| Objective:This study was to investigate the relationship between clinicopathological features and prognosis of endometrial serous carcinoma(USC)and clear cell carcinoma(UCCC).The relationship between the expression of bispecific phosphatase-1(DUSP1)and the prognosis of endometrial carcinoma was further investigated.Methods:The clinicopathological data of patients with USC and UCCC who underwent surgical treatment in Cancer Hospital of Peking Union Medical College,Chinese Academy of Medical Sciences from January 1,2004 to December 31,2014 were collected.The diagnosis,treatment and prognosis were retrospectively analyzed.Kaplan Meier method and Cox regression analysis were used for univariate and multivariate survival analysis.From January 1,2010 to December 31.2014,the expression of DUSP1 in paraffin sections of USC and UCCC patients was detected by immunohistochemistry,and the relationship between DUSP1 expression level and clinicopathological features and prognosis was analyzed.Results:Research results of USC patients:The study included 71 patients.Only 32 patients(45.1%,32/71)were diagnosed with USC preoperatively and 25 patients(35.2%,25/71)underwent USC standard comprehensive staging surgery.Of the 25 patients,10(40.0%.10/25)patients had up-staged after operation.The 5-year disease free survival[DFS]rate and overall survival[OS]rate for all patients were 76.5%and 80.6%respectively.On univariate analysis,age,range of lymphadenectomy,tumor stage,peritoneal cytology and the depth of myometrial invasion were associated with 5-year DFS rate(P<0.05);range of lymphadenectomy,range of surgical staging,tumor stage,peritoneal cytology and postoperative adjuvant treatment were associated with 5-year OS rate(P<0.05).On multivariate analysis,range of surgical staging(Incomplete surgical staging vs Complete surgical staging.HR=5.18.P<0.05)and tumor stage(?A vs IA,HR=34.99,P<0.01;?B vs ?A,HR=96.50,P<0.01)were independent prognostic factors for 5-year DFS rate;range of lymphadenectomy(no vs PLN+PALN,HR=27J6,P<0.05;PLN vs PLN+PALN,HR=5.98,P<0.05)and peritoneal cytology(positive vs negative,HR=5.47.P<0.05)were independent prognostic factors for OS.Research results of UCCC patients:The study included 34 patients.Only 18 patients(52.9%)were diagnosed with UCCC preoperatively and 8 patients(23.5%)underwent UCCC standard comprehensive staging surgery.The 5-year DFS rate and OS rate for all patients were 79.1%and 81.3%,respectively.On univariate analysis,pre-operative CA125,range of lymphadenectomy,tumor stage and peritoneal cytology were associated with 5-year DFS rate(P<0.05);pre-operative CA125,range of lymphadenectomy,tumor stage,peritoneal cytology and Lymph vascular space invasion were associated with 5-year OS rate(P<0.05).On multivariate analysis,peritoneal cytology was the only independent prognostic factor for DFS(positive vs negative,HR=11.47,P<0.01);tumor stage was the only independent prognostic factor for OS(?B vs IA,HR=25.29,P<0.01).Research results of DUSP1 expression in common specific pathological types of endometrial cancer(USC and UCCC):DUSP1 expression in USC and UCCC is expressed in both cytoplasm and nucleus,and mainly in the cytoplasm.In 31 cases of USC and UCCC,18 cases(58.1%)were strongly positive(+++),10 cases(32.3%)were moderately positive(++),1 case(3.2%)was weakly positive(+),and 2 cases(6.5%)were negative.DUSP1 positive was correlated with LVSI(P<0.05).Kaplan-Meier univariate survival analysis showed that the prognosis of LVSI positive patients was worse than that of LVSI negative patients(P<0.05),the 5-year cumulative DFS rates were 80.8%and 76.9%(P=0.987)in patients with strong and non strong expression of DUSP1,and the 5-year cumulative OS rates were 80.8%and 76.9%(P=0.791),respectively.Conclusions:(1)The preoperative pathological diagnosis of USC is difficult,resulting in incomplete surgical staging and inaccurate staging.Range of surgical staging,adnexal and/or serosa involvement,peritoneal cytology and range of lymphadenectomy are independent prognostic factors.(2)The preoperative pathological diagnosis of UCCC is difficult,resulting in incomplete surgical staging.Peritoneal cytology and tumor stage are independent prognostic factors.