The Effect Of BOP1 On Invasion And Metastasis Of Gastric Cancer Cells And Its Mechanism

Objective:Gastric cancer is one of the most common digestive tract tumors in the world,and its morbidity and mortality rank fifth and third respectively in the world.The main cause of death in gastric cancer is the recurrence and metastasis of gastric cancer.In recent years,more and more literature reports that BOP1 is highly expressed in certain tumor tissues and promotes tumorigenesis and invasion and metastasis by affecting cell growth,proliferation and EMT.However,the role of BOP1 in gastric cancer has not been reported in the literature.This study mainly focused on gastric cancer cells AGS and BGC-823,and explored the expression of BOP1 in gastric cancer and its role and mechanism in the development,metastasis and invasion of gastric cancer.Methods:(1)To study the expression of BOP1 in gastric cancer,the expression levels of BOP1 mRNA and protein in serum and tissues of normal and gastric cancer patients and gastric cancer cells were detected by Elisa,RT-PCR or Western Blot.(2)The expression of BOP1 gene in gastric cancer cells was silenced by different siRNA sequences.According to the expression levels of BOP1 mRNA and protein in gastric cancer cells were detected by RT-PCR and Western Blot to selected the appropriate siRNA sequences.(3)To investigate the effect of BOP1 on the proliferation,migration and invasion of gastric cancer cells,siRNA was used to silence the expression of BOP1 gene in gastric cancer cells.CCK-8 assay,plate colony formation assay,wound healing assay and Transwell assay were used to test the ability of each group cells on proliferate,migrate and invasion.(4)Collect the transfected cells to study the molecular mechanism of BOP1 on gastric cancer cells and further study the effect of BOP1 on PI3K/AKT signaling pathway to detect the expression of PI3K/AKT signaling pathway-related proteins.(5)To investigate the effect of BOP1 gene on the expression of P53 gene and c-Myc gene in gastric cancer cells,the level of P53 and c-Myc protein in cells was detected by Western Blot after silencing the BOP1 gene in gastric cancer cells.Results:(1)Compared with normal human serum,gastric tissue and gastric mucosal epithelial cells,the expression levels of BOP1 mRNA and protein were up-regulated in serum,gastric cancer tissues and gastric cancer cells of gastric cancer patients.(2)Transfection results showed that in gastric cancer cells AGS and BGC-823,siRNA1 sequence and siRNA2 sequence were the lowest in BOP1 mRNA and protein after transfection into gastric cancer cells.The siRNA-1 sequence and the siRNA-2 sequence were selected and transfected into gastric cancer cells AGS and BGC-823 for subsequent experiments.(3)The results showed that in the CCK-8 method,the cell colony formation experiment,the wound healing experiment and the Transwell experiment,compared with BC group and si-NC,the proliferation,migration and invasion ability of si-BOP1 cells were significantly decreased.(4)Compared with adjacent tissues,the expression level of GSK3? protein in gastric cancer tissues of gastric cancer patients was significantly reduced.The expression of GSK3? was significantly increased in gastric cancer cells after knocking down the BOP1 gene.Compared with the BC group and the si-NC group,the total PI3 K and total AKT protein levels in the si-BOP1 group did not change significantly,and the p-PI3 K and p-AKT protein levels decreased significantly.(5)Compared with the BC group and the si-NC group,the expression level of P53 protein in the si-BOP1 group was significantly increased,and the expression level of c-Myc was significantly decreased.Conclusion:Compared with normal human serum,gastric cancer tissue and gastric cancer cells,the expression of BOP1 in serum,gastric cancer and gastric cancer cells of gastric cancer patients was significantly increased.It suggests that BOP1 is closely related to the occurrence and development of gastric cancer.BOP1 can promote the proliferation,migration and invasion of gastric cancer by using siRNA knockout technology,CCK-8 assay,plate colony formation assay,wound healing assay and Transwell experimental study.And BOP1 may affect the proliferation,migration and invasion of gastric cancer cells through PI3K/AKT/GSK3? signaling pathway.Compared with the BC group and the si-NC group,the c-Myc protein of the proto-oncogene was significantly decreased and the expression of the tumor suppressor gene P53 protein was significantly increased in the si-BOP1 group.It suggests that BOP1 may also promote the development of gastric cancer by affecting the expression of oncogenes and tumor suppressor genes.