Effect of E2F1 Knockdown on proliferation and invasion of clear cell renal cell carcinoma cell line Caki-2
Yu Gao, Fan Yang, Weihao Chen, Qingbo Huang, Qing Ai, Dong Ni, Hanxin Wu, Xu Zhang
Abstract
Yu Gao, Fan Yang, Weihao Chen, Qingbo Huang, Qing Ai, Dong Ni, Hanxin Wu, Xu Zhang
Abstract
Objective To study the effect of E2F1 knockdown on proliferation,migration and invasion of clear cell renal cell carcinoma cell line Caki-2.Methods Small interfering RNA (siRNA)-E2F1and negative control sequences were transfected into Caki-2 cells by Lipo2000.The E2F1 expression levels were detected by using real-time quantitative polymerase chain reaction (PCR) and Western blotting respectively after transfection.MTS and Transwell assays were used to detect variations in cell proliferation,migration and invasion.Results Compared to untransfected group (130.100 ± 6.301) × 10-5 and negative control group (134.600 ± 7.163) × 10-5,the mRNA levels in siRNA-E2F1 group (37.770 ± 3.782) × 10-5were significantly reduced (P <0.01).The protein levels were consistent with mRNA levels.MTS assay demonstrated the absorbance in siRNA-E2F1 group was significantly decreased at the time points of 24,48,72 and 96 h after transfection (P < 0.05),and Transwell assay revealed that the number of migrating cells in siRNA-E2F1 group (38.330 ±4.041) was significantly reduced at 48 h after transfection as compared with untransfected group (83.000 ± 8.888) and negative control group (87.330 ± 6.429) (P <0.01),and the number of invasive cells in siRNA-E2F1 group (78.330 ±7.572) was strongly reduced at 48 h after transfection as compared with untransfected group (143.30 ± 10.066) and negative control group (147.000 ± 14.000) (P <0.01).Conclusion Knockdown of E2F1 significantly decreased proliferation,migration and invasion of Caki-2 cells. Key words: Clear cell renal cell carcinoma; E2F1 gene ; Proliferation; Migration; Invasion
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Objective To study the effect of E2F1 knockdown on proliferation,migration and invasion of clear cell renal cell carcinoma cell line Caki-2.Methods Small interfering RNA (siRNA)-E2F1and negative control sequences were transfected into Caki-2 cells by Lipo2000.The E2F1 expression levels were detected by using real-time quantitative polymerase chain reaction (PCR) and Western blotting respectively after transfection.MTS and Transwell assays were used to detect variations in cell proliferation,migration and invasion.Results Compared to untransfected group (130.100 ± 6.301) × 10-5 and negative control group (134.600 ± 7.163) × 10-5,the mRNA levels in siRNA-E2F1 group (37.770 ± 3.782) × 10-5were significantly reduced (P <0.01).The protein levels were consistent with mRNA levels.MTS assay demonstrated the absorbance in siRNA-E2F1 group was significantly decreased at the time points of 24,48,72 and 96 h after transfection (P < 0.05),and Transwell assay revealed that the number of migrating cells in siRNA-E2F1 group (38.330 ±4.041) was significantly reduced at 48 h after transfection as compared with untransfected group (83.000 ± 8.888) and negative control group (87.330 ± 6.429) (P <0.01),and the number of invasive cells in siRNA-E2F1 group (78.330 ±7.572) was strongly reduced at 48 h after transfection as compared with untransfected group (143.30 ± 10.066) and negative control group (147.000 ± 14.000) (P <0.01).Conclusion Knockdown of E2F1 significantly decreased proliferation,migration and invasion of Caki-2 cells. Key words: Clear cell renal cell carcinoma; E2F1 gene ; Proliferation; Migration; Invasion
Key concepts: Gene knockdown, Transfection, Cell growth, Small interfering RNA, E2F1, Cell culture, Molecular biology, Cell