Expression of Spns2 in gastric cancer tissue and its effect on the proliferation and migration of gastric cancer cells / 医学研究生学报

ABSTRACT

Objective Few studies are reported on the correlation of Spinster homolog2(Spns2) with cancer.This study aims to investigate the expression of Spns2 in gastric cancer and the adjacent tissue and explore its effects on the proliferation, clone formation and migration of human gastric cancer cell line SGC-7901.Methods Samples of gastric cancer and the adjacent tissue were collected from 20 patients (12 males and 8 females) after surgery between February 2016 and August 2016. The expression of Spns2 in the gastric cancer and adjacent tissues was determined by immunohistochemistry. Eukaryotic expression vector pEX-1 (pGCMV/MCS/EGFP/Neo)-Spns2 and siRNA were constructed and transfected into the human gastric cancer cell line SGC-7901. The cells were divided into a pEX-1- Spns2 (siRNA-Spns2) group and a negative control pEX-1 (siRNA-NC) group. The mRNA and protein expressions of Spns2 were detected by RT-PCR and Western blot, respectively. And the effects of Spns2 on the migration, clone formation and proliferation of the SGC-7901 cells after Spns2 over-expressed and down-regulated were evaluated by cell counting kit-8 (CCK-8) assay, clone formation assay, transwell experiment and wound healing scratch assay.Results The expression of Spns2 was significantly lower in the gastric cancer than in the adjacent tissue (0.39±0.04 vs 0.45±0.02, P<0.05). After transfection, both Spns2 mRNA and protein levels were remarkably higher in the pEX-1-Spns2 (21.96±2.08 and 10.71±2.78) than in the pEX-1 group (0.88±0.12 and 0.83±0.16) (P<0.05), but markedly lower in the siRNA-Spns2 (0.35±0.07 and 0.53±0.07) than in the siRNA-NC group (0.91±0.09 and 0.92±0.08) (P<0.05). At 48, 72 and 96 hours, the proliferation of the SGC-7901 cells was significantly decreased in the pEX-1-Spns2 group as compared with the pEX-1 group (P<0.05) but increased in the siRNA-Spns2 group in comparison with the siRNA-NC group (P<0.05); the clone formation rate was reduced in the pEX-1-Spns2 group as compared with the pEX-1 group (\[33.30±3.81\]％ vs \[48.00±4.60\]％, P<0.05) but elevated in the siRNA-Spns2 group in comparison with the siRNA-NC group (\[48.38±4.22\]％ vs \[26.25±4.88\]％, P<0.05).Conclusion Spns2 is lowly expressed in gastric cancer, and its over-expression in SGC-7901 cells can reduce its proliferation, clone formation and migration. Spns2 may play a role in inhibiting the development and progression of gastric cancer.