MMP14 is a diagnostic gene of intrahepatic cholangiocarcinoma associated with immune cell infiltration

BACKGROUND Intrahepatic cholangiocarcinoma(ICC)is a malignant tumor of the hepatobiliary system with concealed onset,strong invasiveness and poor prognosis.AIM To explore the disease characteristic genes that may be helpful in the diagnosis of ICC and affect immune cell infiltration.METHODS We downloaded two ICC-related human gene expression profiles from GEO database as the training group(GSE26566 and GSE32958 datasets)for difference analysis,and performed enrichment analysis on differential genes.The least absolute shrinkage and selection operator(LASSO),support vector machinerecursive feature elimination(SVM-RFE)and random forest(RF),three machine learning algorithms,were used to screen the characteristic genes.Double verification was carried out on GSE107943 and The Cancer Genome Atlas,two verification groups.Receiver operating characteristic curve and area under the curve(AUC)were used to evaluate the diagnostic efficacy of genes for ICC.CIBERSORT and ssGSEA algorithms were used to evaluate the effect of characteristic genes on immune infiltration pattern.Human Protein Atlas(HPA)was used to analyze the protein expression level of the target gene.RESULTS A total of 1091 differential genes were obtained in the training group.Enrichment analysis showed that the above genes were mainly enriched in small molecular catabolism,complement and coagulation cascade,bile secretion and other functions and pathways.Twentyfive characteristic genes were screened by LASSO regression,19 by SVM-RFE algorithm,and 30 by RF algorithm.Three algorithms were used in combination to determine the characteristic gene of ICC:MMP14.The verification group confirmed that the genes had a high diagnostic accuracy(AUC values of the training group and the verification group were 0.960,0.999,and 0.977,respectively).Comprehensive analysis of immune infiltration showed that MMP14 could affect the infiltration of monocytes,activated memory CD4 T cells,resting memory CD4 T cells,and other immune cells,and was closely related to the expression of CD200,cytotoxic T-lymphocyteassociated antigen 4,CD14,CD44,and other immune checkpoints.The results of immunohistochemistry in HPA database showed was indeed overexpressed in ICC.CONCLUSION MMP14 can be used as a disease characteristic gene of ICC,and may regulate the distribution of immune-infiltrating cells in the ICC tumor microenvironment,which provides a new method for the determination of ICC diagnostic markers and screening of therapeutic targets.

Any role for transarterial radioembolization in unresectable intrahepatic cholangiocarcinoma in the era of advanced systemic therapies?

Intrahepatic cholangiocarcinoma(iCCA)is recognized as the second most frequently diagnosed liver malignancy,following closely after hepatocellular carcinoma.Its incidence has seen a global upsurge in the past several years.Unfortunately,due to the lack of well-defined risk factors and limited diagnostic tools,iCCA is often diagnosed at an advanced stage,resulting in a poor prognosis.While surgery is the only potentially curative option,it is rarely feasible.Currently,there are ongoing investigations into various treatment approaches for unresectable iCCA,including conventional chemotherapies,targeted therapies,immunotherapies,and locoregional treatments.This study aims to explore the role of transarterial radioembolization(TARE)in the treatment of unresectable iCCA and provide a comprehensive review.The findings suggest that TARE is a safe and effective treatment option for unresectable iCCA,with a median overall survival(OS)of 14.9 months in the study cohort.Studies on TARE for unresectable iCCA,both as a first-line treatment(as a neo-adjuvant down-staging strategy)and as adjuvant therapy,have reported varying median response rates(ranging from 34%to 86%)and median OS(12-16 mo).These differences can be attributed to the heterogeneity of the patient population and the limited number of participants in the studies.Most studies have identified tumor burden,portal vein involvement,and the patient’s performance status as key prognostic factors.Furthermore,a phase 2 trial evaluated the combination of TARE and chemotherapy(cisplatin-gemcitabine)as a first-line therapy for locally advanced unresectable iCCA.The results showed promising outcomes,including a median OS of 22 mo and a 22%achievement in down-staging the tumor.In conclusion,TARE represents a viable treatment option for unresectable iCCA,and its combination with systemic chemotherapy has shown promising results.However,it is important to consider treatment-independent factors that can influence prognosis.Further research is necessary to identify optimal treatment combinations and predictive factors for a favorable response in iCCA patients.

