Transcription Factor FOXM1 Promotes Proliferation And Invasion In Hepatocellular Carcinoma By Regulating Of TPX2

Part one Bioinformatics analysis of hub genes related to the progression and prognosis of hepatocellular carcinomaObjective:Hepatocellular carcinoma(HCC)is one of the major causes of cancer-related mortality in the world and one of the most prevalent malignancies in China.HCC is the main type of primary liver cancer,which accounts for 75%-85% of all cases.Despite the continuous development of novel treatment strategies,the 5-year survival rate of HCC is still very low,especially in advanced HCC.Therefore,it is important to understand the precise mechanism involved into the development and progression from liver cirrhosis to HCC.Material and Method:(1)The microarray datasets of HCC in GEO database were used to find the significantly differentially expressed genes between liver cirrhosis and HCC.PPI network was constructed to find the hub genes related to the progression of hepatocellular carcinoma.(2)Oncomine,UALCAN and HPA databases were used to verify the expression levels of key genes.(3)Clinical data of hub genes were analyzed using clinical sample datasets of HCC in TCGA database.First,Kaplan-Meier method was used for survival analysis,and then Cox model was established.Finally,the genes with significant differences between the two types of survival analysis were intersected to identify the genes closely related to survival in HCC.(4)Oncomine,c Bio Portal and GEPIA online databases and UCSC Genome Browser were used to predict co-expression genes and upstream transcription factors of TPX2.Result:(1)The 4 gene chips in the GEO database were used to analyze the differentially expressed genes,and 360 differentially expressed genes were obtained.10 hub genes were screened by PPI network.(2)The expression of hub genes in ONCOMINE,UALCAN and HPA databases were all higher than that in normal liver tissues.(3)Based on the clinical data of HCC in TCGA,Kaplan-Meier survival analysis and Cox model revealed that there were 3 genes associated prognosis of HCC.Among them,TPX2 was the most significant.(4)Co-expression analysis using ONCOMINE,c Bio Portal and GEPIA databases showed that the expression level of TPX2 was highly and positively correlated with FOXM1.UCSC Genome Browser analysis revealed that FOXM1 was an upstream transcription factor involved in the regulation of TPX2.Part tow The effects of FOXM1 on the biological functions of HCC cellsObjective:Forkhead box(FOX)protein family is a group of highly conserved transcription factors,which plays an important role in controlling cell cycle,epithelial differentiation,metabolism and immune system regulation and many other biological processes.FOXM1 is an important member of the FOX family.Abnormal expression of FOXM1 can activate cyclin of HCC and promote cell proliferation,which is closely related to the occurrence and development of HCC.However,the specific mechanism of FOXM1 in the occurrence and development of HCC remains unclear.Targeting protein for xklp2(TPX2)is a proliferation-related protein,which is involved in the regulation of cell mitosis and cell cycle.This study will focus on investigating the effects of FOXM1 and its possible target gene TPX2 on the biological behaviors of HCC cells.It is hoped to to provide new research basis for gene targeted therapy of HCC.Material and Method:(1)The protein and gene expression levels of FOXM1 and TPX2 were analyzed by Western blot and RT-q PCR using normal liver cell line L02 and HCC cell lines Hep G2,Huh7 and SMMC-7721.(2)siRNA-FOXM1 was transfected into HCC cell lines to silence FOXM1,the protein expression levels of FOXM1 and TPX2 were analyzed by Western blot,CCK-8,colony formation,wound healing and transwell chamber,flow cytometry were used to determine the effects of siRNA-FOXM1 on cell proliferation,migration,invasion and apoptosis level of HCC cells.Result:(1)The gene and protein expression levels of FOXM1 and TPX2 in liver cancer cell lines Hep G2,Huh7 and SMMC-7721 were higher than those in normal liver cell line L02.(2)Compared with control group and siRNA-NC group,siRNA-FOXM1 reduced the protein expression of FOXM1 and TPX2 in Hep G2 cells.Moreover,siRNA-FOXM1 reduced proliferation,migration and invasion ability,and increased cell apoptosis of Hep G2 cells.Conclusion:1.AURKA,CENPF,HMMR,TOP2 A,Asp M,CCNA2,UBE2 C,RRM2,CDKN3,and TPX2 play important roles in the progression of liver cirrhosis to HCC.2.TPX2 is the most closely related to the prognosis of HCC.3.The upstream transcription factor of TPX2 is predicted to be FOXM1 by bioinformatics method.4.FOXM1 and TPX2 are significantly overexpressed in HCC cell lines.5.The expression level of TPX2 is related to FOXM1,and TPX2 may be the target gene of FOXM1.6.The down-regulated expression of FOXM1 significantly affects the proliferation,invasion,migration and apoptosis levels of HCC cells,suggesting that FOXM1 as an oncogene in HCC.