Study On The Expression,Cell Function And Regulatory Mechanism Of IMPA2 In Epithelial Ovarian Cancer

Objective:(1)To identify the expression characteristics of IMPA2 in epithelial ovarian cancer.(2)To elucidate the effect of IMPA2 on the biological function of epithelial ovarian cancer cells.(3)To reveal the regulatory mechanism of IMPA2 on biological function of epithelial ovarian cancer cells.Methods:(1)The expression of IMPA2 and its correlation with clinical features were analyzed based on TCGA,GTEx and CPTACdatabases,and the expression level of IMPA2 gene in four epithelial ovarian cancer cell lines was verified by RT-PCR.(2)IMPA2 lentivirus interference vector was constructed to abtain stable knockdown epithelial ovarian cnacer cell lines,and RT-q PCR and western blot were used to detect the interference efficiency.(3)The influence of IMPA2 on ovarian cancer was detected,such as Celigo cell imaging techniques,MTT assay and clonogenesis assay were used to detect cell proliferation,flow cytometry was used to detect apoptosis,transwell chamber assay was used to detect cell invasion and migration.(4)The tumor formation experiment in nude mice was used to verify the results of in vitro experiments.(5)The databases was used to screen the functions and signaling pathways of IMPA2 enrichment.Western blot was used to detect the expression and phosphorylation levels of signaling pathways related to IMPA2 low expression.Moreover,invasion proteins and epithelial mesenchymal transforming(EMT)proteins were also detected by WB to screen the downstream targets regulated by IMPA2.(6)Stable epithelial ovarian cancer cell lines with low IMPA2 knockdown and VIM overexpression were constructedto detect cell proliferation and metasitasis ablility.Results:(1)IMPA2 was down-expressed in 12 tumors and overexpressed in 14 tumors,and especially IMPA2 is significantly high expression in ovarian cancer..Further,we verified IMPA2 expression in four ovarian cancer cell lines.(2)Pan-cancer analysis showed that high IMPA2 expression was associated with clinical stage Ⅲ,Ⅳ and age <59 years in ovarian cancer patients.The mutation and copy number variation of IMPA2 gene were significantly positively correlated with the occurrence of ovarian cancer.And,the expression of IMPA2 was negatively correlated with differentiated cluster 4(CD4)T cells,and positively correlated with B cells and DC cells(P < 0.05).(3)Lentivirus RNA interference significantly silenced the expression of IMPA2,which significantly inhibited the proliferation and the ability of colony formation,anti-apoptosis,migration and invasion(P < 0.05).(4)Low expression of IMPA2 inhibited the tumorigenic ability of ovarian cancer cells in nude mice by significantly inhibiting the tumor volume and weight(P < 0.05).(5)The enrichment results showed that IMPA2 was involved in nuclear factor κB(NF-κB),Wnt and EMT pathway in ovarian cancer.The results of WB further demonstrated that IMPA2 knockdown may down-regulate the expression of key proteins and their phosphorylated proteins in ERK/P38/MAPK,PI3K/ AKT/ mTOR,Wnt/β-Catenin and NF-κB signaling pathways in SKOV3 cells.Meanwile,IMPA2 regulated the expression of invasive proteins(Twist and MMP9)and EMT protein(Slug,Snail,Twist,CDH1,CDH2,FN1 and VIM)(P < 0.05).(6)IMPA2 regulated the proliferation and metastasisof epithelial ovarian cancer cells by taegeting VIM gene.Conclusion: IMPA2 was highly expressed in ovarian cancer tissues and cell lines.The down-expression of IMPA2 inhibited themalignant biological function of epithelial ovarian cancer cells by targeting VIM.These results suggest that IMPA2 is involved in the regulation of the occurrence and metastasis of epithelial ovarian cancer,providing a new strategic molecular target for the personalized treatment of epithelial ovarian cancer.