Intrahepatic cholangiocarcinoma in patients with primary sclerosing cholangitis and ulcerative colitis: Two case reports

BACKGROUND Primary sclerosing cholangitis(PSC)is an extraintestinal manifestation of ulcerative colitis(UC).PSC is a well-known risk factor for intrahepatic cholangiocarcinoma(ICC),and ICC is known to have a poor prognosis.CASE SUMMARY We present two cases of ICC in patients with PSC associated with UC.In the first case,a tumor was found by magnetic resonance imaging(MRI)in the liver of a patient with PSC and UC who presented to our hospital with right-sided rib pain.The second patient was asymptomatic,but we unexpectedly detected two liver tumors in an MRI performed to evaluate bile duct stenosis associated with PSC.ICC was strongly suspected by computed tomography and MRI in both cases,and surgery was performed,but unfortunately,the first patient died of ICC recurrence 16 mo postoperatively,and the second patient died of liver failure 14 mo postoperatively.CONCLUSION Careful follow-up of patients with UC and PSC with imaging and blood tests is necessary for early detection of ICC.

Risk factors of intrahepatic cholangiocarcinoma in patients with hepatolithiasis:a case-control study

BACKGROUND:Why 3.3% to 10% of all patients with hepatolithiasis develop intrahepatic cholangiocarcinoma (ICC) remains unknown.We carried out a hospital-based case-control study to identify risk factors for the development of ICC in patients with hepatolithiasis in China.METHODS:Eighty-seven patients with pathologically diagnosed hepatolithiasis associated with ICC and 228 with hepatolithiasis alone matched by sex,age (±2 years),hospital admittance and place of residence were interviewed during the period of 2000-2008.Odds ratios (OR) and 95% confidence intervals (CI) were calculated for each risk factor.RESULTS:Among the patients with hepatolithiasis associated with ICC,the mean age was 57.7 years and 61.0% were female Univariate analysis showed that the significant risk factors for ICC development in hepatolithiasis were smoking,family history of cancer,appendectomy during childhood (under age 20),and duration of symptoms >10 years.In multivariate stepwise logistic regression analysis,smoking (OR=1.931,95% CI:1.000-3.731),family history of cancer (OR=5.175,95% CI:1.216-22.022),and duration of symptoms >10 years (OR=2.348 95% CI:1.394-3.952) were independent factors.CONCLUSION:Smoking,family history of cancer and duration of symptoms >10 years may be risk factors for ICC in patients with hepatolithiasis.

Clinicopathologic features, diagnosis and surgical treatment of intrahepatic cholangiocarcinoma in 104 patients

BACKGROUND: The outcome of surgical treatment of pa- tients with intrahepatic cholangiocarcinoma (ICC) is poor. This study was designed to analyze the relationship between clinicopathologic features and the survival time after opera- tion. METHODS: The operation was performed in 104 patients with mass-forming type ICC at our hospital between No- vember 1996 and May 2000. Seventy-nine patients (76.0%) were followed up successfully. Sixteen clinicopathological variables including age, sex, history of chronic liver di- sease , HBsAg, operation, adjuvant therapy, ascites, lymph node metastasis, invasion of adjacent organs, tumor size, necrosis of tumor, envelope, intrahepatic metastasis, Inter- national Union Against Cancer (UICC) TNM staging, his- tology, and cirrhosis were selected for univariate and multi- variate analyses to evaluate their influence on the prognosis. RESULTS: The accumulative 1-, 3-, 5-year survival rates of the 79 patients were 49.4%, 17.3%, 9.6% respectively. Univariate analysis revealed that sex (P=0.0221), HBsAg (P=0.0115), operation (P=0.0042), adjuvant therapy (P= 0.0389), ascites (P=0.0001), invasion (P=0.0220), intra- hepatic metastasis (P=0.0000) and TNM stage (P= 0.0001) were related to survival time. Multivariate analysis revealed that HBsAg, ascites and TNM stage were signifi- cantly related to prognosis. CONCLUSION: Early diagnosis and treatment and major hepatectomy are essential to improving the results of surgi- cal treatment of ICC patients.

