Introduction

Cholangiocarcinoma (CCA), a highly aggressive solid tumor with a five-year survival rate of only 5-15%, presents a significant challenge in terms of treatment efficacy. Consequently, there is a pressing demand for the identification of a safe and efficacious therapeutic approach for CCA. Magnolol, a compound derived from the Chinese herbal medicine Magnolia officinalis, has garnered attention for its potential anti-cancer properties. Nonetheless, the precise mechanism underlying the interaction between magnolol and CCA remains ambiguous.

Aims & Methods

Aims: The aim of this research is to investigate the inhibitory effect of magnolol on the proliferation of cholangiocarcinoma and elucidate the underlying mechanism.
Methods: At the cellular level, a series of experiments were conducted utilizing CCA cell lines, including assessments such as cell viability assay, EdU assay, colony formation assay, Transwell assay, and wound-healing assay, to investigate the impact of magnolol on CCA. Furthermore, a zebrafish model of cholangiocarcinoma was developed to examine the effects of magnolol on proliferation, angiogenesis, metastasis, and apoptosis. Additionally, a subcutaneous xenograft tumor model was established in nude mice to explore the effects of magnolol in vivo. Lastly, bioinformatics analysis, molecular docking, and western blot were performed to investigate the anticancer mechanism of magnolol.

Results

This study, utilized cellular-level experiments to demonstrate the inhibitory effects of magnolol on the proliferation, migration, and invasion of CCA in a time- and dose-dependent manner. Additionally, a zebrafish model was established to confirm the efficacy of magnolol in suppressing tumor growth, angiogenesis, and metastasis, as well as inducing apoptosis in CCA cells. Moreover, magnolol demonstrates its anti-tumor efficacy at a lower concentration compared to gemcitabine. Furthermore, our study utilizing a subcutaneous xenograft tumor model illustrates that magnolol effectively inhibits the advancement of CCA with minimal toxicity. Mechanistically, magnolol was found to downregulate the JAK2/STAT3 signaling pathway.

Conclusion

Overall, our research verified that magnolol had promising anti-tumor effects by targeting the JAK2/STAT3 pathway, which laid a solid foundation for CCA management.

Disclosure

The authors declare that there are no conflicts of interest.

Introduction

The sensitivity and accuracy of diagnosis for extrahepatic cholangiocarcinoma (EHCC) can be greatly improved by Spyglass DS choledochoscope, which can realize direct observation and biopsy . Studies have shown that endoscopic radiofrequency ablation (RFA) could prolong stent patency and survival in patients with unresectable extrahepatic cholangiocarcinoma (EHCC) . However, there are few reports about Spyglass DS choledochoscopy and endoscopic RFA therapy in the same stage of diagnosis and treatmen for unresectable EHCC.

Aims & Methods

Aim: To investigate the feasibility and safety of Spyglass DS combined with endoscopic RFA)for unresectable EHCC on the same stage.
Methods A total of 90 patients with extrahepatic bile duct obstruction treated in Hangzhou First People's Hospital from January 2013 to January 2022 were divided into two groups according to the diagnosis and treatment process: conventional group: ERCP + cytobrush/ Spyglass DS cholangioscopy first, and ERCP + RFA again when the pathological results were positive; simultaneous group: ERCP was performed via Spyglass DS cholangioscopy to examine the bile duct and biopsy the lesion under direct vision, and RFA was performed on the same stage for patients identified as malignant tumor. The success rate of procedure, recovery of liver function, the number of ERCPs and the incidence of postoperative adverse events were compared between the two groups.

Results

The sensitivity of Spyglass DS choledochoscope biopsy for EHCC was 87.5% (35/40) significantly higher than that of the cell brush group (38.9% (14/36), and the difference was statistically significant (P<0.05); All patients in both groups successfully completed endoscopic RFA with a 100% of procedure success rate; there was no statistically significant difference in the improvement of bilirubin between the two groups (P>0.05); the mean number of ERCPs in simultaneous group was significantly less than that in conventional group (1.00 vs. 2.59, P<0.001), which was statistically different; the incidence of postoperative adverse events between the two groups was 14.7% (5/34) vs. 14.3% (5/35), with no statistical significance (P=0.889).

Conclusion

Spyglass direct vision diagnosis + biopsy combined with RFA for unresectable extrahepatic cholangiocarcinoma can significantly improve the diagnostic sensitivity, reduce the number of ERCPs, and not increase the incidence of postoperative adverse events, which is a safe and effective diagnosis and treatment method with a high cost-benefit ratio.

