siRNA干扰VRK1表达对食管癌细胞BANF1蛋白表达及增殖迁移能力的抑制作用

耿杰; 李雨晴; 李晋; 王婷婷; 裴露; 刘红春

doi:10.3971/j.issn.1000-8578.2019.18.0941

siRNA干扰VRK1表达对食管癌细胞BANF1蛋白表达及增殖迁移能力的抑制作用

Down-regulation of VRK1 Expression by siRNA Suppresses BANF1 Expression and Proliferation and Migration Abilities of Esophageal Squamous Cell Carcinoma Cells

摘要

摘要:
目的探讨小干扰RNA干扰VRK1表达对食管癌细胞株BANF1蛋白的表达及其增殖迁移能力的影响。
方法将VRK1 siRNA转染EC109和EC1细胞系干扰VRK1的表达，Western blot法检测VRK1 siRNA干扰效率及siRNA干扰后食管癌细胞BANF1的表达。CCK-8实验和流式细胞术检测干扰VRK1表达后对细胞增殖能力的影响，Transwell细胞体外侵袭实验观察干扰前后细胞迁移能力的改变。
结果两株食管癌细胞VRK1 siRNA转染后，转染实验组VRK1蛋白表达分别下调62.40%和52.14%，同时BANF1蛋白表达分别下调24.51%和52.87%。转染12 h后，细胞增殖开始受到抑制，36 h抑制率最高，分别达19.41%、20.10%。VRK1和BANF1表达下调后，G₁、G₂期细胞比例下降，S期细胞比例上升。实验组细胞迁移个数下降，细胞迁移数和对照组相比差异有统计学意义（P < 0.05）。
结论 siRNA干扰VRK1表达后可降低食管鳞癌细胞BANF1蛋白的表达，使食管鳞癌细胞增殖周期阻滞在DNA合成期，分裂期细胞比例下降，抑制其增殖及迁移能力。

Abstract:
Objective To investigate the effect of VRK1 expression downregulated by siRNA on BANF1 expression and the proliferation and migration abilities of esophageal squamous cell carcinoma (ESCC) cells.
Methods VRK1 siRNA was transfected into EC109 and EC1 cell lines to interfere VRK1 expression. Western blot was used to detect the interference efficiency of VRK1 siRNA and BANF1 expression in esophageal carcinoma cells after siRNA interference. CCK-8 and flow cytometry were performed to detect the effect of VRK1 expression on cell proliferation. Transwell assay was used to observe the changes of cell migration ability.
Results After VRK1 siRNA transfection, VRK1 expression were down-regulated for 62.40% and 52.14%, and BANF1 protein expression were down-regulated significantly for 24.51% and 52.87% in EC109 and EC1 cell lines, respectively. The proliferation of EC109 and EC1 cell lines were decreased 12h after transfection, with the highest inhibition rate at 36h(19.41% and 20.10%). The cells percentages in G₂ and G₁ phases were decreased while that in S phase was increased after down-regulation of VRK1 and BANF1 expression. The number of cell migration was decreased in the experimental group, compared with control group (P < 0.01).
Conclusion Down-regulation of VRK1 expression by siRNA could inhibit the proliferation and migration abilities of esophageal squamous carcinoma cells, which may be achieved by down-regulating BANF1 expression.

HTML全文

参考文献(24)

施引文献

资源附件(0)