The Role And Mechanism Of ABL2 In Gastric Cancer

Objective: To detect the difference expression of ABL2 in gastric cancer tissues and corresponding adjacent non-cancerous tissues?normal gastric mucosal epithelial cells and gastric cancer cell lines.To explore its role in the development of gastric cancer and its molecular mechanism,hope to provide new ideas and therapy of gastric cancer.Methods: Public database was used to predict the expression of ABL2 mRNA levels in the tissues of gastric cancer and corresponding adjacent non-cancerous tissues,and the relationship between the overall survival rate of patients with gastric cancer and the expression of ABL2 was investigated.The expression of ABL2 mRNA and protein levels in gastric cancer tissues and adjacent non-cancerous were detected by immunohistochemistry(IHC)and fluorescence quantitative PCR(qRT-PCR).The expression of ABL2 in normal gastric mucosal epithelial cells and gastric cancer cell lines was detected by Western blotting.Lentiviral vectors packed small interfering RNA(si RNA)and Lentiviral Activation Particles(sc-417779)were used to knock down and overexpress ABL2 in MGC-803 and BGC-823 gastric cancer cell lines then verified their efficiency.Transwell migration experiment and wound healing experiment were performed to analyze the effect of ABL2 on the migration ability of gastric cancer cells.The effect of ABL2 on the proliferation of gastric cancer cells was observed by plate clone formation test and Cell Counting Kit-8(CCK-8)assay.Western blotting detected the change of the proteins associated with EMT?metastasis and proliferation,further analysis of the expression of TGF-? and the Hippo signaling pathway related expression,preliminarily analyze its role in the development of gastric cancer and its mechanism.To obtain the detailed role of ABL2 in gastric cancer,the stability of the lower expression ABL2 in MGC-803 cells to nude mice subcutaneous tumor-burdended experiment,about 20 days,the subcutaneous tumors was taken to measure its weight and volume,and the effect of ABL2 on subcutaneous tumors was analyzed.Results: 1.qRT-PCR results showed that compared with corresponding adjacent non-cancerous tissues,ABL2 mRNA level in gastric cancer tissues was higher(61.7%,29/47),the results consist with tumor public databases.IHC results found the expression of ABL2 in gastric cancer tissue increased and the corresponding adjacent non-cancerous tissues in low expression.Western blotting detected the protein level of ABL2 and found it both expressed in normal gastric mucosal epithelial cells and gastric cancer cell lines and its expression was higher in gastric cancer cells,in addition to the low expression of ABL2 in BGC-823 cells,it was highly expressed in the rest of the gastric cancer cell lines.High expression of ABL2 in gastric cancer patients led to their overall survival rate low than low expression of ABL2 in gastric cancer patients.2.Transwell migration experiment? wound healing experiment? plate clone formation test and Cell Counting Kit-8(CCK-8)assay all showed that inhibited the expression of ABL2 in MGC-803 cells could decrease the ability of cell migration and proliferation.The expression of the epithelial cell marker E-cadherin was increased,and the expression of the mesenchymal cell markers,such as Snail?N-cadherin and Vimentin,were decreased,and the EMT was inhibited.It can also increase the expression of TIMP1 and TIMP2 while decrease MMP2 and MMP9.What is more,the expression of PCNA decreased and inhibited the growth of tumor cells.Also,the expression of Bcl-XL and caspase3 was reduced,the expression of Bax,Cleaved caspase3,Cleaved caspase9 and Cleaved PARP increased,promoting apoptosis of gastric cancer cells.The nude mice subcutaneous tumor-burdended experiment found that knockdown of ABL2 in MGC-803 could inhibit the proliferation of tumor cells.We also found that TGF-? and Hippo signaling pathway play a significant role in it.3.ARG Lentiviral Activation Particles infected BGC-823 cells to overexpress ABL2 in it,we found it could increase the migration and proliferation ability of gastric cancer cells,also,inhibit apoptosis in gastric cancer cells.The related protein levels also changed correspondingly.Conclusion: ABL2 plays an important role in the development of gastric cancer and may induce the metastasis and proliferation while inhibit apoptosis of gastric cancer cells by regulating the TGF-? and Hippo signaling pathway.ABL2 is expected to be a new target for the diagnosis of gastric cancer and new ideas for the treatment of gastric cancer.