Mechanism Of RAD54B Regulating The Biological Behavior Of Hepatoma Cells By Inhibiting DUSP10 To Activate MAPK Signaling Pathway

Research BackgroundPrimary liver cancer is one of the highly malignant gastrointestinal tumors.In2018,the global incidence of liver cancer ranked sixth in malignant tumors and fourth in cancer-related deaths.The main pathological types of primary liver cancer include hepatocellular carcinoma,intrahepatic cholangiocarcinoma,and mixed types.Hepatocellular carcinoma is the most common subtype,accounting for about 80% of total cases.In recent years,research has found that DNA damage repair is closely related to tumorigenesis.Failure to properly repair DNA damage in mammals can lead to the development of various tumors,and abnormal activation of the DNA damage repair(DDR)system can also promote tumorigenesis and progression..Studies have shown that RAD54 B,as a novel DNA damage repair protein,is abnormally expressed in many malignancies.Previous research by our research group found that the high expression of RAD54 B in liver cancer tissues was positively correlated with poor prognosis of liver cancer patients,and promoted the proliferation and inhibition of apoptosis of liver cancer cells.However,the molecular mechanism by which RAD54 B affects the biological behavior of liver cancer cells remains unclear.This paper intends to construct a stable interference cell model of RAD54 B,use microarray and bioinformatics to screen out the key molecules that mediate RAD54 B to promote the malignant biological behavior of liver cancer cells,and analyze the signal pathways that the latter may regulate.Research methodHuh-7 and SNU-449 liver cancer cell lines were selected for the construction of RAD54 B lentivirus stable interference cell models.MTT experiments,wound healing experiments,Transwell experiments,and flow cytometry were used to detect RAD54 B knockdown.Effects of invasion and apoptosis.In RAD54 B stably interfered liver cancer cells,microarray was used to detect differentially expressed genes before and after RAD54 B knockdown,and GO and KEGG enrichment analysis were performed to screen out key molecules regulated by RAD54 B.QRT-PCR and WesternBlot were used for further verification.SiRNA technology was used to further transiently silence key molecules in hepatoma cells stably interfered with RAD54 B.The transfection efficiency was verified by qRT-PCR and Western Blot,and the effect on the molecular expression of downstream signaling pathways was detected.MTT experiments,wound healing experiments,Transwell experiments,and flow cytometry were used to detect the effects of silencing of key molecules on the biological behavior of liver cancer cells stably interfering with RAD54 B.Research result1.Knocking down RAD54 B of liver cancer cells can reduce the proliferation,migration and invasion of liver cancer cells,and promote apoptosis of liver cancer cells.2.Using microarray and bioinformatics analysis,we found that DUSP10 expression was up-regulated after knocking down liver cancer cell RAD54 B.Further,qRT-PCR and Western Blot were used to identify DUSP10 as a key molecule regulated by RAD54 B.3.In liver cancer cells stably interfered by RAD54 B,the expression of DUSP10 can be effectively silenced by siRNA technology.4.In RAD54 B stably interfered liver cancer cells,silencing the expression of DUSP10 enhances the proliferation,migration and invasion of liver cancer cells,and inhibits apoptosis of liver cancer cells.5.In RAD54 B stably interfered liver cancer cells,silencing DUSP10 expression affects the phosphorylation level of key molecules in the MAPK signaling pathway.Analysis conclusionRAD54B promotes the proliferation,migration and invasion of liver cancer cells by inhibiting the expression of DUP10,and inhibits apoptosis of liver cancer cells;DUSP10-mediated RAD54 B affects the malignant biological behavior of liver cancer cells may be related to activation of MAPK signaling pathway.