| Background:ALKBH1,as a DNA 6mA demethylase,can play an important role in the development and progression of tumors by regulating the level of DNA 6mA.Abnormal expression of ALKBH1 has been reported in various tumors such as liver cancer,gastric cancer,and glioblastoma.At present,the expression of ALKBH1 in pancreatic cancer and the effect of ALKBH 1 on the biological behavior of pancreatic cancer cells were still unclear.The purpose of this study was to clarify the expression of ALKBH 1 in pancreatic cancer,initially research the effect of ALKBH1 on the biological behavior of pancreatic cancer cells and lay the foundation for further exploration of the molecular mechanism of ALKBH 1.Methods:The public database of TCGA and GTEx were used to retrieve the expression of ALKBH1 in pancreatic cancer and normal pancreatic tissues.Real-time PCR was used to detect the mRNA expression level of ALKBH1 in the four pancreatic cancer cells including ASPC-1,PANC-1,BXPC-3 and SW1990 and normal pancreatic cell of CCC-HPE-2.The expression of ALKBH1 protein in cancer tissues and normal tissues,as well as pancreatic cancer cell lines and normal pancreatic cells,were detected by western blot.CRISPR-Cas9 was used to construct the ALKBH1 knockout SW1990 pancreatic cancer cell line.And the CCK-8 cell viability assay,CCK-8 cell proliferation test,cell scratch test,and Transwell invasion test were used to compare the activity,proliferation,migration,and invasion of pancreatic cancer cells with different expression levels of ALKBH 1.Results:The analysis of public databases revealed that the expression level of ALKBH 1 in pancreatic cancer was significantly higher than the normal tissues(P<0.01).Western blot showed that among 7 pairs of tissue samples,the expression level of ALKBH 1 protein in cancer tissue was significantly higher than the normal pancreas tissue(P<0.05).Real-time PCR and Western blot results showed that the expression levels of ALKBH1 mRNA and ALKBH1 protein in ASPC-1,PANC-1,BXPC-3,and SW1990 cells were higher than those of CCC-HPE-2 cell(P<0.05).Flow Cytometry and Western blot results showed that the ALKBH 1 knockout SW1990 pancreatic cancer cell lines:ALKBH1-sgl and ALKBH1-sg2 were successfully constructed.The results of CCK-8 cell viability test,CCK-8 cell proliferation test,cell scratch test,and Transwell invasion test showed that the viability,proliferation,migration and invasion ability of pancreatic cancer cells in ALKBH1 knockout group were significantly reduced(P<0.05).Conclusion:This study revealed that ALKBH1 was upregulated in pancreatic cancer tissues and cell lines;knocking out ALKBH1 can inhibit the viability of pancreatic cancer cells and reduce the proliferation,migration and invasion ability of pancreatic cancer cells.Objective:The survival prediction for patients with resected pancreatic adenocarcinoma by the tumor,nodes,metastasis(TNM)system is limited.Nomogram,a user-friendly predictive model,is widely applied for malignancies.The purpose of this study was to develop and validate a nomogram for patients with resected pancreatic adenocarcinoma.Methods:A total of 368 patients(258 in the training set,110 in the validation set)with resected pancreatic adenocarcinoma were included in the China National Cancer Center from January 2008 to October 2018.Nomogram was established based on the results of the Cox multivariate analysis.Validation was performed by discrimination and calibration;the area under receiver operating characteristics curve(AUC)was employed to assess the accuracy of survival predictions.Results:Multivariate analysis in the training set showed that blood transfusion,T-stage,N-stage,grade,capsule invasion,carbohydrate antigen 199,neutrophil percentage,and adjuvant therapy were independent prognostic factors for OS(P<0.05).A nomogram predicting 1-year,3-year,and 5-year OS rates,with favorable calibration,was established based on the independent prognostic factors.The C-indices of the nomogram were higher than the TNM system both in training and validation sets.A clear risk stratification system based on the nomogram assigned patients into three groups:low risk group(≤168),moderate risk group(168-255).and high risk group(>255).The risk stratification system showed better ability than the TNM system in predicting 1-year,3-year,and 5-year OS rates(the clear risk stratification system,AUC:0.758,0.709 and 0.672 vs the TNM system,AUC:0.614,0.604 and 0.568)(P<0.05).Conclusion:We developed and validated a nomogram for patients with resected pancreatic adenocarcinoma by including additional independent prognostic factors,such as tumor marker,immune index,surgical information,pathological data,and adjuvant therapy.Compared to the 8th edition TNM system,the nomogram developed in our study showed better performance in predicting prognosis.Objective:To investigate the prognostic value of three different staging schemes based on the number of lymph node metastasis(N stage),lymph node ratio(LNR)and log odds of positive lymph nodes(LODDS)in pancreatic cancer patients after RO resection.Methods:The clinical and pathological data of 307 pancreatic cancer patients who underwent R0 resection at the Cancer Hospital of Chinese Academy of Medical Sciences from January 2010 to December 2018 were retrospectively analyzed.Kaplan-Meier and Cox proportional hazards regression models were used to analyze the independent prognostic factors.The area under the ROC curve(AUC)was used to compare the prognostic efficacy of different lymph node staging systems.The scatter plot was used to compare the LODDS and LNR staging systems.Results:The median survival was 24.0 months,1-year,3-year,and 5-year survival rates were 72.0%,35.5%,and 24.0%,respectively.Multivariate analysis showed that CA199,differentiation,T stage,adjuvant therapy,N stage,LNR stage,and LODDS stage were independent prognostic factors for pancreatic cancer patients after R0 resection(P<0.05).The AUC values of the 1-year survival rate for N stage,LNR stage,and LODDS stage were 0.591,0.592,and 0.609.The AUC values of the 3-year survival rate for N stage,LNR stage,and LODDS stage were 0.585,0.588,and 0.593.The AUC values of the 5-year survival rate for N stage,LNR stage,and LODDS stage were 0.554,0.557,and 0.589.The AUC values of the LODDS stage were the highest and the N stage were the lowest,but there was no significant difference among these three systems(P>0.05).For all patients,the LODDS staging system was slightly better than the N and LNR staging systems,but the difference was not statistically significant(P>0.05).For patients with NO stage,the LODDS staging system showed better prediction performance than the N stage and LNR staging systems(P<0.05).Conclusion:N stage,LNR stage,and LODDS stage were all independent prognostic factors for pancreatic cancer patients after R0 resection.For patients without lymph node metastasis,LODDS staging system was superior to N and LNR staging systems in predicting prognosis. |
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