| Objective: Endometrial cancer(EC)is one of the three most common malignant tumors of the female reproductive system.The incidence rate is increasing year by year,and the incidence rate of young women is rising sharply.The treatment methods for early endometrial cancer now include surgery,radiotherapy and chemotherapy,and hormone therapy.However,the treatment effect for patients with advanced and recurrent endometrial carcinoma is still very poor.Effective biomarkers or targets are urgently needed to clarify the diagnosis,optimize the treatment of endometrial cancer and evaluate the prognosis.However,new immunotherapy as a potential treatment strategy for EC patients has attracted widespread attention.Therefore,it is very important to explore and develop reliable early diagnostic markers,immune-related targets and prognostic markers for the individualized treatment of EC.CDKN2B antisense RNA1(CDKN2B-AS1)participates in a variety of regulatory roles,including the cell cycle of tumors.As a carcinogenic molecule,it is related to several cancers.The increased level of CDKN2B-AS1 m RNA may accelerate the proliferation of tumor cells and inhibit the apoptosis of tumor cells,thus playing a carcinogenic role in various tumors.Indoleamine-2,3 dioxygenase 1(IDO1)is one of the three enzymes that catalyze the first rate-limiting step of tryptophan oxidation metabolism.Tryptophan is an essential amino acid for T cell proliferation and differentiation,and may participate in immune escape through local Trp depletion and the formation of deadly Trp catabolism products.More and more evidence shows that IDO1 is overexpressed in most malignant tumors.Gene expression data obtained from TCGA database and immune marker samples show that cervical cancer has the highest expression of IDO1,followed by endometrial cancer,bladder cancer cancer,kidney cancer and lung cancer.Studies have shown that overexpression of IDO1 can promote the escape of malignant tumors from immune monitoring.Therefore,we explore that CDKN2B-AS1 can affect the malignant biological behavior of endometrial cancer cell lines by regulating the expression of IDO1.Method: In this study,we analyzed the TCGA database of endometrial cancer and found lnc RNA,PRRT3-AS1,LINC01503,CDKN2B-AS1,LINC01629,LINC01833,LINC01936 related to the immune microenvironment,and analyzed the correlation between their expression and the poor prognosis of patients.The correlation between its expression and immune cell infiltration,common immune checkpoints,MHC molecule,chemokine and chemokine receptor was analyzed.We verified the expression level of PRRT3-AS1,LINC01503,CDKN2B-AS1,LINC01629,LINC01833 and LINC01936 in EC and normal endometrial tissues through Realtime-PCR.Next,we evaluated the effect of CDKN2B-AS1 knockout on the malignant biological behavior of EC cell lines by transfecting Ishikawa and HEC1 A cells with sh-CDKN2B-AS1 chronic virus vector and corresponding NC.A stable transfection cell line silencing CDKN2B-AS1 was established in endometrial cancer cell lines Ishikawa and HEC-1A.After confirming transfection efficiency using quantitative real-time PCR,Ed U was used to detect the proliferation ability of cells and Transwell technology was used to detect the invasion ability of cells,next,we studied the function of CDKN2B-AS1 in tumor cell and in vivo model.Silence or overexpression of IDO1 in endometrial cancer cells was analyzed to analyze its effect on the proliferation and invasion of Ishikawa and HEC-1A cells.The proliferation and invasion ability of cells were detected by Ed U and Transwell,respectively.To verify whether CDKN2B-AS1 controls the regulation of IDO1-mediated malignant behavior of endometrial cancer cells,we conducted co-transfection in Ishikawa and HEC-1A cells to study the exact contribution of CDKN2B-AS1-IDO1 axis to Ishikawa and HEC-1A cells.Results: We jointly analyzed from TCGA database and Imm Lnc official website,and screened the differentially expressed immune-related lnc RNAs.Univariate Cox and multivariate Cox regression finally identified six immune-related lnc RNAs as prognostic markers: PRRT3-AS1,LINC01503,LINC01936,CDKN2B-AS1,LINC01629,LINC01833.Through multivariate Cox regression analysis of age,clinical stage,tumor differentiation and invasion depth,it was confirmed that six immune-related lnc RNAs,PRRT3-AS1,LINC01503,LINC01936,CDKN2B-AS1,LINC01629,LINC01833,were independent prognostic factors.We detected the expression of PRRT3-AS1,LINC01503,LINC01936,CDKN2B-AS1,LINC01629,LINC01833 in EC and normal endometrial tissues by q PCR.The results showed that CDKN2B-AS1 was significantly overexpressed in human endometrial carcinoma,suggesting that CDKN2B-AS1 might play a key role in the development of endometrial carcinoma.A stable cell line silencing CDKN2B-AS1 was established in endometrial cancer cell lines Ishikawa and HEC-1A.Interference with CDKN2B-AS1 can inhibit the proliferation of Ishikawa and HEC-1A cells,and interference with CDKN2B-AS1 can also inhibit its invasive ability.Next,the function of silencing CDKN2B-AS1 in Ishikawa tumor model in vivo found that silencing CDKN2B-AS1 inhibits the growth of nude mice transplanted tumor.It is suggested that CDKN2B-AS1,as a tumor promoting molecule,affects the proliferation and invasion of endometrial cancer cells.Through cell experiments of interference and overexpression of IDO1,the results showed that IDO1,as a oncogenic gene,existed in Ishikawa and HEC-1A cells,and promoted the proliferation and invasion of Ishikawa and HEC-1A cells.When Ishikawa and HEC-1A cells were co-transfected with IDO1(+)and sh-CDKN2B-AS1,IDO1(+)increased the ability of sh-CDKN2B-AS1 to proliferate and invade cells,and the overexpression of IDO1 could reverse the anti-tumor effect of shCDKN2B-AS1.Conclusion: 1.PRRT3-AS1,LINC01503,LINC01936,CDKN2B-AS1,LINC01629,LINC01833 are immune-related lnc RNAs of endometrial carcinoma.2.CDKN2B-AS1 can promote the malignant biological behavior of endometrial cancer cells.3.CDKN2B-AS1 can regulate the malignant biological behavior of endometrial cancer cells by positively regulating the expression of IDO1. |
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