Establishment and characterization of a new human ampullary carcinoma cell line,DPC-X1

BACKGROUND An in-depth study of the pathogenesis and biological characteristics of ampullary carcinoma is necessary to identify appropriate treatment strategies. To date, only eight ampullary cancer cell lines have been reported, and a mixed-type ampullary carcinoma cell line has not yet been reported.AIM To establish a stable mixed-type ampullary carcinoma cell line originating from Chinese.METHODS Fresh ampullary cancer tissue samples were used for primary culture and subculture. The cell line was evaluated by cell proliferation assays, clonal formation assays, karyotype analysis, short tandem repeat(STR) analysis and transmission electron microscopy. Drug resistances against oxaliplatin, paclitaxel, gemcitabine and 5-FU were evaluated by cell counting kit-8 assay. Subcutaneous injection 1 × 106 cells to three BALB/c nude mice for xenograft studies. The hematoxylin-eosin staining was used to detect the pathological status of the cell line. The expression of biomarkers cytokeratin 7(CK7), cytokeratin 20(CK20), cytokeratin low molecular weight(CKL), Ki67 and carcinoembryonic antigen(CEA) were determined by immunocytochemistry assay.RESULTS DPC-X1 was continuously cultivated for over a year and stably passaged for more than 80 generations;its population doubling time was 48 h. STR analysis demonstrated that the characteristics of DPC-X1 were highly consistent with those of the patient’s primary tumor. Furthermore, karyotype analysis revealed its abnormal sub-tetraploid karyotype. DPC-X1 could efficiently form organoids in suspension culture. Under the transmission electron microscope, microvilli and pseudopods were observed on the cell surface, and desmosomes were visible between the cells. DPC-X1 cells inoculated into BALB/C nude mice quickly formed transplanted tumors, with a tumor formation rate of 100%. Their pathological characteristics were similar to those of the primary tumor. Moreover, DPC-X1 was sensitive to oxaliplatin and paclitaxel and resistant to gemcitabine and 5-FU. Immunohistochemistry showed that the DPC-X1 cells were strongly positive for CK7, CK20, and CKL;the Ki67 was 50%, and CEA was focally expressed.CONCLUSION Here, we have constructed a mixed-type ampullary carcinoma cell line that can be used as an effective model for studying the pathogenesis of ampullary carcinoma and drug development.

Analysis of textbook outcomes for ampullary carcinoma patients following pancreaticoduodenectomy

BACKGROUND Textbook outcomes(TOs)have been used to assess the quality of surgical treatment for many digestive tumours but not ampullary carcinoma(AC).AIM To discuss the factors associated with achieving a TO and further explore the prognostic value of a TO for AC patients undergoing curative pancreaticoduodenectomy(PD).METHODS Patients who underwent PD at the China National Cancer Center between 1998 and 2020 were identified.A TO was defined by R0 resection,examination of≥12 Lymph nodes,no prolonged hospitalization,no intensive care unit treatment,no postoperative complications,and no 30-day readmission or mortality.Cox regression analysis was used to identify the prognostic value of a TO for overall survival(OS)and recurrence-free survival(RFS).Logistic regression was used to identify predictors of a TO.The rate of a TO and of each indicator were compared in patients who underwent surgery before and after 2010.RESULTS Ultimately,only 24.3%of 272 AC patients achieved a TO.A TO was independently associated with improved OS[hazard ratio(HR):0.443,95%confidence interval(95%CI):0.276-0.711,P=0.001]and RFS(HR:0.379,95%CI:0.228-0.629,P<0.001)in the Cox regression analysis.Factors independently associated with a TO included a year of surgery between 2010 and 2020(OR:4.549,95%CI:2.064-10.028,P<0.001)and N1 stage disease(OR:2.251,95%CI:1.023-4.954,P=0.044).In addition,the TO rate was significantly higher in patients who underwent surgery after 2010(P<0.001)than in those who underwent surgery before 2010.CONCLUSION Only approximately a quarter(24.3%)of AC patients achieved a TO following PD.A TO was independently related to favourable oncological outcomes in AC and should be considered as an outcome measure for the quality of surgery.Further multicentre research is warranted to better elucidate its impact.