Effect of contrast-enhanced ultrasound on differential diagnosis of intrahepatic cholangiocarcinoma and arterial phase enhanced hepatic inflammatory lesions

Objective: To investigate differential diagnosis between intrahepatic cholangiocarcinoma (ICC) and arterial phase enhanced hepatic inflammatory lesions in patients without liver cirrhosis using contrast-enhanced ultrasound (CEUS). Methods: ICC and hepatic inflammatory lesions cases with CEUS and pathological diagnosis between Sep 2013 and Oct 2016 were investigated retrospectively. Imaging features of conventional ultrasound and CEUS were analyzed. The parameters of time intensity curve (TIC), including the arrival time, peak intensity (PI) in the lesions, the starting time for washout, and the intensity difference at 3 min (Delta I-3) after contrast agent infection between the lesion and the liver parenchyma, were compared between ICC and hepatic inflammatory lesions. Results: Twenty-five ICC and fifteen inflammatory patients were included in this study. Seventeen ICC (68.0%) and two inflammatory cases (13.3%) showed bile duct dilatation on conventional ultrasound. Using CEUS, three ICC cases (12.0%) were misdiagnosed as inflammatory lesions and three inflammatory lesions (20.0%) as ICC; two ICC (8.0%) and one inflammatory case (6.7%) could not be made definite diagnosis. Washout started at 34.5 +/- 3.5 s and 61.5 +/- 12.9 s for ICC and inflammatory lesions respectively (P < 0.001). The intensity difference between lesion and liver parenchyma at 3 min after contrast agent injection was 10.8 +/- 3.1 dB in ICC and 4.2 +/- 2.3 dB in inflammatory group (P < 0.001). The sensitivity and specificity differentiating ICC and inflammatory lesions were 76% and 87% if the cut-off value of the intensity difference was 7.7 dB. Conclusions: Combined with TIC analysis, and particularly with the characteristic of the early-starting and obvious washout in ICC, CEUS can be useful in differential diagnosis between hepatic inflammatory lesions and ICC.

Can contrast enhanced ultrasound differentiate intrahepatic cholangiocarcinoma from hepatocellular carcinoma?

BACKGROUND Hepatocellular carcinoma(HCC)and intrahepatic cholangiocarcinoma(ICC)differ in treatment and prognosis,warranting an effective differential diagnosis between them.The LR-M category in the contrast-enhanced ultrasound(CEUS)liver imaging reporting and data system(LI-RADS)was set up for lesions that are malignant but not specific to HCC.However,a substantial number of HCC cases in this category elevated the diagnostic challenge.AIM To investigate the possibility and efficacy of differentiating ICC from HCC classified in the LR-M category according to the CEUS LI-RADS.METHODS Patients with complete CEUS records together with pathologically confirmed ICC and LR-M HCC(HCC classified in the CEUS LI-RADS LR-M category)between January 2015 and October 2018 were included in this retrospective study.Each ICC was assigned a category as per the CEUS LI-RADS.The enhancement pattern,washout timing,and washout degree between the ICC and LR-M HCC were compared using theχ2 test.Logistic regression analysis was used for prediction of ICC.Receiver operating characteristic(ROC)curve analysis was used to investigate the possibility of LR-M criteria and serum tumor markers in differentiating ICC from LR-M HCC.RESULTS A total of 228 nodules(99 ICCs and 129 LR-M HCCs)in 228 patients were included.The mean sizes of ICC and LR-M HCC were 6.3±2.8 cm and 5.5±3.5 cm,respectively(P=0.03).Peripheral rim-like arterial phase hyperenhancement(APHE)was detected in 50.5%(50/99)of ICCs vs 16.3%(21/129)of LR-M HCCs(P<0.001).Early washout was found in 93.4%(93/99)of ICCs vs 96.1%(124/129)of LR-M HCCs(P>0.05).Marked washout was observed in 23.2%(23/99)of ICCs and 7.8%(10/129)of LR-M HCCs(P=0.002),while this feature did not show up alone either in ICC or LR-M HCC.Homogeneous hyperenhancement was detected in 15.2%(15/99)of ICCs and 37.2%(48/129)of LR-M HCCs(P<0.001).The logistic regression showed that rim APHE,carbohydrate antigen 19-9(CA 19-9),and alpha fetoprotein(AFP)had significant correlations with ICC(r=1.251,3.074,and-2.767,respectively;P<0.01).Rim APHE presented the best enhancement pattern for diagnosing ICC,with an area under the ROC curve(AUC)of 0.70,sensitivity of 70.4%,and specificity of 68.8%.When rim hyperenhancement was coupled with elevated CA 19-9 and normal AFP,the AUC and sensitivity improved to 0.82 and 100%,respectively,with specificity decreasing to 63.9%.CONCLUSION Rim APHE is a key predictor for differentiating ICC from LR-M HCC.Rim APHE plus elevated CA 19-9 and normal AFP is a strong predictor of ICC rather than LR-M HCC.Early washout and marked washout have limited value for the differentiation between the two entities.

