The Role Of Inflammatory Factors In The Prognosis Of Gallbladder Cancer And The Role Of Non-coding RNA In The Pathogenesis Of Intrahepatic Cholangiocarcinoma Based On CeRNA Regulatory Network Analysis

AIM To investigate the prognostic role of preoperative fibrinogen-toalbumin ratio（FAR）on patients with gallbladder cancer（GBC）in this study.METHODS One hundred and fifty-four GBC patients were retrospectively analyzed,who received surgery in our hospital from March 2005 to December 2017.Receiver operating characteristic curve（ROC curve）was used to determine the optimal cut-offs for these biomarkers including preoperative fibrinogen to albumin ratio.Univariate Kaplan-Meier and multivariate Cox regression analyses were used to perform prognostic analysis of patients with GBC.RESULTS ROC curve revealed that the optimal cut-off value for preoperative FAR was 0.08.preoperative FAR was significantly correlated with age（P=0.045）,jaundice（P<0.001）,differentiation（P=0.002）,resection margin status（P<0.001）,T stage（P<0.001）,TNM stage（P<0.001）,and CA199（P<0.001）as well as albumin levels（P<0.001）.Multivariate analysis indicated that the resection margin status[hazard ratio（HR）:2.343,95%confidence interval（CI）:1.532-3.581,P<0.001],TNM stage（P=0.035）,preoperative albumin level（HR=0.595,95%CI:0.385-0.921,P=0.020）and FAR（HR:2.813,95%CI:1.765-4.484,P<0.001）were independent prognostic factors in GBC patients.CONCLUSION An elevated preoperative FAR was significantly correlated with unfavorable overall survival（OS）in GBC patients,while an elevated preoperative albumin level was a protective prognostic factor for patients with GBC.The preoperative FAR could be used to predict the prognosis of GBC patients,which was easily accessible,costeffective and noninvasive.AIM To investigate the prognostic value of the combination of preoperative plasma fibrinogen and CA199 in patients with gallbladder carcinoma（GBC）.METHODS The clinicopathological data of 154 GBC patients were retrospectively reviewed after surgery.A receiver operating characteristic（ROC）curve was plotted to verify the optimum cut-off values for plasma fibrinogen and CA199.Univariate and multivariate survival analyses were performed to identify the factors associated with GBC prognosis,based on the relative value of hazard ratios（HRs）calculated via multivariate survival analyses,patients with elevated plasma fibrinogen and CA199 levels were allocated a score of 2.1;those with an elevated plasma fibrinogen level only were allocated a score of 1,those with an elevated CA199 level only were allocated a score of 1.1,and those with neither of these abnormalities were allocated a score of 0.RESULTS ROC curve analysis showed that the optimum cut-off values for preoperative plasma fibrinogen and CA199 were 3.47 g/L and 25.45 U/mL,respectively.Multivariate analysis indicated that elevated preoperative plasma fibrinogen and CA199 levels were significantly correlated with worse overall survival（OS）（HR = 1.711,95% confidence interval（CI）: 1.114-2.627,P = 0.014? and HR = 1.842,95%CI: l.lll-3.056? P = 0.018）.When we combined these two parameters,the area under the ROC curve increased from0.735（for preoperative plasma fibrinogen only）and 0.729（for preoperative CA199 oiily）to 0.765.When this combined variable was added to the multivariate analysis,the combination of plasma fibrinogen and CA199（P < 0.001）,resection margin（P < 0.001）and TNM stage（P = 0.010）were independent prognostic factors for GBC patients.CONCLUSION The combination of plasma fibrinogen and CA199 may serve as a more efficient independent prognostic biomarker for postoperative GbC patients than either parameter alone.Background: The purpose of the study is to discuss the prognostic value of lymphocyte-to-monocyte ratio（LMR）in patients with gallbladder carcinoma（GBC）.Methods: In this study,we made a collection of 154 consecutive GBC patients who were hospitalized in our hospital for surgery from 2005 to 2017 retrospectively.Divide the lymphocyte count by the monocyte count in preoperative peripheral blood so as to calculate the LMR of preoperative blood samples.Employ receiver operating characteristic（ROC）curve in order to obtain the optimal cut-off value of LMR in over^l survival（OS）determination.Utilize Kaplan-Meier method so as to assess OS and log-rank test compare difference in survival in different patitent groups.Conduct Univariate and multivariate Cox regression analyses to make detection of independent prognostic indicators for the GBC patients.Results: The ROC curve indicates that the optimal cut-off point for the LMR in preoperative peripheral blood reached 4.76.Compared to those with an LMR >4.76（hazard ratio（HR）: 0.399,95% confidence interval（Cl）: 0.265-0.602,P<0.001）,GBC patients who are <60 years old with an LMR <4.76 has evidently worse OS;nevertiieless,the prognostic value of LMR has no detection among the whole study cohort or in patients > 60 years old（both P>0.05）.Compared to those with an