Impact of perioperative blood transfusion on oncological outcomes in ampullary carcinoma patients underwent pancreaticoduodenectomy

BACKGROUND The effect of perioperative blood transfusion(PBT)on the prognosis of ampullary carcinoma(AC)is still debated.AIM To explore the impact of PBT on short-term safety and long-term survival in AC patients who underwent pancreaticoduodenectomy.METHODS A total of 257 patients with AC who underwent pancreaticoduodenectomy between 1998 and 2020 in the Cancer Hospital,Chinese Academy of Medical Sciences,were retrospectively analyzed.We used Cox proportional hazard regression to identify prognostic factors of overall survival(OS)and recurrencefree survival(RFS)and the Kaplan-Meier method to analyze survival information.RESULTS A total of 144(56%)of 257 patients received PBT.The PBT group and nonperioperative blood transfusion group showed no significant differences in demographics.Patients who received transfusion had a comparable incidence of postoperative complications with patients who did not.Univariable and multivariable Cox proportional hazard regression analyses indicated that transfusion was not an independent predictor of OS or RFS.We performed Kaplan-Meier analysis according to subgroups of T stage,and subgroup analysis indicated that PBT might be associated with worse OS(P<0.05)but not RFS in AC of stage T1.CONCLUSION We found that PBT might be associated with decreased OS in early AC,but more validation is needed.The reasonable use of transfusion might be helpful to improve OS.

Multiple imaging techniques in the diagnosis of ampullary carcinoma

BACKGROUND: Ampullary carcinoma is a neoplasia with a good prognosis compared to pancreatic cancer. But it is difficult to early diagnose because it lacks clear clinical symptoms. This study aimed to evaluate the efficacy of abdominal ultrasonography (US), enhanced computed tomography (CT), magnetic resonance cholangiopancreatography (MRCP) and endoscopic retrograde cholangiopancreatography (ERCP) in detecting ampullary carcinoma. METHODS: Forty-one patients with ampullary carcinoma who had been confirmed pathologically among the inpatients at the First Affiliated Hospital of Zhejiang University School of Medicine from February 2003 to March 2007 were analyzed retrospectively. The accuracy of US, CT, MRCP and ERCP were compared in the diagnosis of ampullary carcinoma. RESULTS: The accurate rate for detection of ampullary carcinoma with US was 26.83%. The accuracy of CT and ERCP in detection of ampullary tumors was 84.62% and 100%, respectively, which were significantly higher than that of US (P<0.05). The accuracy of MRCP in detection of ampullary tumors was similar to that of US in spite of visualization of obstruction and dilatation of the pancreaticobiliary duct with MRCP. CONCLUSIONS: Because of the obscure and late onset of symptoms, ampullary carcinoma is difficult to diagnose early. Multiple imaging techniques should be carried out appropriately in order to early diagnose the disease and improve the prognosis.

Proposal of a new T-stage classification system for ampullary carcinoma based on Surveillance,Epidemiology and End Result(SEER)database

Background:Tumor size is still considered a useful prognostic factor in currently available tumor-nodemetastasis(TNM)classification staging systems for most solid tumors,but the significance of tumor size on the prognosis of ampullary carcinoma remains controversial.The aim of the current study was to propose a new T-stage classification system for ampullary carcinoma to address the impact of tumor size on the prognostic outcome.Methods:Using the Surveillance,Epidemiology,and End Results(SEER)database,we identified 1080 patients with ampullary carcinoma who underwent radical surgical resection between 2004 and 2015.Based on the results obtained from analysis of various clinicopathologic factors,a new T-stage classification system was proposed.Results:Among the 1080 patients,618 were men and 462 were women,with a median tumor size of 2.3(range 0.1–12)cm.Using the 7 th edition of the American Joint Committee on Cancer(AJCC)staging manual,we noticed significant differences in overall survival(OS)between T2 vs.T3 tumors(P<0.001)and T3 vs.T4 tumors(P=0.002),but failed to observe significant differences between T1 vs.T2 tumors(P=0.498)in our pair-wise comparison.Using the newly developed T-stage classification system,we were able to differentiate significant differences in OS between T1 vs.T2 tumors(P=0.032),T2 vs.T3 tumors(P<0.001)and T3 vs.T4 tumor(P=0.003)in all pair-wise comparisons.The c-index of the new staging system was 0.653(95%CI:0.629–0.677),showing a better discriminatory power than the 0.636 of the 7 th AJCC staging system(95%CI:0.612–0.660).Conclusions:The new T-stage classification system described herein can better differentiate prognostic outcomes after radical resection in patients with ampullary carcinoma by incorporating tumor size and depth of tumor infiltration.

