| Background and Aims:Gastric cancer is one of the most common malignant tumors in the world.In 2012,there were 950,000 new cases of gastric cancer worldwide,and 723,000 deaths.More than 40 percent of them are in China.Because gastric cancer is not easy to be detected in the early stage,it is often in the middle and late stage when diagnosed,and there is no effective treatment method,gastric cancer is still an important disease that seriously affects people’s life and health.Therefore,active exploration of the pathogenesis of gastric cancer and search for possible drug targets for the treatment of gastric cancer will provide important ideas and theoretical basis for the diagnosis and treatment of gastric cancer.GFR alpha 2 expression was significantly decreased in gastric cancer tissues.Glial cell line-derived neurotrophic factor family receptor alpha 2is a member of the Glial cell line-derived neurotrophic factor family receptor alpha family,which plays an important role in the normal development of kidneys and nerves.In addition to kidney and nervous system development,GFR alpha 2 also plays an important role in many biological processes.GFR alpha 2 is a marker of first and second cardiac field cardiac progenitor cells(CPs)that has been used for CPs isolation..GFRA2 is a marker of early B cells,and single cell PCR analysis suggested it combined with lymphocyte antigen 6 family member D(Ly6D)and bone marrow stromal cell antigen 1(BST1)to define a development trajectory linking classical common lymphoid progenitors to CD19~+progenitors。Previous studies have found that overexpression of GFR alpha2 level in pancreatic cancer tissues is positively correlated with tumor size and poor prognosis,promoting pancreatic cancer growth and increasing chemotherapy resistance.In acute myeloid leukemia(AML)cells,artemin(ARTN)/GFRα3 and ARTN/GFRα2 ligand/coreceptor complexes activate ret proto-oncogene(RET)signaling to promote leukemogenesis.However,related pathways can inhibit tumor development in colorectal cancer.Moreover,the role and mechanism of GFRalpha 2 in the development of gastric cancer have not been reported.This project aims to explore the role and mechanism of GFRalpha 2 in the development of gastric cancer..However,its role and mechanism in the development of gastric cancer have not been reported.The purpose of this study is to explore the role and mechanism of GFR alpha 2 in the development of gastric cancer.To solve this scientific problem,the following three parts are proposed:(1)to explore the expression of GFR alpha 2 in gastric cancer;(2)to clarify the specific expression of GFR alpha 2 in gastric cancer and the location in the gastric cancer cells through the gastric cancer tissue chip.Western blot was used to further verify its expression in gastric cancer cell lines.Immunohistochemistry was used to detect the localization of GFR alpha 2 in gastric cancer cells.to explore the GFR alpha 2 in gastric cancer cell life activities:on the one hand,through a small interference RNA silencing technology knockdowning GFR alpha 2 expression in gastric cancer cell lines,on the other hand,through the plasmid expressing GFR alpha 2,after the above processing cell to understand gastric cancer cell life activities associated with the subsequent changes,through CCK8 proliferation and clone formation research,flow cytometry research cycle and apoptosis.to explore the detailed molecular mechanism of GFR alpha 2’s role in gastric cancer:on the one hand,through literature review,it was found that GFR alpha 2 could bind to neuturin and transmembrane protein tyrosine kinase RET,and the expression of neuturin and RET in gastric cancer was understood by western blotting method to determine whether they were upstream and downstream molecules of GFR alpha 2.Methods:GFR alpha 2 expression in gastric cancer tissues and cytoplasmic localization were determined by gastric cancer microarray.Its expression in gastric cancer cell lines was further verified by western blot.Immunohistochemistry was used to detect the nucleoplasma localization of GFR alpha 2 in gastric cancer cells.The effects of GFR alpha2 overexpression plasmid and GFR alpha 2 interference on cell proliferation and apoptosis in gastric cancer cell lines SGC7901 and MGC803 were analyzed.The expression of neuturin and transmembrane protein tyrosine kinase RET in gastric cancer tissues was studied by western blot method to determine whether they were upstream and downstream of GFR alpha 2.GFR alpha 2 overexpression plasmid transfected with gastric cancer cell lines SGC7901 and MGC803 was analyzed.Results:The results showed that GFR alpha 2 was significantly decreased in gastric cancer tissues and cell lines.The microarray test of gastric cancer showed significant difference between the positive expression of GFR alpha 2 and paracancer cytoplasm and membrane,with paracancer higher than cancer,p<0.01.Overexpression of GFR alpha 2 inhibited proliferation of gastric cancer cells,while knockdown of GFR alpha 2promoted proliferation of gastric cancer cells.Neuturin had low expression in gastric cancer tissues and high expression in the corresponding paracancer tissues.The above differences were statistically significant(P<0.05)and the expression trend was consistent with that of GFRα2 in gastric cancer.After transfection with GFRα2overexpression plasmid,the expression of Ret in gastric cancer cell lines MGC803 and SGC7901 was significantly up-regulated compared with the control group,and the above differences were statistically significant(P<0.05).It is consistent with the expression trend of GFRα2.We speculate that Neuturin and Ret are involved in the molecular mechanism of GFRα2 in gastric cancer.Conclusions:The expression of Glial cell line-derived neurotrophic factor family receptorα2(GFRα2)was significantly reduced in gastric cancer cell lines and gastric cancer tissues,suggesting that GFRα2 plays a role in the occurrence and development of gastric cancer and is of certain significance to the study of gastric cancer.The overexpression of GFRα2significantly inhibit the proliferation of gastric cancer cells,while knocking down GFRα2 significantly promote the proliferation of gastric cancer cells.GFRα2 acts as a tumor suppressor gene by inhibiting the proliferation of gastric cancer cells.We speculated that neuturin was the upstream of the GFRα2,and Ret was the downstream of GFRα2.The results show that the expression of neuturin was low in gastric cancer tissue and high in the corresponding adjacent tissue,and the above differences were statistically significant(P<0.05),and the expression trend was consistent with that of GFRα2 in gastric cancer tissues.After transfection with GFRα2 overexpression plasmid,Ret expression in gastric cancer cell lines MGC803 and SGC7901 was significantly upregulated compared with the vector group,and the above differences were statistically significant(P<0.05).It is consistent with the expression trend of GFRα2. |
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