Expression of HBx protein in hepatitis B virus-infected intrahepatic cholangiocarcinoma

BACKGROUND: Hepatitis B virus (HBV) is an etiological factor of intrahepatic cholangiocarcinoma (ICC), but the pathogenic mechanisms remain unclear. This study aimed to investigate the expression and possible role of HBx, an HBV- encoded potentially oncogenic protein, in HBV-infected ICC. METHODS: Tissue samples were obtained from 54 specimens of HBV-infected ICC. Forty-four specimens were of peripheral type and 10 hilar type. Formalin-fixed, paraffin-embedded sections of the specimens were immunohistochemically stained for HBx and p53. RESULTS: HBx expression was found in 70.4% (38/54) of the specimens, and it was more frequently seen in the peripheral type than in the hilar type (79.5% vs 30.0%, P=0.002). All three well-differentiated ICCs expressed HBx, whereas 76.9% (30/39) moderately-differentiated and 41.7% (5/12) poorly-differentiated ICCs had HBx expression (P=0.033). Patients with HBx expression had a significantly higher prevalence of elevated serum alpha-fetoprotein (P=0.033). p53 protein expression was found in 18 of 54 cases (33.3%), and was not correlated with that of HBx. CONCLUSIONS: HBx may contribute to the pathogenesis of ICC, particularly the peripheral type. p53 abnormality may not play a significant role in HBx-mediated oncogenicity during ICC carcinogenesis.

Radiofrequency Ablation for Postoperative Recurrences of Intrahepatic Cholangiocarcinoma

Objective: Most recurrent intrahepatic cholangiocarcinoma (RICC) lost the opportunity of radical resection while most nonsurgical management failed to prolong patients’ survival. The efficacy and safety of radiofrequency ablation (RFA) as a local treatment for recurrent hepatocellular carcinoma have been confirmed by many clinical studies. The purpose of this study was to evaluate the efficacy, long-term survival and complications of RFA for RICC. Methods: A total of 12 patients with 19 RICCs after radical resection were included in this study. The tumors were 1.9–6.8 cm at the maximum diameter (median, 3.2±1.6 cm). All patients were treated with ultrasound guided RFA. There were two RFA approaches including percutaneous and open. Results: A total of 18 RFA treatment sessions were performed. Ablation was successful (evaluated by 1-month CT after the initial RFA procedure) in 18 (94.7%) of 19 tumors. By a median follow-up period of 29.9 months after RFA, 5 patients received repeated RFA because of intrahepatic lesion recurrence. The median local recurrence-free survival period and median event-free survival period after RFA were 21.0 months and 13.0 months, respectively. The median overall survival was 30 months, and the 1- and 3-year survival rates were 87.5% and 37.5%, respectively. The complication rate was 5.6% (1/18 sessions). The only one major complication was pleural effusion requiring thoracentesis. Conclusion: This study showed RFA may effectively and safely manage RICC with 3-year survival of 37.5%. It provides a treatment option for these RICC patients who lost chance for surgery.

Clinicopathologic significance of slug expression in human intrahepatic cholangiocarcinoma

AIM:To explore the expression and function of slug,a transcriptional repressor,in human intrahepatic cholangiocarcinoma(IHCC)and identify its role in IHCC progression.METHODS:Expression of slug was detected in 36 cases of IHCC and 12 cases of normal intrahepatic bile ducts and liver parenchyma by immunohistochemistry.The patients were divided into low slug expression group(< 20%of carcinoma cells stained)and high slug expression group(≥20%of carcinoma cells stained).Slug expression was correlated with clinicopathological parameters of IHCC patients.The patients were defined as short-term survivors if their survival time was<12 mo and as longterm survivors if their survival time was≥12 mo.RESULTS:Slug was not expressed in normal liver epi-thelium samples,lowly expressed in 15 tissue samples (10-,5+)and highly expressed in 21 tissue samples (16++;5+++)from IHCC patients.The survival rate of patients with a low slug expression was 33.3%(n =5)and 66.7%(n=10),respectively.The survival rate of patients with a high slug expression was 61.9% (n=13)and 38.1%(n=8),respectively(P=0.02).Lymph node metastasis was found in 4(26.7%)out of the 15 patients with a low slug expression and in 14(66.7%)out of the 21 patients with a high slug expression,respectively.The incidence rate of lymph node metastasis increased with the increasing slug expression level(P=0.003),and higher in patients with a high slug expression than in those with a low slug expression.Slug expression did not significantly correlate with the tumor size and stage or histologic grade,or with the gender and age of patients.CONCLUSION:Slug expression is a novel prognostic marker for IHCC with lymph node metastasis.

