage of Eca109 cells in G0/G1 phase in Notch2-shRNA transfection group(70.66±6.25)% was significantly higher as compared with Control-shRNA transfection group [(45.34±1.88)%] and the untransfection group [(47.10±1.70)%](P < 0.01). Conclusion: Notch2 is significantly highly expressed in Eca109 cells in vitro. The down-regulation of Notch2 expression can suppress the proliferation of Eca109 cells through inducing cell cycle arrest in the G0/G1 phase.
Pages: 501-506+540
Year: 2014 Issue: 6
Journal: Tumor
Keyword: Esophageal neoplasms; Receptor; Notch2; RNA interference; Cell proliferation; Cell cycle;
Abstract: Objective: To investigate the effect of blocking Notch2 signaling pathway on the proliferation of esophageal squamous cell carcinoma cell line Eca109 through down-regulating the expression level of Notch2 by transfection with short-hairpin RNA(shRNA). Methods: The expression of Notch receptor in multiple tumor cell lines including esophageal squamous cell carcinoma Eca109 cells was detected by real-time fl uorescence quantitative-PCR(RFQ-PCR). Then Eca109 cells with high expression of Notch2 were transfected with Control-shRNA and specific shRNA of Notch2(Notch2-shRNA), respectively. The mRNA expressions of Notch2 and its downstream gene Hes-1 in Eca109 cells after transfection with shRNA were detected by RFQ-PCR. The Notch2 protein expression was detected by Western blotting. The proliferation of Eca109 cells was detected by cell counting kit-8(CCK-8). In additon, fl ow cytometry was performed to assess the cell cycle distribution of Eca109 cells. Results: Notch2 gene highly expressed in Eca109 cell line. Compared with the control group, the expression levels of Notch2 mRNA and protein and its downstream gene Hes-1 were significantly decreased after transfection with Notch2-shRNA(P < 0.01). Moreover, the proliferation of Eca109 cells transfected with Notch2-shRNA for 72 h was inhibited(P < 0.05), and the percentage of Eca109 cells in G0/G1 phase in Notch2-shRNA transfection group(70.66±6.25)% was significantly higher as compared with Control-shRNA transfection group [(45.34±1.88)%] and the untransfection group [(47.10±1.70)%](P < 0.01). Conclusion: Notch2 is significantly highly expressed in Eca109 cells in vitro. The down-regulation of Notch2 expression can suppress the proliferation of Eca109 cells through inducing cell cycle arrest in the G0/G1 phase.
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