Preoperative prognostic nutritional index predicts long-term outcomes of patients with ampullary adenocarcinoma after curative pancreatoduodenectomy

BACKGROUND The prognostic nutritional index(PNI),a marker of immune-nutrition balance,has predictive value for the survival and prognosis of patients with various cancers.AIM To explore the clinical significance of the preoperative PNI on the prognosis of ampullary adenocarcinoma(AC)patients who underwent curative pancreaticoduodenectomy.METHODS The data concerning 233 patients diagnosed with ACs were extracted and analyzed at our institution from January 1998 to December 2020.All patients were categorized into low and high PNI groups based on the cutoff value determined by receiver operating characteristic curve analysis.We compared disease-free survival(DFS)and overall survival(OS)between these groups and assessed prognostic factors through univariate and multivariate analyses.RESULTS The optimal cutoff value for the PNI was established at 45.3.Patients with a PNI≥45.3 were categorized into the PNI-high group,while those with a PNI<45.3 were assigned to the PNI-low group.Patients within the PNI-low group tended to be of advanced age and exhibited higher levels of aspartate transaminase and total bilirubin and a lower creatinine level than were those in the PNI-high group.The 5-year OS rates for patients with a PNI≥45.3 and a PNI<45.3 were 61.8%and 43.4%,respectively,while the 5-year DFS rates were 53.5%and 38.3%,respectively.Patients in the PNI-low group had shorter OS(P=0.006)and DFS(P=0.012).In addition,multivariate analysis revealed that the PNI,pathological T stage and pathological N stage were found to be independent prognostic factors for both OS and DFS.CONCLUSION The PNI is a straightforward and valuable marker for predicting long-term survival after pancreatoduodenectomy.The PNI should be incorporated into the standard assessment of patients with AC.

Prognostic significance of preoperative Naples prognostic score on short-and long-term outcomes after pancreatoduodenectomy for ampullary carcinoma

Background:The Naples prognostic score(NPS)is an effective and objective tool to assess the immune-nutritional status of patients with malignant tumors.The aim of this study was to investigate the clinical significance of preoperative NPS on short-and long-term outcomes after pancreatoduodenectomy(PD)for ampullary carcinoma.Methods:We retrospectively analyzed 404 consecutive patients with ampullary carcinoma who underwent PD between January 2012 and June 2018.Preoperative NPS was calculated from serum albumin and total cholesterol concentrations,and the neutrophil-lymphocyte ratio and lymphocyte-monocyte ratio(LMR).Patients were then divided into three groups according to their NPS.Clinicopathological variables,postoperative outcomes,and survival data were compared between the three groups.Univariate and multivariate Cox analysis of overall survival(OS)and recurrence-free survival(RFS)were also conducted,and time-dependent receiver operating characteristic(ROC)curves were created to evaluate the discriminatory ability of the prognostic scoring systems.Results:Patients with higher NPS had worse prognosis,and significant OS difference(group 0 vs.1,P=0.02;group 1 vs.2,P<0.001;group 0 vs.2,P<0.001)and RFS difference(group 0 vs.1,P=0.088;group 1 vs.2,P<0.001;group 0 vs.2,P<0.001).Multivariate analysis revealed that NPS was an independent significant predictor of OS(grade 2 vs.grade 1 or 0,hazard ratio:3.067;P<0.001)and RFS(grade 2 vs.grade 1 or 0,hazard ratio:2.732;P<0.001).The time-dependent receiver operating curve analysis showed that NPS had better prognostic performance for OS and RFS than other prognostic models.Additionally,significant differences in the incidence of postoperative morbidity were observed between the three groups,and the NPS was an independent risk factor of overall postoperative complications(grade 2 vs.grade 1 or 0,odds ratio:1.692;P=0.02).Conclusions:The NPS was an independent predictor of overall-and RFS in patients undergoing PD for ampullary carcinoma,and was independently associated with the incidence of postoperative complications.