Etiological and clinicopathologic characteristics of intrahepatic cholangiocarcinoma in young patients

AIM:To investigate the prevalence,risk factors,and clinicopathologic characteristics of intrahepatic cholangiocarcinoma(ICC)in young patients.METHODS:A retrospective analysis was performed in ICC patients referred to the Eastern Hepatobiliary Surgery Hospital in Shanghai,China.Among 317 consecutively enrolled patients,40 patients were aged ≤40 years(12.61%).We compared the risk factors and clinicopathologic characteristics of these patients(groupⅠ:n=40)with those aged>40 years(group Ⅱ:n=277).RESULTS:Group I had distinct features compared with groupⅡ,including a low frequency of hepatolithiasis(P=0.000);a high positive rate of serum hepatitis B surface antigen(P=0.000)and hepatitis B virus(HBV)associated cirrhosis(P=0.038);a high frequency ofα-fetoprotein(>400μg/L)(P=0.011);a low frequency of carbohydrate antigen 19-9(>37 U/mL)(P=0.017);and a high frequency of liver histological inflammation(P=0.002).Although there was no significant difference between the two groups in regards to hepatic schistosomiasis,alcohol-associated cirrhosis and cirrhosis due to other causes(P>0.05),they only occurred in the elderly group.CONCLUSION:The risk factors are significantly different between young and elderly ICC patients.HBV and HBV-associated cirrhosis are the most important risk factors for young ICC patients.

Sarcomatoid intrahepatic cholangiocarcinoma mimicking liver abscess: A case report

Sarcomatoid intrahepatic cholangiocarcinoma(SICC)is an extremely rare and highly invasive malignant tumor of the liver.To our knowledge,the imaging findings of sarcomatous cholangiocarcinoma have been rarely reported;and radiological features of this tumor mimicking liver abscess have not yet been reported.CASE SUMMARY We present a case of SICC mimicking liver abscess.The patient,a 43-year-old male,complained of repeated upper right abdominal discomfort and intermittent distension over a period of one month.Radiology examination revealed a huge focal lesion in the right liver.The lesion was hypointense on computed tomography with honeycomb enhancement surrounded by enhanced peripheral areas.It showed a hypo-signal on non-contrast T1-weighted images and a hypersignal on non-contrast T2-weighted images.Radiologists diagnosed the lesion as an atypical liver abscess.The patient underwent a hepatectomy.After surgery,he survived another 2.5 mo before passing away.A search of PubMed and Google revealed 43 non-repeated cases of SICC reported in 20 published studies.The following is a short review in order to improve the diagnostic and therapeutic skills in cases of SICC.CONCLUSION This report presents the clinical and radiological features of SICC and imaging features which showed hypovascularity and progressive enhancement.SICC can present as a multilocular cyst on radiological images and it is necessary to distinguish this lesion from an atypical abscess.Simple surgical treatment is not the best treatment option for this disease.

Successful treatment of stage ⅢB intrahepatic cholangiocarcinoma using neoadjuvant therapy with the PD-1 inhibitor camrelizumab:A case report