Disclosure

NO

Introduction

Cholangiocarcinoma (CCA) constitutes a heterogeneous group of epithelial malignancies that arise in the biliary tree. CCA is a very aggressive cancer and carries a poor prognosis¹. Furthermore, in view of the constant increase in its incidence and mortality in recent decades throughout the world, it represents a growing global health problem^1,2. CCAs are divided into three subtypes according to their anatomical site of origin: intrahepatic CCA (iCCA), perihilar (pCCA), and distal (dCCA). Specifically, iCCA is characterize by high intertumoral clinical-pathological and molecular heterogeneity, leading to an innovative histological classification approved by WHO (ICD-O-3.2), into small bile duct iCCA and large bile duct iCCA³. The molecular pathogenesis of iCCA is very intricate and involves multiple molecular networks: among them, the Hedgehog (Hh)^4,6 signaling pathway. Indeed, the Hh pathway plays an important role in many aspects of iCCA, such as tumor survival, proliferation, migration and EMT reprogramming⁵. The impact of innovative therapies on the survival of patients with iCCA is an ongoing area of investigation. Evidence on the pathogenetic role of Hh in iCCA implies the possibility of targeting this signaling pathway for therapeutic purposes in iCCA subtypes.

Aims & Methods

The aim of this study is to demonstrate the antitumor efficacy of a natural therapeutic compound, from an in-house library, that inhibits the Hh pathway in experimental models of human iCCA, in vitro, in established iCCA cell lines (CCLP1) and primary cell lines derived from small or large bile duct iCCA. The iCCA samples from resected specimens have been processed by mechanical and enzymatic digestion to obtain patient-derived primary cell lines. The effect of Glabrescione B (GlaB), a selective inhibitor of GLI1 (Hh downstream transcriptional factor), has been evaluated. The dose-dependent efficacy of GlaB [0, 1, 5, 10, 20, 50 μM] or hyaluronic acid (HA) nanohydrogel-encapsulated Glab [0, 0.5, 1, 5, 10 μM], have been evaluated by Trypan Blue Exclusion Test and MTS assay (cell viability), at different time points (24h, 48h, 72h, 96h). Its ability as inhibitor of Hh signaling pathway has been evaluated by Western Blot analysis. All experiments have been conducted in N.3 experimental replicates.

Results

Our research shows a dose-dependent and time-dependent reduction of cell proliferation by Trypan Blue Exclusion Test in all cell lines (p<0.05). The decrease already occurs at low concentrations and at 24-hour incubation (40-50%) and becomes more evident at higher concentrations and at 96-hour incubation (80-90%) (p<0.05). Likewise, at the protein level, Gli1 knockdown, in a dose and time-dependent manner, is demonstrated (p<0.05). MTS assays confirmed the cytotoxicity effect of GlaB at 20 μM and 50 μM (p<0.05). Therefore, we decided to pursue the study with lower concentrations (from 1 μM to 10 μM) capable of knocking down the cell viability by 50%. Thereafter, successful, and stable encapsulation of GlaB [0.5-10 μM] in HA nanohydrogel has been accomplished. Data showed a dose-dependent and time-dependent inhibitory activity of HA-encapsuled GlaB [0.5-10 μM] in all cell lines (p<0.05).

Conclusion

Aberrant Hh pathway activation is widely known to be closely related with the development and malignant progression of various human cancers. Preclinical experimental data denote that dysregulation of this pathway is a determinant of cholangiocarcinoma progression and invasiveness. The achievements of this study could lay the groundwork for pre-clinical studies of HA-encapsuled Glab in iCCA.

References

1. Banales JM. et al., Nat Rev Gastroenterol Hepatol. 2020 Sep;17(9):557-588. Epub 2020 Jun 30. PMID: 32606456; PMCID: PMC7447603.
2. Bertuccio P. et al., J Hepatol. 2019 Jul;71(1):104-114. Epub 2019 Mar 23. PMID: 30910538.
3. European Association for the Study of the Liver. Electronic address: easloffice@easloffice.eu; European Association for the Study of the Liver. EASL-ILCA Clinical Practice Guidelines on Intrahepatic Cholangiocarcinoma. J Hepatol. 2023 Mar 22:S0168-8278(23)00185-X. Epub ahead of print. PMID: 37084797.
4. Riedlinger D. et al., J Hepatobiliary Pancreat Sci. 2014 Aug;21(8):607-15. Epub 2014 Apr 15. PMID: 24733827.
5. Cigliano A. et al., Expert Opin Ther Targets. 2017 May;21(5):471-483.1310842. Epub 2017 Mar 30. PMID: 28326864.
6. Razumilava N. et al., J Hepatol. 2014 Mar;60(3):599-605. Epub 2013 Nov 14. Erratum in: J Hepatol. 2014 Jul;61(1):185. PMID: 24239776; PMCID: PMC3944428.

