沉默 SLC35F2 表达对肺癌 H1299 细胞行为的影响。
Influence on the behavior of lung cancer H1299 cells by silencing SLC35F2 expression.
机构信息
Department of Thoracic Surgery, Peking University People's Hospital, No 11, Xizhimen South, 100044 Beijing, China.
出版信息
Cancer Cell Int. 2013 Jul 23;13(1):73. doi: 10.1186/1475-2867-13-73.
BACKGROUND
To investigate the effects of RNA interference-mediated downregulation of Human Solute Carrier Family 35 member F2 (SLC35F2) expression on the biological behavior of lung cancer H1299 cells.
METHODS
The lentiviral vector of small interfering RNA targeting SLC35F2 was introduced into H1299 cells by liposome-mediated transfection. Expression of the SLC35F2 protein was measured by western blot. The proliferation of H1299 cells was determined by Cell Counting Kit-8 assay. The migration of H1299 cells was measured by Transwell migration assay. Cell cycle analysis used fluorescence-activated cell sorting.
RESULTS
SLC35F2 expression was markedly downregulated in H1299 cell clone (transfected with the lentiviral vector harboring small interfering RNA targeting SLC35F2). Proliferation decreased significantly compared with that of non-transfected H1299 cells. Transwell migration assay showed that fewer cells moved through the artificial basement membrane compared with untransfected H1299 cells (38.3 ± 5.7 vs. 113.5 ± 8.5, P < 0.05). The cell cycle of H1299 cells was changed, the percentage of H1299 cells in S and G2/M phases being significantly decreased compared with untransfected H1299 cells (S phase: 15.3% ± 3.0% vs. 27.0% ± 5.4%, P > 0.05; G2/M phase; 3.0% ± 1.1% vs. 10.5% ± 1.7%, P < 0.05), whereas the percentage of H1299 cells in G0/G1 phase increased markedly (81.7% ± 4.0% vs. 62.5% ± 1.9%, P < 0.05).
CONCLUSION
RNA interference-mediated downregulation of SLC35F2 expression by lentiviral vector can attenuate the proliferation, migration and invasion of H1299 cells.
背景
研究 RNA 干扰介导的人溶质载体家族 35 成员 F2（SLC35F2）表达下调对肺癌 H1299 细胞生物学行为的影响。
方法
通过脂质体介导转染将靶向 SLC35F2 的小干扰 RNA 的慢病毒载体导入 H1299 细胞。通过 Western blot 测定 SLC35F2 蛋白的表达。通过 Cell Counting Kit-8 测定 H1299 细胞的增殖。通过 Transwell 迁移测定测定 H1299 细胞的迁移。通过荧光激活细胞分选进行细胞周期分析。
结果
SLC35F2 表达在 H1299 细胞克隆（转染携带靶向 SLC35F2 的小干扰 RNA 的慢病毒载体）中明显下调。与未转染的 H1299 细胞相比，增殖显著降低。Transwell 迁移测定显示，与未转染的 H1299 细胞相比，穿过人工基底膜的细胞数量减少（38.3±5.7 对 113.5±8.5，P<0.05）。H1299 细胞的细胞周期发生改变，与未转染的 H1299 细胞相比，H1299 细胞 S 期和 G2/M 期的百分比明显降低（S 期：15.3%±3.0%对 27.0%±5.4%，P>0.05；G2/M 期：3.0%±1.1%对 10.5%±1.7%，P<0.05），而 G0/G1 期的 H1299 细胞百分比明显增加（81.7%±4.0%对 62.5%±1.9%，P<0.05）。
结论
慢病毒载体介导的 SLC35F2 表达下调通过 RNA 干扰可减弱 H1299 细胞的增殖、迁移和侵袭。
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