BACKGROUND The prognosis of intrahepatic cholangiocarcinoma(ICC) with lymph node metastasis is poor.The feasibility of surgery is not certain,which is a contraindication according to the National Comprehensive Cancer Network guidelines.The role of immunotherapy as a neoadjuvant therapy for ICC is not clear.We herein describe a case of ICC with lymph node metastasis that was successfully treated with neoadjuvant therapy.CASE SUMMARY A 60-year-old man with a liver tumor was admitted to our hospital.Enhanced computed tomography and magnetic resonance imaging revealed a spaceoccupying lesion in the right lobe of the liver.Multiple subfoci were found around the tumor,and the right posterior branch of the portal vein was invaded.Liver biopsy indicated poorly differentiated cholangiocytes.According to the American Joint Committee on Cancer disease stage classification,ICC with hilar lymph node metastasis(stage ⅢB) and para-aortic lymph node metastasis was suspected.A report showed that two patients with stage ⅢB ICC achieved a complete response(CR) 13 mo and 16 mo after chemotherapy with a PD-1 monoclonal antibody.After multidisciplinary consultation,the patient was given neoadjuvant therapy,surgical resection and lymph node dissection,and postoperative adjuvant therapy.After three rounds of PD-1 immunotherapy(camrelizumab) and two rounds of gemcitabine combined with cisplatin regimen chemotherapy,the tumor size was reduced.Therefore,a partial response was achieved.Exploratory laparotomy found that the lymph nodes of Group 16 were negative,and the tumor could be surgically removed.Therefore,the patient underwent right hemihepatectomy plus lymph node dissection.The patient received six rounds of chemotherapy and five rounds of PD-1 treatment postoperatively.After 8 mo of follow-up,no recurrence was found,and a CR was achieved.CONCLUSION Neoadjuvant therapy combined with surgical resection is useful for advanced-stage ICC.This is the first report of successful treatment of stage ⅢB ICC using neoadjuvant therapy with a PD-1 inhibitor.

Notch1 Drives the Formation and Proliferation of Intrahepatic Cholangiocarcinoma

The molecular mechanisms underlying the development of intrahepatic cholangiocarcinoma(ICC)are not clear yet.In this study,we investigated the involvement of Notch1 in the development of ICC.The cDNA microarray analysis showed that Notch1 expression was higher in ICC tissues than in normal biliary epithelial cells.Stable transfection of Notchl receptor intracellular domain(NICD1)by hydrodynamic tail vein injection induced ICC formation in mice.Western blotting confirmed that Notchl signaling was activated in human ICC cell lines and mouse ICC tissues.Silencing Notchl with specific short interfering RNA(siRNA)inhibited the proliferation of ICC cells.Flow cytometry and Western blotting indicated that apoptosis was induced in Notchl-silenced ICC cells compared with controls.Additionally,Notchl silencing was associated with the inhibition of hairy and enhancer of split-1(Hes1)and activation of the phosphatase and tensin homolog(PTEN)/p53 pathway.Taken together,these data suggest that Notchl drives ICC formation and proliferation;downregulation of Notchl induces apoptosis in ICC cells;Notchl signaling may serve as a novel therapeutic target for the treatment of ICC.

EYA4 gene functions as a prognostic marker and inhibits the growth of intrahepatic cholangiocarcinoma

Background:The molecular prognostic markers and carcinogenesis of intrahepatic cholangiocarcinoma(ICC) have not been well documented.The purpose of this study was to investigate the prognostic value of the eyes absent homolog 4(EYA4) gene in ICC and its biological effects on ICC growth in vitro and in vivo.Methods:One hundred twelve patients with ICC who underwent hepatectomy were enrolled in the study.EYA4 mRNA and EYA4 protein levels in ICC and adjacent non-tumoral tissues were evaluated using real-time quantitative polymerase chain reaction and immunohistochemical staining,respectively.EYA4 protein levels in ICC cells were determined using western blot analysis.The associations between EYA4 expression and clinicopathologic features of ICC were analyzed.To identify independent prognostic factors,univariate and multivariate analyses were performed.The biological effects of EYA4 on ICC cells were evaluated by establishing stable EYA4-overexpressing transfectants in vitro,and EYA4's effects on tumor growth were evaluated by intra-tumoral injection of EVA4-expressing plasmids in a NOD/SCID murine model of xenograft tumors.Results:ICC tissues had significantly lower EYA4 mRNA and protein levels compared with adjacent non-tumoral tissues(both P < 0.001).Univariate and multivariate analyses showed that EYA4 protein level,tumor number,adjacent organ invasion,lymph node metastasis,and tumor differentiation were independent prognostic factors for diseasefree survival and overall survival(all P < 0.05).In vitro,EYA4 overexpression inhibited tumor cell growth,foci formation,and cell invasiveness.In vivo,intra-tumoral injection of EYA4-expressing plasmids significantly inhibited ICC growth in the murine xenograft model compared with the control group(P < 0.05).Conclusion:EYA4 gene functioned as a molecular prognostic marker in ICC,and its overexpression inhibited tumor growth in vitro and in vivo.

