Mobile website
Archive > Volume 20 Issue 6 > 2013,20(6):685-689. DOI:10.3872/j.issn.1007-385X.2013.06.009 Prev Next
Basic Research
Effect of down-regulation of the Slug gene expression by RNAi on cell cycle, proliferation and invasion of lung cancer A549 cells
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective:To study the effect of silencing Slug gene on proliferation, cell cycle and invasion ability of lung cancer A549 cells by RNA interference technique. Methods: The recombinant plasmid expressing short hairpin RNA (shRNA) targeting Slug gene was constructed and named as pGPU6-GFP-Neo-Slug. A549 cells were transfected with pGPU6-GFP-Neo-Slug and the negative control plasmid pNeg-shRNA with liposome method. Real-time PCR and Western blotting analysis was performed to determine the expression of Slug mRNA and protein after transfection, respectively. The proliferation, cell cycle distribution and cell invasion of A549 cells were detected by CCK-8 assay, flow cytometry and transwell assay, respectively. Results: pGPU6-GFP-Neo-Slug vector was successfully constructed and transfected into A549 cells with transfection rate of nearly 90%. Compared to the control and pNeg-shRNA group, Slug mRNA (\[0.23±0.01\] vs \[0.97±0.08\], \[1.0±0.09\]; all P＜0.05) and protein (\[0.20±0.09\] vs \[1.0±0.32\], \[1.13±0.26\]; all P＜0.05) expression level was significantly reduced in pSlug-shRNA group. Compared to the control and pNeg-shRNA group, the inhibition rate of proliferation in Slug-silencing A549 cells was significantly increased (\[35.3±5.4\]% vs \[15±02%\], \[3.3±0.7%\]; all P< 0.05); the cell multiplication index was significantly decreased(\[32.92±069\] vs \[48.19±0.71\], \[42.88±0.75\]; all P<0.05); the cell number in G1 phase was significantly increased(\[67.08±092\] vs \[52.81±0.78\], \[56.12±0.73\]; all P<0.05); and the cell invasion ability of A549 was significantly reduced (number of alive A549 cells through the matrigel chamber：\[55±9\] vs \[169±12\], \[173±15\]; all P<0.01). Conclusion: pGPU6-GFP-Neo-Slug vector transfected into lung cancer A549 cell can effectively silence Slug gene expression, inhibit cell proliferation, influence cell cycle and inhibit cell invasion of A549 cells.
Keywords:
Project Supported:
Project supported by the National Natural Science Foundation of China（No.81102057）
Clc Number:
