| Objective:The third leading cause of cancer-related death is gastric cancer.Compared with the clinical status of patients with advanced disease with poor long-term survival rate,some progress made in the treatment of gastric cancer is insignificant.At the same time,although great progress has been made in surgery and adjuvant therapy,the 5-year survival rate of gastric cancer patients is still low due to the recurrence of metastatic cancer.Therefore,elucidating the molecular mechanisms of gastric cancer metastasis is crucial for improving patient survival.Studies have shown that EGFR family members such as Erb B1 and Erb B3 are involved in tumor progression and metastasis.In addition,the co-overexpression of Erb B1 and Erb B3 in gastric cancer is often associated with poor prognosis,and at the same time,they can be used for targeted therapy of gastric cancer patients.Gene-targeted therapy has become a research hotspot in recent years.With the in-depth research on gastric cancer proto-oncogenes and tumor suppressor genes in various fields,many gene therapy targets for gastric cancer have been discovered.Ebp1 is a newly discovered intracellular binding protein of Erb B3.Previous studies have found that Ebp1 can induce the proliferation of gastric cancer cells,but whether the role of gastric cancer cell migration and invasion is related to Ebp1 is still unclear.For this reason,we carried out this study to explore the correlation between malignant behavior of gastric cancer and Ebp1,as well as the underlying molecular mechanism of the interaction,and to provide experimental evidence for targeted therapy and clinical diagnosis and treatment of gastric cancer cells.Method:The ualcan software was used to analyze the expression of PA2G4 gene in various cancers and its differential expression between gastric cancer tissues and adjacent tissues.Then,the transfection of recombinant lentivirus was used to knock down the expression of Ebp1 in gastric cancer cells.The migration and invasion abilities of cells were detected by scratch assay and Transwell assay.Ebp1 and EMT protein network,Ebp1 and PI3K/Akt/m TOR protein network were constructed using STRING software.The effect of Ebp1 on the expression of EMT-related proteins and the effect of Ebp1 on the phosphorylation levels of key proteins in the PI3K/Akt/m TOR signaling pathway were detected by Western blot.Result:1.The results of ualcan database showed that abnormal expression of Ebp1 protein was observed in several cancer tissues.The expression level of Ebp1 in gastric cancer tissue and its adjacent tissue was significantly different,and the expression of Ebp1 in paracancerous tissue was much lower than that in gastric cancer tissue(P <0.05).2.Detection of transfection effect: Using Western blot detection,it was found that there were significant differences in the expression level of Ebp1 protein.Compared with the sh-NC group,the protein expression level of sh-Ebp1 was significantly decreased(***P<0.001),and the results indicated that the MGC-803 gastric cancer cell line had been successfully constructed after knocking down the Ebp1 gene.3.Scratch test: the difference in the scratch healing rate is particularly obvious between the sh-Ebp1 group and sh-NC.The statistical analysis results show that the scratch healing rate shows a cliff-like decline in the sh-Ebp1 group(24h: **P<0.01,48h: ***P<0.001),the results show that the migration ability of gastric cancer MGC-803 cells is closely related to the Ebp1 gene,that is,with the knockout of the Ebp1 gene,the migration ability of cancer cells appears significantly suppressed phenomenon.4.Transwell experiments showed that the number of cells permeating the filtration membrane was significantly different between the sh-Ebp1 group and the sh-NC phase,and the number of cells permeating the filtration membrane in the sh-Ebp1 group was significantly reduced(***P < 0.001),the results showed that after the Ebp1 gene was silenced,the invasion ability of MGC-803 gastric cancer cells was greatly reduced.5.The network of Ebp1 and EMT-related proteins was constructed using STRING software,and the results were analyzed: Ebp1 and EMT proteins were closely related.Statistical analysis of the protein level detection results of Western blot showed that with the change of the variable Ebp1 gene knockdown,the protein expression also changed,such as the expression of E-cadherin protein was significantly increased(***P<0.001);N-cadherin protein,Vimentin protein,Snail protein,Slug protein and MMP2 protein were significantly decreased(***P<0.001).6.In order to find the relationship between Ebp1 and PI3K/Akt/m-TOR signaling pathway,we used STRING software to construct the protein network of the two.Through the observation and analysis of the protein network,we found that the relationship between Ebp1 and PI3K/Akt/m TOR signaling pathway proteins close.Therefore,using Western blot to detect the protein level showed that the protein expression levels of p-PI3 K,p-AKT,and p-m TOR were significantly different between the Control group and the LY294002 group,and the expression levels of these three proteins in the LY294002 group decreased.(***P<0.001),while the protein expression level of Ebp1 did not change significantly;the protein expression levels of p-AKT and p-m TOR in the Perifosine group were significantly decreased compared with those in the Control group(***P<0.001).),the protein expression levels of Ebp1 and p-PI3 K did not change significantly;compared with the Control group,the protein expression of p-m TOR decreased in the rapamycin group(**P<0.01).Compared with the Control group,the expression levels of PI3 K and p-AKT;Sh-Ebp1 decreased,including: Ebp1,p-PI3 K,p-AKT and p-m TOR proteins.7.The results of scratch test and Transwell test showed that compared with the control group,the ability of gastric cancer cells to migrate and invade changed with the treatment.The LY294002 group,the Perifosine group,the rapamycin group and the sh-Ebp1 group showed specific performance,that is,the migration and invasion abilities of these groups were significantly inhibited(scratch: **P<0.01,Transwell :***P<0.001).Statistical analysis of the results of Western blot with software showed that: with the Control group as the reference,the expression level of E-cadherin protein was up-regulated(***P<0.001)in the following groups:LY294002 group,Perifosine group,rapamycin group after treatment At the same time,the expression levels of these proteins were down-regulated(***P<0.001):N-cadherin,Vimentin,Snail,Slug,MMP2 proteins.Conclusion:1.After knocking down the Ebp1 gene,the migration and invasion ability of gastric cancer MGC-803 cells was weakened.2.After knocking down the Ebp1 gene,the EMT process of MGC-803 gastric cancer cells can be inhibited.3.Ebp1 can regulate the EMT process through the PI3K/Akt/m TOR pathway,and promote the migration and invasion of gastric cancer MGC-803 cells. 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