Do the existing staging systems for primary liver cancer apply to combined hepatocellular carcinoma-intrahepatic cholangiocarcinoma?

Background:The incidence of combined hepatocellular carcinoma-intrahepatic cholangiocarcinoma(cHCC-ICC)is relatively low,and the knowledge about the prognosis of cHCC-ICC remains obscure.In the study,we aimed to screen existing primary liver cancer staging systems and shed light on the prognosis and risk factors for cHCC-ICC.Methods:We retrospectively reviewed 206 cHCC-ICC patients who received curative surgical resection from April 1999 to March 2017.The correlation of survival measures with the histological types or with tumor staging systems was determined and predictive values of tumor staging systems with cHCC-ICC prognosis were compared.Results:The histological type was not associated with overall survival(OS)(P=0.338)or disease-free survival(DFS)(P=0.843)of patients after curative surgical resection.BCLC,TNM for HCC,and TNM for ICC stages correlated with both OS and DFS in cHCC-ICC(all P<0.05).The predictive values of TNM for HCC and TNM for ICC stages were similar in terms of predicting postoperative OS(P=0.798)and DFS(P=0.191)in cHCC-ICC.TNM for HCC was superior to BCLC for predicting postoperative OS(P=0.022)in cHCC-ICC.Conclusion:The TNM for HCC staging system should be prioritized for clinical applications in predicting cHCC-ICC prognosis.

Prognostic factors of recurrent intrahepatic cholangiocarcinoma after hepatectomy:A retrospective study

BACKGROUND Intrahepatic cholangiocarcinoma(ICC)is a highly malignant tumour.Hepatectomy is an effective treatment for early ICC,but postoperative recurrence greatly affects patient survival.Studies on recurrent ICC after hepatectomy are lacking.AIM To investigate the clinical characteristics of patients with recurrent ICC after hepatectomy,analyse prognostic factors and explore diagnosis and treatment strategies.METHODS A retrospective analysis was performed on all ICC patients undergoing hepatectomy from January 2013 to August 2021.Patients with postoperative recurrence were selected according to the inclusion and exclusion criteria.Cumulative overall survival was plotted by the Kaplan-Meier method,and differences were assessed by univariate survival analysis using the log-rank test.Multivariate analysis of cumulative survival was performed using the Cox proportional risk model.RESULTS During the 8-year study period,103 patients underwent ICC-related hepatectomy,and 54 exhibited postoperative recurrence.The median disease-free survival(DFS)was 6 mo,the median overall survival(OS)was 9 mo,and the cumulative OS rates at 1,2 and 3 years after the operation were 40.7%,14.8%and 7.4%,respectively.The median OS after recurrence was 4 mo,and the cumulative OS rates at 1,2 and 3 years after recurrence were 16.1%,6.7%and 3.4%,respectively.Multivariate analysis showed that alcohol consumption[hazard ratio(HR)=4.64,95%confidence interval(CI):1.53-14.04,P=0.007]and DFS<6 mo(HR=3.47,95%CI:1.59-7.60,P=0.002)were independent risk factors for the cumulative survival of patients with recurrence,while treatment after recurrence(HR=0.21,95%CI:0.08-0.55,P=0.001)was an independent protective factor.The median OS time of patients receiving multimodality therapy after recurrence of ICC was 7 mo,which was significantly higher than that of patients receiving only local therapy(3 mo),patients receiving systematic therapy(4 mo)and patients receiving the best supportive therapy(1 mo).Patients with recurrent ICC who received multimodality therapy had a significantly better long-term survival after recurrence than those who did not(P=0.026).CONCLUSION The prognosis of patients with recurrence after ICC-related hepatectomy is poor.Alcohol consumption and DFS<6 mo are independent risk factors in terms of the cumulative survival of patients with recurrence,while treatment after recurrence is an independent protective factor.Multimodality therapy can effectively improve the prognosis of patients.