(3)DUSP1 expression is closely related to Lvsi status in USC and UCCC,which may affect the prognosis.DUSP1 may be a potential marker and therapeutic target of poor prognosis in patients with USC and UCCC.Objective:This study intends to explore the role of DUSP1 in progesterone treatment of endometrial cancer and its relationship with PR through further research,hoping to find more key molecules in progesterone treatment and improve the effectiveness of conservative treatment.Methods:Real time PCR and Western blot were used to detect the gene expression in endometrial adenocarcinoma cell lines.The biological function changes of endometrioid adenocarcinoma cell lines after DUSP1 knockdown were detected.Transwell assay was used to detect cell migration,CCK8 assay was used to detect cell growth,and Annexin-V-FITC/PI double staining flow cytometry was used to detect cell apoptosis.PR was knocked down to detect DUSP1 expression.Statistical analysis:t Test was used for the Comparison of measurement data between two groups,univariate analysis of variance was used for the comparison among multiple groups,and Tukey's Multiple Comparison Test was used for the pair Comparison within the group.When the variances of the two groups are not uniform,non-parametric tests are used between the groups.The software GraphPad Prism8.0 was used for analysis and drawing.The GEPIA database(website:http://gepia.cancer-pku.cn/index.html)was used to analyze the expression of PR and DUSP1 in normal endometrial tissues and endometrial cancer tissues,and the correlation between PR and DUSP1 was analyzed using this database.The String protein interaction network database was used for protein interaction analysis(https://string-db.org/).P<0.05 was considered statistically significant.Results:(1)Ishikawa cells were treated with MPA at different concentrations for different time.With the increase of MPA concentration and the prolongation of MPA treatment time,the migration ability of Ishikawa cells decreased significantly.Transwell results showed that the migration ability of si-DUSP1-Ishikawa cells was significantly enhanced compared with the negative control si-NC-Ishikawa cells.CCK8 assay showed that the growth of si-DUSP1-ishikawa cells was significantly enhanced after treatment with the same MPA concentration and time compared with the negative control si-NC-Ishikawa cells.Annexin-V-FITC/PI double staining flow cytometry was used to detect the apoptosis rate of si-DUSP1-Ishikawa cells before and after DUSP1 gene knockdown.Compared with si-NC-Ishikawa cells,the apoptosis rate of si-DUSP1-Ishikawa cells was significantly decreased after MPA treatment at different concentrations and time.(2)When si-NC-Ishikawa cells were treated with different concentrations of MPA for different time,the results of PCR and Western blot showed that the expression of DUSP1 increased significantly with the increase of MPA concentration or time.After PR knockdown,compared with si-NC-Ishikawa cells,the expression of DUSP1 in si-PR-Ishikawa cells was significantly decreased after MPA treatment at different concentrations and time.These results indicate that MPA can regulate the expression of DUSP1 through PR.(3)Ishikawa cells were transfected with siRNA containing PR gene and empty control siRNA.The expression of DUSP1 was detected 72 hours after transfection.The results showed that the expression of DUSP1 in si-PR-Ishikawa cells was lower than that in si-NC-Ishikawa cells(P<0.05),indicating that PR could directly regulate the expression of DUSP1.(4)The expression of PR and DUSP1 in normal endometrial tissues and endometrial carcinoma tissues(including 174 cases of endometrial carcinoma and 91 cases of normal endometrial tissues)were analyzed by GEPIA database.Compared with normal endometrial tissue,the expression of PR and DUSP1 in endometrial carcinoma tissue was decreased(P<0.05).The correlation analysis between PR and DUSP1 showed that PR was positively correlated with DUSP1(r=0.13,P<0.05).Protein interaction network analysis showed that the interaction between PR and DUSP1 might be through MAPK1,MAPK3,MAPK8,MAPK14 and HSP90AA1(P<0.05).Conclusions:DUSP1 knockdown in endometrial carcinoma Ishikawa cells leads to progesterone resistance.The expression of DUSP1 is closely related to PR,and progesterone can increase the regulation of PR on DUSP1 expression,thus playing an anti-tumor role.Targeted increase of DUSP1 expression may overcome the impaired efficacy of progesterone therapy resistance due to PR deficiency and become a new and promising antitumor therapy. |
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