LMR >4.21（HR: 1.830,95% Cl: 1.129-2.967,P=0.014）? we have observed obviously poorer OS in patients >60years with an LMR <4.21.Multivariate Cox regression analysis showed that biomarker LMR which yielded higher and lower LMR cutoff values based on age stratification was independent risk factor for OS in GBC patients of different age groups（HR: 0.272,95%Cl: 0.105-0.704,P=0.007;HR: 0.544,95% Cl: 0.330-0.895,P=0.017）.Conclusions: LMR was an independent prognostic indicator of GBC patients,of which the optimal cut-off value has certain dependence on age.Background: Competitive endogenous RNA（ceRNA）is a novel gene expression regulation model,and it is believed that long non-coding RNA（IncRNA）and genes can competitively bind microRNAs（microRNAs,miRNA）thus plays an important regulatory role in the pathogenesis of cancer.However,the competitive mechanism of ceRNA in intrahepatic cholangiocai*cinoma is not fully understood.Materials and Methods: The IncRNA,miRNA and mRNA dataset expression data of ICC were obtained through retrieving the NCBI GEO database.The differential expression of the above datasets was analyzed and the intersections were taken,so as to obtain the differentially expressed IncRNAs,miRNAs and mRNAs of ICC.In addition,the ICC-related ceRNA regulating network（ceRNET）was constructed through co-expression analysis of the differentially expressed IncRNAs and mRNAs,miRNA target gene prediction analysis,and ceRNA relationship integration;besides,the differentially expressed mRNAs in the network were performed functional enrichment analysis.Subsequently,the ICC-related core regulatory pathway to be verified was obtained according to the network node ranking of the differentially expressed RNAs and consistency of the expression trend of each GEO dataset,based on the relationship of log-fold change（logFC）with ceRNA as well as the previous research results in other tumors.Aftei*wards,such regulatory pathway was experimentally verified using 10 pairs of fresh carcinoma and para-carcinoma tissues from ICC patients,10 peripheral plasma specimens from ICC patients and matched peripheral plasma specimens from 10 healthy subjects,as well as 88 paraffin sections from ICC patients through real-time quantitative polymerase chain reaction（PCR）,Western blotting and immunohistochemical staining.Results: The constructed ICC-related ceRNET had contained 340 lncRNA-miRNAmRNA regulatory relationships.GO analysis of the 44 differentially expressed mRNAs in ceRNET suggested that,they were mainly emiched in the biological processes such as"negative regulation of epithelial cell proliferation” and “positive regulation of the activated T-lymphocyte proliferation”;in addition,KEGG analysis indicated that they were mainly enriched in the “complement and coagulation cascade” pathway.RP11-328K4.1-hsa-miR-27a-3p-PROS1 was the ICC-related core regulatory pathway in ceRNET determined in this study.Molecular biological experiment verified that,compared with normal control,lncRNA RP11-328K4.1 expression in cancer tissues and peripheral plasma of ICC patients was down_regulated,and the differences were statistically significant（p=0.000007 and 0.036093）.hsa-miR-27a-3p expression in cancer tissues and peripheral plasma of ICC patients was up-regulated5 and the differences were of statistical significance（p= 0.00016 and 0.049420345）.Meanwhile,low mRNA PROS1 expression could be detected in cancer tissues and peripheral plasma of ICC patients,and the difference between cancer and para-carcinoma tissues was statistically significant（p=0.006611）,while tliat between peripheral plasma was not statistically significant（p=0.171259）.The expression of proteins corresponding to mRNA PROS1 was down-regulated relative to that of adjacent normal tissues,whereas,in the peripheral plasma of ICC patients was up-regulated relative to that of normal subjects,but the difference was not statistically significant in either group（p=0.668353048 and0.597799476）;in the ICC paraffin pathological sections,the staining of protein PROS1 in cancer tissues was positive compared with that in para-carcinoma tissues.Conclusions: Constructing ceRNET is an effective approach to investigate the pathogenesis of ICC.The occurrence of ICC is related to the regulation of proliferation of epithelial cells and the activated T lymphocytes,and the complement and coagulation cascade pathway is involved in the pathogenesis of ICC.The up-regulated IncRNA RP11-328K4.1 expression can remove the suppression of the oncogenic miRNA hsa-miR₂7a on mRNA PROS 1 expression through the sponge adsorption effect,thus exerting the role of IncRNA RP11-328K4.1 in ICC as a tumor suppressor gene.IncRNA RP11-328K4.1,miRNA hsa-miR-27a-3p and mRNA PROS1 in the core regulatory pathway are the ICC-related pivot nodes with high connectivity in ceRNET,which are the ideal candidate biomarkers to investigate and discover the diagnostic,prognostic and therapeutic targets for ICC patients in the fixture.