Effects of damage-regulated autophagy regulator gene on the SGC7901 human gastric cancer cell line

Abstract

The aim of this study was to investigate the effects of the adenoviral-mediated autophagy gene, damage-regulated autophagy regulator (DRAM), on the proliferation and autophagy of SGC7901 human gastric cancer cells in vitro. The recombinant adenovirus, AdMax-pDC315-DRAM-EGFP, working as a virus vector of DRAM was constructed and infected into the SGC7901 human gastric cancer cell line. The MTT assay was used to determine the growth rate of the SGC7901 cells. Activation of autophagy was monitored with monodansylcadaverin (MDC) staining following AdMax-pDC315-DRAM-EGFP treatment. Immunofluorescent staining was used to examine the expression of microtubule-associated protein 1 light chain 3 (LC3), and western blotting was used to examine the expression of apoptosis- and autophagy-associated proteins, including Beclin1, p53, p21 and B-cell lymphoma 2 (Bcl-2), in the culture supernatant. The viability of the SGC7901 cells was activated by AdMax-pDC315-DRAM-EGFP treatment. The AdMax-pDC315-DRAM-EGFP-treated cells exhibited positive LC3 expression detected by immunoreactivity and MDC staining. Inductions in the expression of the apoptosis-related proteins, p53 and p21, and the autophagic protein, Beclin1, were revealed by western blot analysis. By contrast, downregulation of the apoptosis-related protein, Bcl-2, following AdMax-pDC315-DRAM-EGFP treatment was identified. In conclusion, the present study demonstrated that AdMax-pDC315-DRAM-EGFP treatment resulted in upregulation of the level of autophagy and induction of cell proliferation in the SGC7901 human gastric cancer cell line in vitro.