Benefit of everolimus in treatment of an intrahepatic cholangiocarcinoma patient with a PIK3CA mutation

Intrahepatic cholangiocarcinoma(ICC) is a relatively rare form of liver cancer with a poor prognosis. The therapeutic options for patients with advanced ICC are limited and usually ineffective. There is currently no approved targeted therapy for ICC, although accumulating evidence supports inhibition of the PI3K/Akt/m TOR signaling pathway as a promising therapeutic strategy in the treatment of ICC. Here, we report a patient with stage IV ICC harboring a PIK3 CA mutation who responded well to the m TOR inhibitor everolimus. Computed tomography and magnetic resonance imaging demonstrated shrinkage of the tumor and maintenance of a partial response for 6.5 mo after everolimus treatment as the best response. To the best of our knowledge, this is the first clinical case report in the literature of clinical benefit from everolimus treatment in an ICC patient with PIK3 CA mutation.

High tumor mutation burden indicates a poor prognosis in patients with intrahepatic cholangiocarcinoma

BACKGROUND Intrahepatic cholangiocarcinoma(ICC)is malignancies of the biliary duct system and constitutes approximately 10%-20%of all primary liver cancers.Tumor mutation burden(TMB)is a useful biomarker across many cancer types for the identification of patients who will benefit from immunotherapy.Despite the role of TMB in calculating the effectiveness and prognosis of immune checkpoint inhibitors has been confirmed in multiple human cancer types,the prognostic value of TMB in ICC patients is rare investigated.AIM To investigate the prognostic value of TMB in patients with ICC.METHODS Data of 412 patients with ICC were included in the study.TMB was calculated as the total number of somatic non-silent protein-coding mutations divided by the coding region.The Kaplan-Meier method was used to analyze overall survival(OS),and relapse free survival(RFS).The cut-off value of TMB was determined by time-dependent receiver operating characteristic(ROC)curve.Cox regression was performed for multivariable analysis of OS.The nomogram and calibration curve were analyzed to construct and evaluate the prognostic model.RESULTS In the analysis of the time-dependent ROC curve,we defined 3.1 mut/Mb as the cut-off value of TMB.The Kaplan-Meier plot revealed that patients with high TMB had poor OS(HR=1.47,P=0.002)and RFS(HR=1.42,P=0.035).Cox regression analysis also demonstrated that TMB was an independent risk predictor for ICC(HR=1.43,P=0.0240).Furthermore,independent prognostic factors of ICC included CA19-9(HR=1.78,P=0.0005),chronic viral hepatitis(HR=1.72,P=0.0468),tumor resection(HR=2.58,P<0.0001)and disease progression(metastatic disease vs.solitary liver tumor;HR=2.55,P=0.0002).The nomogram and calibration curve also indicated the effectiveness of the constructed prognostic model.CONCLUSION TMB was an independent prognostic biomarker in patients with ICC.Moreover,patients with ICC with high TMB had poor OS and RFS as compared to those with low TMB.

Elevated expression of Gsα in intrahepatic cholangiocarcinoma associates with poor prognosis

BACKGROUND:The stimulatory G protein α subunit(Gsα)plays important roles in diverse cell processes including tu morigenesis. Activating mutations in Gsα gene(GNAS) have been reported to be associated with poor prognosis in various human carcinomas. Furthermore, Gsα signaling is crucial in promoting liver regeneration by interacting with growth factor signaling, indicating that Gsα might play a promoting role in cancer development. However, little is known about the correlation between Gsα levels and clinicopathological pa rameters in intrahepatic cholangiocarcinoma(ICC). METHODS:We performed immunoblotting to examine the expression levels of Gsα and Ki67 proteins in tumor tissues and the corresponding adjacent tissues. A total of 74 pair of specimens resected from 74 ICC patients were examined. The association between Gsα levels and clinicopathological find ings and prognosis of the patients was evaluated.RESULTS:Western blotting demonstrated that the expression of Gsα was significantly higher in ICC tissues compared with that in their corresponding adjacent tissues. Gsα protein was highly expressed in about half of ICC tissues(48.6%, 36/74)while only 28.4%(21/74) of tumor adjacent tissues showed Gsα high expression(P=0.011). High Gsα expression in ICC was significantly associated with the numbers of tumor nodules(P=0.037) and lymph node metastases(P=0.010)Moreover, the level of Gsα was significantly and positivelycorrelated with Ki67 expression(P<0.001). In addition, the recurrence-free survival rate and overall survival rate in the Gsα high group were significantly lower than those in the Gsα low group(P=0.004 and P=0.005, respectively).CONCLUSIONS:High Gsα expression is correlated with poor prognosis in ICC patients. Gsα might serve as a potential prognostic indicator of ICC.