Chinese Clinical Oncology

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Targeted-regulation of MRAS expression by microRNA-148a-3p and its effects on invasion and migration of gastric cancer HGC-27 cells

WANG Jianguo,SUN Peida,LIU Haiyan,CHEN Lin.   

  1. Department of Interventional Radiology,Shuyang People’s Hospital of Jiangsu Province, Shuyang 223600,China
  • Received:2017-05-21 Revised:2017-08-22 Online:2017-10-30 Published:2017-10-30
  • Contact: CHEN Lin

Abstract: Objective To investigate the effect of microRNA-148a-3p(miR-148a-3p)on invasion, migration and expression of muscle RAS oncogene homolog(MRAS) in gastric cancer HGC-27 cells. Methods The cryopreserved gastric cancer cell line HGC-27 was recovered and cultured to the logarithmic growth phase. The miR-148a-3p analog and its negative control (miR-NC)were transfected into HGC-27 cells (miR-148a-3p transfection group and miR-NC group), and HGC-27 cells without transfection were chosen as blank control(untransfection group). The level of miR-148a-3p was detected by real-time quantitative PCR(QPCR) at 48 h after transfection. Transwell and scratch test were used to compare the number of penetrating-membrane cells and relative migration distance at 48 h after transfection in each group to evaluate the migration and invasion ability. The mRNA and protein levels of MRAS in each group at 48 h after transfection were detected by QPCR and Western blotting, respectively. Double luciferase reporter gene test was employed to verify the targeting regulation between miR-148a-3p and MRAS. Results QPCR detection showed that miR-148a-3p level of miR-148a-3p transfection group was 2.612±0.213,higher than 0.954±0.098 of untransfected group and 0.983±0.196 of miR-NC group,and the difference was statistically significant (P<0.05). Scratch results show that the relative migration distance of miR-148a-3p group was 0.615±0.019, lower than 1.021±0.019 of untransfection group and 0.948±0.022 of miR-NC group (P<0.05). Transwell assay showed that the transmembrane cell number in miR-148a-3p transfected group was 83±17, less than 124±17 of untransfection group and 136±26 of miR-NC group (P<0.05). QPCR and Western blotting results showed that mRNA and protein levels of MRAS in miR-148a-3p transfection group were 0.614±0.057 and 0.553±0.049, lower than 1.106±0.024 and 0.824±0.091 of untransfected group and 1.095±0.031 and 0.784±0.121 of miR-NC group (P<0.05). MiR-148a-3p significantly decreased the luciferase activity of wild-type MRAS-3’untranslated region plasmid transfected cells, but had no significant inhibitory effect on the luciferase activity of mutant MRAS plasmid transfected cells.Conclusion MiR-148a-3p may inhibit the invasion and migration of gastric cancer HGC-27 cells by targeting MRAS.

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