Lymph Node Metastasis Of Intrahepatic Cholangiocarcinoma Research Articles

Mobilization and activation of tumor-infiltrating dendritic cells inhibits lymph node metastasis in intrahepatic cholangiocarcinoma

Lymph node metastasis (LNM) facilitates distant tumor colonization and leads to the high mortality in patients with intrahepatic cholangiocarcinoma (ICC). However, it remains elusive how ICC cells subvert immune surveillance within the primary tumor immune microenvironment (TIME) and subsequently metastasize to lymph nodes (LNs). In this study, scRNA-seq and bulk RNA-seq analyses identified decreased infiltration of dendritic cells (DCs) into primary tumor sites of ICC with LNM, which was further validated via dual-color immunofluorescence staining of 219 surgically resected ICC samples. Tumor-infiltrating DCs correlated with increased CD8+ T cell infiltration and better prognoses in ICC patients. Mechanistically, β-catenin-mediated CXCL12 suppression accounted for the impaired DC recruitment in ICC with LNM. Two mouse ICC cell lines MuCCA1 and mIC-23 cells were established from AKT/NICD or AKT/YAP-induced murine ICCs respectively and were utilized to construct the footpad tumor LNM model. We found that expansion and activation of conventional DCs (cDCs) by combined Flt3L and poly(I:C) (FL-pIC) therapy markedly suppressed the metastasis of mIC-23 cells to popliteal LNs. Moreover, β-catenin inhibition restored the defective DC infiltration into primary tumor sites and reduced the incidence of LNM in ICC. Collectively, our findings identify tumor cell intrinsic β-catenin activation as a key mechanism for subverting DC-mediated anti-tumor immunity in ICC with LNM. FL-pIC therapy or β-catenin inhibitor could merit exploration as a potential regimen for mitigating ICC cell metastasis to LNs and achieving effective tumor immune control.

Prediction and treatment of lymph node metastasis of intrahepatic cholangiocarcinoma

Intrahepatic cholangiocarcinoma (ICC) is the second largest primary liver cancer, and lymph node metastasis of ICC is commonly observed. Lymph node metastasis not only affects the prognosis of patients, but also determines the choice of treatment regimen. This article summarizes related articles on ICC in China and globally and elaborates on the serological markers and imaging indicators for predicting lymph node metastasis of ICC, the controversy over the selection of lymph node resection and postoperative adjuvant treatment for patients eligible for radical resection, and the research advances in neoadjuvant therapy for patients with advanced ICC who cannot undergo surgery, so as to provide theoretical support for the treatment of ICC patients and the prediction of lymph node metastasis.

Significance of Carbohydrate Antigen 19-9 as a biomarker in Hepatocellular Carcinoma and Cholangiocarcinoma

Background & Aims: Combined hepatocellular - cholangiocarcinomas (cHCC-CCs) are rare malignancies representing less than 1% of all primary liver cancers. Correct preoperative diagnosis is desirable because the frequency of lymph node metastasis in ICC and cHCC-CC, making lymph node dissection a necessity if curative resection to be attempted. This study aimed to investigate the significance of elevated CA19-9 in suspecting a diagnosis of Intrahepatic Cholangiocarcinoma (ICC) “non-invasively” in patients with typical radiological features of HCC. Methods This cross-sectional study was conducted on 54 patients with typical radiological criteria of HCC and elevated CA19-9 level. And were classified into two groups I included 22 patients (40.74%) who were diagnosed as HCC, group II included 30 patients (55.56%) were diagnosed as ICC, and there were 2 patients (3.7%) were diagnosed as cHCC-CC. Tumor markers (AFP and CA19-9), dynamic study (Triphasic CT or Dynamic MRI) were done for all patients. Target liver biopsy was done for histopathology and immuno-histochemistry using specific monoclonal antibodies against Glypican-3, Hep-par1, CK-7, CK-19 and CK-20 were done. Results: There was a statistically significant difference between HCC and ICC as regard CA19-9 and Alpha-fetoprotein (AFP). CA19-9 and AFP cut-offs were ˃ 58.9 U/mL and ˂ 25.8 ng/mL, respectively favoring the diagnosis of ICC, with very high sensitivity and specificity. CA19-9 level was 176.3 and 156.7 U/mL while AFP level was 460 and 170 ng/mL in cHCC-CC cases, respectively. Conclusion: CA19-9 could be a diagnostic marker of ICC in cases of typical radiological criteria of HCC with elevated CA19-9.

Mutated EPHA2 is a target for combating lymphatic metastasis in intrahepatic cholangiocarcinoma.

