Mobile website
Basic Research
Effect of siRNA silencing EphB2 expression on proliferation and apoptosis of pancreatic cancer CFPAC-1 cells
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective : To investigate the effect of EphB2 on proliferation and apoptosis of human pancreatic cancer CFPAC-1 cells by target-silencing EphB2 expression using lentivirus-mediated RNA interference technique. Methods: The recombinant interfering plasmid targeting EphB2 gene was constructed and transfected into 293T cells by LipofectamineTM 2000, and the most effective interfering sequence was screened from inhibiting expression of EphB2 protein. The effective interference sequence was cloned into the pLentiGFP vector to construct lentivirus pLentiGFP-EphB2, and then transfected into CFPAC-1 cells. Stable cell line with down-regulated expression of EphB2 protein was obtained. Real-time PCR and Western blotting analysis were performed to determine the expression of EphB2 at mRNA and protein levels. Moreover, the effect of downregulation of EphB2 on proliferation and apoptosis of CFPAC-1 cells was detected by CCK8 and flow cytometry assay, respectively. Results: Recombinant lentivirus pLentiGFP-EphB2 was successfully constructed and stably transfected CFPAC-1 cells (CFPAC-1 EphB2 RNAi cells) were obtained. EphB2 expression at mRNA and protein levels in CFPAC-1 EphB2 RNAi cells were significantly lower than those at blank group, with silencing rate of EphB2 was 63% in the pLentiGFP-EphB2 group. In addition, the proliferation of CFPAC-1 EphB2 RNAi cells was increased more than those of the other two groups (1.89±0.17 vs 1.63±0.13, 1.71±0.22, P<0.05) and apoptosis was decreased \[(7.02±0.01)% vs (13.37±0.02)%, (15.71±0.02)%, P<0.05). Conclusion: pLentiGFP-EphB2 can effectively silence EphB2 expression in pancreatic cancer CFPAC-1 cells and then promote the proliferation and inhibit cell apoptosis.
Keywords:
Project Supported:
Project supported by the Natural Science Foundation of Tianjin (No. 09JCZDJC21800)
Clc Number:
