Abstract
Objective: To investigate the effect of stable transfection with NAD+-linked 15-hydroxyprostaglandin dehydrogenase (15-PGDH) on the growth, proliferation and migration of gastric cancer cells. Methods: The levels of 15-PGDH in four kinds of gastric cancer cells with different differentiation rates were compared by RT-PCR measurements, and the poorly differentiated gastric cancer cells SGC7901 were chose to perform follow-up experiments. SGC7901 cells were transfected with recombinant plasmid pcDNA3/15-PGDH and empty plasmid pcDNA3 as a control by using lipofectamine 2000. Cells with steady expression capability were sorted out by G418. RT-PCR and Western blot were used to confirm the transfection and expression of 15-PGDH in SGC7901 cells. MTT, cell scratch assay, soft-agar colony formation assays were used to determine the proliferation, migration and cell clone formation activities of SGC7901 with stable transfection of 15-PGDH. Flow cytometry assay was used to examine the effect of 15-PGDH on cell cycle and apoptosis. RT-PCR was used to analyze the levels of cell cycle-related genes (p53, p21 and p16) and apoptosis-related genes (Survivin, BCL-2 and Caspase3). Results: Both recombinant plasmid pcDNA3/15-PGDH and empty vector plasmid pcDNA3 were successfully transfected into human gastric cancer SGC7901 cell line. Proliferation, migration and cell clone formation capabilities of pcDNA3/15-PGDH groups were significantly inhibited compared with other two control groups (P<0.05). Flow cytometry results demonstrated an increased fraction of sub-G1 phase and the increase of apoptotic cells for pcDNA3/15-PGDH groups compared with other two control groups (P<0.05). The levels of p16, p21 and p53 mRNA for pcDNA3/15-PGDH groups were higher than those in other two control groups (p<0.05). pcDNA3/15-PGDH groups exhibited a higher level of caspase3 mRNA but lower levels of survivin and BCL-2 mRNA in SGC7901 cells compared with other two control groups (P< 0.05). Conclusion: The gene transfection of 15-PGDH has negative effects on the growth, proliferation and migration of gastric cancer SGC7901 cells by inducing apoptosis and cell cycle arrest.
Highlights
Gastric cancer is a commonly observed malignant tumor and the most common cause of cancer-related mortality worldwide
Recent studies have shown that the reduction or loss of 15-hydroxyprostaglandin dehydrogenase (15-PGDH) expression may lead to tumor progression. 15PGDH expression is significantly low in many malignant tumors such as colon cancer, non-small cell lung cancer, breast carcinoma, esophageal squamous cell carcinoma and adenocarcinoma, gastric cancer [2,3,4,5,6,7,8] and some precancerous lesions, such as atrophic gastritis [6]. 15-PGDH was significantly down regulated in H.pylori–infected gastric human tissues and it may be a useful marker and a potential therapeutic target in H. pylori–induced gastric carcinogenesis [7]
We constructed a SGC7901 cell line with stable expression of 15-PGDH gene and investigated the effect of 15-PGDH on the growth, proliferation and migration of gastric cancer, and we examined the association of 15-PGDH with gastric cancer cell
Summary
Gastric cancer is a commonly observed malignant tumor and the most common cause of cancer-related mortality worldwide. Our previus studies showed that 15-PGDH inhibitors could promote the proliferation of human gastric cancer cells and COX-2 inhibitors could inhibit the growth of gastric cancer cells by promoting the expression of 15-PGDH [11,12]. We constructed a SGC7901 cell line with stable expression of 15-PGDH gene and investigated the effect of 15-PGDH on the growth, proliferation and migration of gastric cancer, and we examined the association of 15-PGDH with gastric cancer cell
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
