Detection Of Multidrug Resistance Protein Expression And Mutation Sites In Gastric-type Cervical Adenocarcinoma

BackgroundJapanese scholars study found that the Gastric-Type endocervical adenocarcinoma(GAC)accounted for about 25%of cervical adenocarcinoma,with aggressive strong,easy early lymph node metastasis,and is not sensitive to chemotherapy drugs,poor prognostic features,but at the moment,the study is less,its etiology is unclear,the literature reported that most GAC is found with uterus,ovaries,omentum and celiac metastasis.Multidrug resistance(MDR)refers to the drug resistance of tumor cells to one chemotherapy drug during chemotherapy,and also to other chemotherapy drugs of the same or different types.Mdrl-encoded P-gp is a member of the ATP binding cassette(ABC)family.It is a transmembrane transport protein embedded in the serous membrane of cells and contains a highly conserved atp-binding site with atpase activity.Type of this study was to explore the GAC with UEAC multi-resistant P-gp protein expression,protein research P-gp influence cervical adenocarcinoma drug resistance molecular mechanism,and through the exons sequencing technology to investigate the special mutation GAC,looking for specific diagnostic markers for GAC and individualized therapeutic targets.Chapter ? Immunophenotype and expression of multidrug resistance protein P-gp in Gastric-Type endocervical adenocarcinomaMethod1.The pathological data of new cases of cervical cancer diagnosed by pathology in Qilu Hospital of Shandong University from January 2005 to December 2017 were analyzed by regression analysis.2.Collection of cervical adenocarcinoma cases and wax blocks from 2009 to 2017 in Qilu Hospital of Shandong University,including 1 14 cases of cervical adenocarcinoma,to improve the relevant clinicopathological parameters.3.The expression of P16,MUC6,MUC5AC,CK7 and other proteins in cervical adenocarcinoma tissues were detected by immunohistochemical method,and HPV in cervical adenocarcinoma tissues was detected by PCR.Combined with HE staining of cervical adenocarcinoma tissues,GAC was screened.4.According to the results of immunohistochemistry,HPV detection and HE staining,GAC and UEAC were screened,and the relationship between clinicopathological parameters and prognosis of the two types of cervical adenocarcinoma was compared.5.The expression of P-gp protein in GAC and UEAC was detected by immunohistochemical method,and the relationship between P-gp protein and clinicopathological parameters and prognosis of GAC was analyzed.6.Immunohistochemistry was used to detect the expression of ?-arrestin 2 protein in GAC and UEAC,and to analyze the relationship between P-gp protein and ?-arrestin 2 protein in cervical adenocarcinoma.Result1.From 2005 to 2017,cervical squamous cell carcinoma was the main pathological type of cervical cancer,accounting for the majority,while cervical adenocarcinoma accounted for a minority.In 2549 cases of cervical cancer,2037 cases were cervical squamous cell carcinoma(79.91%)and 512 cases were cervical adenocarcinoma(20.09%).From the composition ratio of cervical adenocarcinoma from 2005 to 2017,the incidence of cervical adenocarcinoma is gradually increasing.2.A total of 2549 cases of cervical cancer from 2005 to 2017 were studied.The age distribution was normal.The youngest age was 17 years old and the oldest was 93 years old.The average age was(49.56 ± 11.34)years old.The median age of onset was 49 years old.The peak age of cervical cancer is 41-50 years old,while the proportion of patients under 30 years old is relatively small.3.Among the 2549 cases of cervical cancer in this study,cervical squamous cell carcinoma had the highest proportion in the age group of 41-50 years old(32.99%),while cervical adenocarcinoma had the highest proportion in the age group of 41-50 years old(34.38%).After comparing the number of cases of cervical squamous cell carcinoma and cervical adenocarcinoma,the results showed that ?2 = 4.388,p=0.036,the difference was statistically significant.This indicates that the age of cervical adenocarcinoma is younger than that of cervical SCC.4.Comparing the clinicopathological features and prognosis of 90 cases of UEAC with 24 cases of GAC,it was found that the differentiation degree of GAC was higher than that of UEAC(p<0.001),GAC was more likely to invade muscle tissue(p=0.008),and GAC was more prone to early lymph node metastasis than that of UEAC(p=0.008).The prognosis of patients with GAC was poor(p=0.0126).There was no significant difference in age,size of tumors between UEAC and GAC(p>0.05).5.Immunohistochemical results showed that the positive rate of P-gp protein in GAC was higher than that in UEAC,and most of them were highly expressed.The expression of P-gp protein in GAC was mostly strong positive and high expression,while that in UEAC was weak positive and low expression(p<0.001,the difference was statistically significant).6.This study analyzed the relationship between the expression of P-gp protein and clinicopathological features in GAC.Chi-square test showed that the expression of P-gp protein was correlated with the degree of differentiation(p=0.022).The expression of P-gp protein was not correlated with age,depth of muscle invasion,tumor size and lymph node metastasis.By Kaplan-Meier analysis,it was found that the expression of P-gp protein had no significant difference in the overall survival of GAC and UEAC(p>0.05).7.Among the 90 cases of UEAC,the co-expression of ?-arrestin 2 and p-gp protein was found in 17 cases,accounting for 18.89%(17/90).The expression of ?-arrestin 2 and p-gp protein was negative in 40 cases,accounting for 44.44%(40/90).After statistical analysis,it was found that ?-arrestin 2 was positively correlated with p-gp in UEAC,and Spearman correlation was 0.351,which was statistically significant(p<0.01).Conclusion1.GAC are generally well differentiated and difficult to differentiate.MUC6,P16 immunostaining and high-risk HPV detection can help to differentiate GAC.2.The differentiation degree of GAC is higher than that of UEAC,which is generally well-differentiated.GAC is more prone to early lymph node metastasis and high invasion than UEAC,and the overall survival of patients with GAC is lower than that of UEAC.3.The high expression of P-gp protein in GAC is higher than that in UEAC,which may indicate that the insensitivity of patients with GAC to chemotherapy drugs is related to the high expression of P-gp protein,providing experimental basis for the individualized treatment of cervical adenocarcinoma.4.P-gp protein was positively correlated with the expression of ?-arrestin 2 in UEAC.Chapter ? Application of total exon sequencing technologyMethodCollect 5 cases GAC,4 cases of UEAC,4 cases of normal cervical tissue for all exons sequencing and analysis of mutant gene biology information,according to the mutation frequency,GO,KEGG enrichment analysis factors such as screen mutations in the genes of interest,in DNA level analysis GAC proliferation behavior and its particularity metastasis related genes,in order to explore the pathogenesis of GAC and GAC specific diagnostic markers.ResultThrough the comparison between groups,GO and KEGG enrichment analysis reports and related literature,found in GAC,PRAMEF4 IP6K2,MUC4,GDI2,RGS12,CD81,PSMA1,MADD,DCTN2,RAE1 gene mutations,due to the genes involved in the body of invasion and metastasis of tumor,epithelial-interstitial transformation,and the cell growth and apoptosis signaling pathways of transduction process,so we guess these gene locus mutation and occurrence and development is closely related to the GAC,And it is closely related to the special biological behavior of GAC prone to early invasion and lymph node metastasis.