Exploring the genetic aberrations favoring metastasis is important for understanding and developing novel strategies to combat cancer metastasis. It remains lack of effective treatment for the dismal prognosis of intrahepatic cholangiocarcinoma (ICC). Here, we aimed to study genetic alternations during lymph node metastasis of ICC and investigate potential mechanisms and clinical strategy focused on mutations. We performed whole-exome sequencing and transcriptome sequencing on samples from 30 ICC patients, including lymph node metastases from five of the patients. We identified the alterations of genetic pattern related to lymph node metastases of ICC. EPHA2, a member of the tyrosine kinase family, was found to be frequently mutated in ICC. Correlation analysis indicated that EPHA2 mutations were closely associated with lymph node metastasis of ICC. In vitro and in vivo experiments revealed that EPHA2 mutations could lead to ligand independent phosphorylation of Ser897, and promote lymphatic metastasis of ICC, in which NOTCH1 signaling pathway played an important role. In both in vitro assays and patient-derived xenografts, an inhibitor of Ser897 phosphorylation effectively suppressed the metastasis of ICC with mutated EPHA2. Our findings demonstrated that EPHA2 mutants may be an attractive therapeutic target for lymphatic metastasis of ICC.

Expression and significance of amplified in breast cancer 1 in intrahepatic cholangiocarcinoma

Objective To investigate the amplified in breast cancer 1 (AIB1) and epithelial mesenchymal transition (EMT) related molecular markers in intrahepatic cholangiocarcinoma (ICC) and its relationship with clinical pathology. Methods The expression of AIB1 was detected by immunohistochemistry in the tissue adjacent to carcinoma and ICC. Recation between AIB1 clinical pathology of ICC was analyzed. Results The results of immunohistochemistry showed that the expression of AIB1 protein in ICC tissue was significantly higher than that in adjacent tissues, and the expression level of N-cadherin and Vimentin in ICC tissue was significantly higher than that in adjacent tissues. There was no significant difference in AIB1 protein expression with age, sex, tumor diameter, distant metastasis and pathological typing. The lymph node metastasis of ICC cases in positive expression of AIB1 occurred was 48.71% (19/39), and lymph node metastasis of ICC in negative expression of AIB1 occurred was 24.32% (9/37), there was statistically significant (P=0.035). In addition, the lymph node metastasis of ICC cases in positive expression of N-cadherin occurred was 47.50% (19/40), and lymph node metastasis of ICC in negative expression of N-cadherin occurred was 25.00% (9/36), two with statistical difference (P=0.027). There was no significant difference in Vimentin protein expression with age, sex, tumor diameter, distant metastasis and pathological typing. Western blotting assay results showed that AIBl expression in cancer tissue with positive lymph node metastasis was significantly higher than that in cancer tissue with negative lymph node metastasis, the difference was statistically significant (P=0.034). Conclusion The expression of AIB1 in ICC is associated with ICC lymph node metastasis and epithelial mesenchymal transition, suggesting that AIB1 may promote the invasion and metastasis of ICC by inducing epithelial mesenchymal transition of ICC. Key words: Intrahepatic cholangiocarcinoma; Amplified in breast cancer 1; Epithelial mesenchymal transition; Invasion and metastasis

Pathological Confirmation of Para‐Aortic Lymph Node Status as a Potential Criterion for the Selection of Intrahepatic Cholangiocarcinoma Patients for Radical Resection with Regional Lymph Node Dissection

Para-aortic lymph node (PAN) metastasis traditionally has been defined as distant metastasis. Many studies suggest that lymph node metastasis in intrahepatic cholangiocarcinoma (ICC) is one of the strongest prognostic factors for patient survival; however, the status of the PAN was not examined separately from regional lymph node metastasis in these reports. Here, we investigated whether regional lymph node metastasis without PAN metastasis in ICC can be classified as resectable disease and whether curative resection can have a prognostic impact. Between 1998 and 2010, a total of 47 ICC patients underwent hepatic resection and systematic lymphadenectomy with curative intent. We routinely dissected the PANs and had frozen-section pathological examinations performed intraoperatively. If PAN metastases were identified, curative resection was abandoned. We retrospectively investigated the prognostic factors for patient survival after curative resection for ICC without PAN metastases, with particular attention paid to the prognostic impact of lymphadenectomy. Univariate analysis identified concomitant portal vein resection, concomitant hepatic artery resection, intraoperative blood loss, intraoperative transfusion, and residual tumor as significant negative prognostic factors. However, lymph node status was not identified as a significant prognostic factor. The 14 patients with node-positive cancer had a survival rate of 20 % at 5 years. Based on multivariate analysis, intraoperative transfusion was an independent prognostic factor associated with a poor prognosis (risk ratio = 4.161; P = 0.0056). Regional lymph node metastasis in ICC should be classified as resectable disease, because the survival rate after surgical intervention was acceptable when PAN metastasis was pathologically negative.

