Study on the effect of ectopic expression of LncRNA PVT1/EIF4A1 on the proliferation and migration of gastric cancer cells

Abstract

Objective Explore the effect of long non-coding RNA human plasmacytoma variant translocation 1 (PVT1) and human eukaryotic translation initiation factor4A1 (EIF4A1) on the proliferation and migration of gastric cancer (GC) cell SGC-7901. Methods Used Western Blot to detect EIF4A1 and real time-PCR to detect PVT1 in GC cells (SGC-7901, NCI-N87, MGC-803, BGC-823) and normal gastric mucosal cells (GES-1). Ectopic expressed PVT1 and EIF4A1 in GC cell SGC-7901 individually or together. Used Western Blot to detect the protein expression of EIF4A1. And used real time-PCR to detect the expression level of PVT1. Detected the migration ability of cells by the scratch test and the transwell assay. The proliferative ability of the cells was detected by the MTT and the clone formation assay. And the protein level of E-cadherin, N-cadherin, smad3 and TGF-β was detected by Western Blot. Results GC cells with low background expression of PVT1 and EIF4A1 were screened, and there was a statistical difference (P<0.05). After ectopic expression of PVT1 and EIF4A1 individually or jointly, the number of plate clone formation in the co-ectopic expression group was 492±8.72. The number of plate colonies formed in the PVT1 ectopic expression group alone was 251±1.53. And that in the EIF4A1 ectopic expression group and control group was 228±1.52 and 205±2.08 respectively. In MTT assay, the OD value of co-ectopic expression group was higher than that of individual ectopic expression group and control group. In the transwell assay, the number of migrated cells of the co-ectopic expression group was 308±29.10, and in the PVT1 ectopic expression group, EIF4A1 ectopic expression group and control group was 222±14.42, 206±10.58, and169±18.04 respectively. Cell scratch tests showed the wound healing rate in the co-ectopic expression group was higher than that in the individual ectopic expression group and the control group. The results of Western Blot showed that E-cadherin in the ectopic expression group was lower than that in the ectopic expression group and control group. N-cadherin, smad3 and TGF-β in the ectopic expression group were higher than those in the individual ectopic expression group and control group. All of these results were statistically significant (P<0.05). Conclusion Ectopic expression of PVT1 and EIF4A1 can promote the proliferation ability and migration ability of SGC-7901 cells by altering the expression of EMT related proteins.

Keywords

gastric cancer / PVT1 / EIF4A1 / cell proliferation / cell migration