Silencing of SLC35F2 gene influences the biologic behavior of human lung cancer cell line H1299.

Abstract

e13512 Background: SLC35F2 is a member of solute carrier family 35 which encode nucleotide sugar transporters localizing in the Golgi apparatus and/or the endoplasmic reticulum, and transport nucleotide sugars that have pooled in the cytosol into the lumen of this organelle, where most of the glycoconjugate synthesis occurs. However, the SLC35F2 subfamily has not been completely characterized with regard to its biological activity. In this study, we tested the hypothesis that SLC35F2 influences the biological behavior of lung cancer cell line. We examined the effect of the inhibition of SLC35F2 expression on proliferation and invasion by establishing the SLC35F2 knockdown lung cancer cell line using lentivirus-mediated RNA interference (RNAi). Methods: The lentiviral vector of small interfering RNA targeting SLC35F2 was introduced into H1299 cells by liposome-mediated transfection. The expression of SLC35F2 protein was measured by Western blot. The proliferation of H1299 cells was determined by cell counting kit-8 (CCK-8) assay. The migration and invasion ability of H1299 cells was evaluated by Transwell migration and invasion assay. Cell cycle analysis was also performed by fluorescence-activated cell sorting (FACS). Results: SLC35F2 expression was markedly down-regulated in H1299 cell clone transfected with the lentiviral vector harboring small interfering RNA targeting SLC35F2. The proliferation of H1299 cells decreased significantly compared with untransfected H1299 cells. Transwell migration assay showed that less H1299 cells could move through the artificial basement membrane when 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. Compared with untransfected H1299 cells, the percentage of H1299 cells in S and G2/M phase decreased significantly (S phase: F15.31%±3.04% vs 27.00%±5.41%, P >0.05; GG2/M phase: F3.02%±1.10% vs 10.47%±1.72%, P <0.05), whereas the percentage of H1299 cells in G0/G1 phase increased remarkably (81.67%±3.98% vs 62.53%±1.89%, P <0.05). Conclusions: RNA interference-mediated down-regulation of SLC35F2 by lentiviral vector can attenuate the proliferation and migration of H1299 cells.