High expression of Dickkopf‐related protein 1 is related to lymphatic metastasis and indicates poor prognosis in intrahepatic cholangiocarcinoma patients after surgery

Dickkopf-related protein 1 (DKK1) has been reported involved in metastasis and invasion in several tumors. This study sought to investigate the prognostic value of DKK1 in intrahepatic cholangiocarcinoma (ICC) and its role in promoting ICC metastasis. Tissue microarrays of 138 ICC patient samples were employed to detect DKK1, vascular endothelial growth factor C (VEGF-C), and matrix metalloproteinase 9 (MMP9) expression using immunohistochemistry. The prognostic significances were assessed by Kaplan-Meier survival estimates. DKK1 expression was measured in an ICC cell line (HCCC-9810) and ICC tissues by immunofluorescence assay, quantitative real-time polymerase chain reaction, and western blot. Serum levels of DKK1 from 37 ICC patients were tested by enzyme-linked immunosorbent assay. The role of DKK1 in proliferation, migration, invasion, and gene expression regulation was assessed by DKK1 depletion using small interfering RNA. Multivariate analyses revealed that DKK1 was an unfavorable predictor for overall survival and time to recurrence. The prognostic significance was retained in ICC patients with low recurrence risk (P < .05). DKK1 expression was elevated in an ICC cell line, tumor samples, and patient sera. High levels of DKK1 in ICC tissues correlated with elevated MMP9, VEGF-C, and metastasis of hepatic hilar lymph nodes. DKK1 depletion caused a decrease in cell migration and invasiveness, and down-regulation of MMP9 and VEGF-C expression. DKK1 is a novel prognostic biomarker for ICC, and it enhances tumor cell invasion and promotes lymph node metastasis of ICC through the induction of MMP9 and VEGF-C. DKK1 may be a potential therapeutic target for ICC.

Prediction of the Lymph Node Status in Patients with Intrahepatic Cholangiocarcinoma: Analysis of 320 Surgical Cases

Purpose: This study was conducted to identify factors involved in lymph node metastasis (LNM) and evaluate their role in predicting LNM in clinically lymph node negative (clinical stage I–III) intrahepatic cholangiocarcinoma (ICC). Materials and Methods: We selected 320 patients who were diagnosed with ICC with no apparent clinical LNM (T1–3N0M0). Age, gender, tumor boundary, histological differentiation, tumor size, and carbohydrate antigen 19-9 value were the studied factors. Univariate and multivariate logistic analysis were conducted. Receiver operating characteristics curve analysis was used to test the predicting value of each factor and a test which combined the associated factors was used to predict LNM. Results: LNM was observed in 76 cases (76/320, 23.8%). Univariate and multivariate analysis showed that histological differentiation as well as tumor boundary and tumor size significantly correlated with LNM. The sensitivity and negative predictive value for LNM for the three factors when combined was 96.1 and 95% respectively. This means that 5% of the patients who did not have the risk factors mentioned above developed LNM. Conclusion: This model used the combination of three factors (low-graded histological differentiation, distinct tumor boundary, small tumor size) and they proved to be useful in predicting LNM in ICC with clinically lymph node negative cases. In patients with these criteria, lymph node dissection or lymph node irradiation may be omitted and such cases may also be good candidates for stereotactic body radiotherapy (SBRT).

The Clinicopathologic Significance of the Expression of Vascular Endothelial Growth Factor-C in Intrahepatic Cholangiocarcinoma

Vascular endothelial growth factor-C (VEGF-C) is known to be an important lymphangiogenetic factor. Lymphatic spread is a key prognostic factor in intrahepatic cholangiocarcinoma (ICC). We studied the expression of VEGF-C in ICC tissues to clarify its clinicopathologic significance. The expression of VEGF-C in surgical specimens obtained from 36 patients with ICC who underwent hepatic resection was examined by immunohistochemistry and Western blotting. Strong staining was defined as the presence of VEGF-C immunoreactivity in at least 50% of the tumor cells. Immunoreactivity in approximately 10% to approximately 50% of the tumor cells was considered as weak staining, and less than 10% as no staining. Of the 36 patients with ICC, 15 patients (41.7%) showed a strong positive result for VEGF-C. Eleven cases (30.6%) were negative and 10 cases (27.8%) showed weak immunoreactivity. VEGF-C expression was significantly correlated with lymph node metastasis (P = 0.032) and positive surgical margin (P = 0.03). Patients who had strong positive staining for VEGF-C showed significantly less favorable survival rates compared with patients who had negative or weak staining (P < 0.01). Other significant prognostic factors by univariate analysis were vascular invasion, lymph node involvement, and positive surgical margin. Multivariate analysis demonstrated that strong VEGF-C expression (P = 0.028) and vascular invasion (P = 0.021) were independent factors indicating poor prognosis. Our data suggest that VEGF-C expression serves as an independent and important prognostic factor in ICC patients, and it may play an important role in the lymph node metastasis of ICC.