Introduction

Among the diagnostic methods for EHCC, cell brush is the conventional method for EHCC diagnosis, but the sensitivity of cell brush and routine biopsy is low. The sensitivity and accuracy of EHCC diagnosis can be improved by single oral choledochoscopy. Radiofrequency ablation is a common and effective treatment for unresectable EHCC, providing local tumor control. Single oral choledochoscopy combined with radiofrequency ablation can improve the positive rate of pathological diagnosis.

Aims & Methods

To investigate the feasibility and safety of treating nonresectable extrahepatic cholangiocarcinoma (EHCC) with single resection combined with radiofrequency ablation (RFA).
A total of 149 patients with suspected extrahepatic cholangiocarcinoma treated in Hangzhou First People's Hospital from January 2013 to January 2023 were reviewed. According to the treatment process, 114 patients were eventually included and divided into two groups (n=48 cases) : Single oral choledochoscopy was performed by ERCP and the lesion tissue was biopsied under direct vision. Intraoperative rapid pathologic examination was performed on the same stage RFA for the patients who were pathologically identified as malignant tumors. Regular times group (n = 66) : first by endoscopic retrograde pancreatic angiography (endoscopic retrograde cholangiopancreatography, ERCP) + cells brush or a single biopsy samples by mouth choledochoscope examination, pathological results to obtain positive line of ERCP + RFA again; The operation success rate, postoperative liver function recovery, average ERCP number, postoperative adverse event incidence, length of hospitalization and cost were compared between the two groups.

Results

1. Patients in both groups successfully completed the RFA, the operation success rate was 100% (114/114).
2. There was no significant difference in the overall incidence of postoperative adverse events between the two groups [39.58% (19/48) vs. 39.40% (26/66),χ2=0.00, P=0.984].
3. There was no significant difference in the proportion of patients with total bilirubin improvement between the two groups [41.67% (20/48) vs. 42.42% (28/66), χ2= 0.32, P=0.750].
4. The average number of ERCP in the same group was significantly lower than that in the conventional group, and the difference was statistically significant (1.00±0.00 times/person vs. 2.64±0.49 times/person, t=0.77, P<0.001).
5. The average length of hospital stay in the same group was significantly longer than that in the conventional group, with statistical significance (8.40±3.63 days vs. 17.47±9.82 days, t=6.21, P<0.001).
6. The average hospitalization cost in the same group was significantly higher than that in the conventional group, and the difference was statistically significant (27718.31±9142.02 yuan versus 49112.76±16153.14 yuan, t=7.38, P<0.001).

Conclusion

Single oral choledochoscopy combined with biopsy and RFA in the diagnosis and treatment of unresectable EHCC can reduce the number of operations and does not increase the incidence of postoperative adverse events, which is a safe and effective method with a high cost-benefit ratio.

References

[1] Le V H, O'Connor V V, Li D, et al. Outcomes of Neoadjuvant Therapy for Cholangiocarcinoma: A Review of Existing Evidence Assessing Treatment Response and R0 Resection Rate [J]. J Surg Oncol, 2021, 123(1): 164-71.
[2] Chen R, Zheng D, Li Q, et al. Immunotherapy of Cholangiocarcinoma: Therapeutic Strategies and Predictive Biomarkers [J]. Cancer Lett, 2022, 546: 215853.
[3] Rizzo A, Ricci A D, Brandi G. Durvalumab: An Investigational Anti-Pd-L1 Antibody for the Treatment of Biliary Tract Cancer [J]. Expert Opin Investig Drugs, 2021, 30(4): 343-50.
[4] Sadeghi S. Infigratinib for Cholangiocarcinoma [J]. Drugs Today (Barc), 2022, 58(7): 327-34.
[5] Vignone A, Biancaniello F, Casadio M, et al. Emerging Therapies for Advanced Cholangiocarcinoma: An Updated Literature Review [J]. J Clin Med, 2021, 10(21).
[6] Jang D K, Kim J, Paik C N, et al. Endoscopic Retrograde Cholangiopancreatography-Related Adverse Events in Korea: A Nationwide Assessment [J]. United European Gastroenterol J, 2022, 10(1): 73-9.
[7] Hollenbach M, Hoffmeister A. Adverse Events in Endoscopic Retrograde Cholangiopancreaticography (Ercp): Focus on Post-Ercp-Pancreatitis [J]. United European Gastroenterol J, 2022, 10(1): 10-1.

Disclosure

The authors declare that there are no conflicts of interest.