Analysis Of Clinical Features And Prognosis Of Non-HPV-Associated Adenocarcinoma Classified By IECC

BackgroundCervical cancer(CC)is the fourth most common cancer of women in the world.Continuous infection with high-risk HPV is recognized as the main cause of CC.With the high sensitivity of HPV test and the wide application of CC screening,the detection rate of CC has increased significantly in recent years.However,there are a small number of HPV-negative cases have missed,and the pathological type is mostly endocervical adenocarcinoma(ECA).In 2018,International Endocervical Adenocarcinoma Criteria and Classification(IECC)combined etiology and morphology to divide ECA into HPV-associated adenocarcinoma(HPVA)and non-HPV-associated adenocarcinoma(NHPVA).The latter is only in the minority,its etiology is unclear and no targeted treatment plan.Bioinformatics and gene sequencing technology based on big data sharing provides new ideas for the clinical diagnosis and treatment of rare and recurrent cancers.PurposesTo analyze the differences of clinical characteristics and prognosis between NHPVA and HPVA,to explore the differential genes of them by bioinformatics analysis,to verify the differences in expression of the genes and clarify their relationship with the prognosis of NHPVA.Methods1.Clinical data collection and analysisThe database of Qilu Hospital was searched for patients of ECA between January 2010 and December 2020.The cases were reviewed with HE slides after screening and divided into HPVA and NHPVA by IECC.Our study retrospectively compared the clinical and pathological features of the two groups,and analyzed the overall survival(OS)and Disease-free survival(DFS)by Kaplan-Meier curves and Log-Rank test.The multivariate Cox regression model was used to explore risk factors affecting prognosis.2.Bioinformatics analysisData were obtained through Gene Expression Omnibus(GEO)and Cancer Genome Atlas(TCGA)databases,and limma,Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)and survival analysis were performed to obtain the target genes.3.The expression of target genes in NHPVA and HPVA was detected by immunohistochemical staining assay,and clarify their relationship with the prognosis of NHPVA.Results1.Analysis of clinical data of NHPVA and HPVA1.1 Among the 336 enrolled cases of ECA,64 cases(39.05%)were NHPVA and 272 cases were HPVA.No significant differences were observed in the age,pregnancy and parturition times,clinical symptoms and thinprep cytologic test(TCT)(all P>0.05),and the sensitivity of NHPVA was 60.6%by TCT.1.2 The neutrophil-to-lymphocyte ratio(NLR)and the positive rates of CA125 and CA199 were higher in the NHPVA group(all P<0.05).1.3 NHPVA was more significantly associated with FIGO stage(≥stage Ⅱ),poor differentiation,tumor diameter(≥4cm),deep stromal invasion(≥l/2),lymph node metastasis and ovarian and oviduct metastasis(all P<0.05).And the positive rate of P53 was higher,while the positive rate of P16 was lower than that of HPVA(all P<0.05).And the multivariate Logistic regression analysis showed poor differentiation is the independent risk factor for lymph node metastasis(P<0.05).1.4 In the NHPVA group,the 3-year and 5-year OS was 81.2%and 69.5%,respectively.Which were lower than those of the HPVA group,and NHPVA was prone to recurrence(all P<0.05).The multivariate Cox regression model showed that poor differentiation and lymph node metastasis were significantly associated with the OS of NHPVA(all P<0.05).2.Bioinformation analysis to clarify the target genesLimma analysis screened 511 differential genes,including 314 upregulated genes and 197 downregulated genes(all P<0.05).GO and KEGG enrichment analysis showed that the differentially expressed genes were related to cell cycle,epithelial cell proliferation and cell adhesion.The differentially expressed gene HENMT1 and APOBEC3B were finally obtained by univariate and multivariate COX regression analysis(all P<0.05).3.Differential expression of HENMT1 and its relationship with the prognosis of NHPVAImmunohistochemical staining assay was used in 80 cases of ECA(32 cases of NHPVA and 48 cases of HPVA).The high and low expression were divided according to immunohistochemical staining score≥6.Our results showed that the proportion of high expression of HENMT1(43.75%)in NHPVA group were lower than those in HPVA group.In addition,Kaplan-Meier survival analysis showed that the DFS and OS of the cases with low expression of HENMT1 were significantly shorter(all P<0.05).Conclusions1.Some clinical and pathological features of NHPVA and HPVA by IECC were significantly different,and NHPVA was prone to recurrence and worse prognosis.Poor differentiation and lymph node metastasis are the risk factors of poor prognosis.2.TCT is extremely important for the screening of ECA,which can reduce the missed diagnosis rate of NHPVA.3.The low expression of HENMT1 was associated with NHPVA and poor prognosis.