Comparison of Staging Systems of Hepatocellular Carcinoma

Many staging systems of hepatocellular carcinoma (HCC) were established; however, there is no consensus on which is proper in predicting prognosis. This study aims to evaluate various commonly used staging systems of HCC. Patients who underwent surgery during 2001–2007 were included. All patient data were retrospectively staged using six staging systems, that are American Joint Committee on Cancer (AJCC) Tumour-Node-Metastasis (TNM), Okuda staging, Cancer of the Liver Italian Program (CLIP), Barcelona Clinic Liver Cancer (BCLC), Chinese University Prognostic Index (CUPI), and Japan Integrated Staging (JIS). Child-Pugh classification was also evaluated. The staging systems were compared by mean of overall and disease-free survival. Total of 99 patient data were enrolled in the analyses. All staging systems except Okuda were significant in determining overall survival in univariate analyses. In multivariate analyses, TNM and Child-Pugh demonstrated better predictive power for overall survival. In terms of disease-free survival, univariate analyses revealed that TNM, CLIP, BCLC, CUPI, and JIS were significant, and TNM was the best predictive staging system in multivariate analyses. In our study, TNM and Child-Pugh are the representative systems in predicting survival of HCC patients who undergo surgical resection. Moreover, they are practical and easily assessable in clinical practice

UPLC-ESI-MRM/MS for Absolute Quantification and MS/MS Structural Elucidation of Six Specialized Pyranonaphthoquinone Metabolites From Ventilago harmandiana

Pyranonaphthoquinones (PNQs) are important structural scaffolds found in numerous natural products. Research interest in these specialized metabolites lies in their natural occurrence and therapeutic activities. Nonetheless, research progress has thus far been hindered by the lack of analytical standards and analytical methods for both qualitative and quantitative analysis. We report here that various parts of Ventilago harmandiana are rich sources of PNQs. We developed an ultraperformance liquid chromatography-electrospray ionization multiple reaction monitoring/mass spectrometry method to quantitatively determine six PNQs from leaves, root, bark, wood, and heartwood. The addition of standards in combination with a stable isotope of salicylic acid-D-6 was used to overcome the matrix effect with average recovery of 82% +/- 1% (n = 15). The highest concentration of the total PNQs was found in the root (11,902 mu g/g dry weight), whereas the lowest concentration was found in the leaves (28 mu g/g dry weight). Except for the root, PNQ-332 was found to be the major compound in all parts of V. harmandiana, accounting for similar to 48% of the total PNQs quantified in this study. However, PNQ-318A was the most abundant PNQ in the root sample, accounting for 27% of the total PNQs. Finally, we provide novel MS/MS spectra of the PNQs at different collision induction energies: 10, 20, and 40 eV (POS and NEG). For structural elucidation purposes, we propose complete MS/MS fragmentation pathways of PNQs using MS/MS spectra at collision energies of 20 and 40 eV. The MS/MS spectra along with our discussion on structural elucidation of these PNQs should be very useful to the natural products community to further exploring PNQs in V. harmandiana and various other sources

The microRNA-15a-PAI-2 axis in cholangiocarcinoma-associated fibroblasts promotes migration of cancer cells

Background: Cholangiocarcinoma (CCA) has an abundance of tumor stroma which plays an important role in cancer progression via tumor-promoting signals. This study aims to explore the microRNA (miRNA) profile of CCA-associated fibroblasts (CCFs) and the roles of any identified miRNAs in CCA progression. Methods: miRNA expression profiles of CCFs and normal skin fibroblasts were compared by microarray. Identified downregulated miRNAs and their target genes were confirmed by real-time PCR. Their binding was confirmed by a luciferase reporter assay. The effects of conditioned-media (CM) of miRNA mimic- and antagonist-transfected CCFs were tested in CCA migration in wound healing assays. Finally, the levels of miRNA and their target genes were examined by real-time PCR and immunohistochemistry in clinical CCA samples. Results: miR-15a was identified as a downregulated miRNA in CCFs. Moreover, PAI-2 was identified as a novel target gene of miR-15a. Recombinant PAI-2 promoted migration of CCA cells. Moreover, CM from miR-15a mimic-transfected CCFs suppressed migration of CCA cells. Lower expression of miR-15a and higher expression of PAI-2 were observed in human CCA samples compared with normal liver tissues. Importantly, PAI-2 expression correlated with poor prognosis in CCA patients. Conclusions: These findings highlight the miR-15a/PAI-2 axis as a potential therapeutic target in CCA patients

Analysis of clonality and vascular biology in Hepatocellular carcinoma

Hepatocellular carcinoma (HCC) arising in cirrhosis is frequently multifocal. Whether HCC develops monoclonally or multiclonally is an unresolved question. Of the multiple tumour nodules which often develop in cirrhotic, it has not been established whether the smaller lesions represent intrahepatic metastases or de novo cancers. The degree of genomic heterogeneity was therefore assessed, and the clonal evolution was studied in synchronous HCCs in cirrhosis from patients undergoing elective orthotopic liver transplantation. All samples studied were negative on p53 immunohistochemistry, and no mutation in exons 5-9 of the p53 gene was found by performing polymerase chain reaction and direct DNA sequencing analysis. Using arbitrarily primed-polymerase chain reaction (AP-PCR) technique, reproducible and interpretable fingerprinting patterns of amplified genomic DNA isolated from microdissected paraffin-embedded tissues were generated. The fingerprints were highly polymorphic amongst HCCs and regenerative nodules. Clonal evolution was found to occur in HCCs more than 6 mm in diameter which consistently contain multiple clones. In addition, the genomic fingerprint patterns of the extra-hepatic metastatic lesions were polymorphic compared to those of the corresponding primary HCCs. By contrast, DNA fingerprints of multiple areas of primary fibrolamellar carcinomas (FLCs) and all their metastatic lesions were identical. The advent of laser capture microdissection (LCM) technology allowed analysis of selected cell groups within a single tumour much more rapid and simple to apply. The control of neovascularisation in tumours, particularly in HCCs, depends on a net balance of positive and negative angiogenic factors. Transfection of thrombospondin-1, an important negative factor, into Sk-Hep-1 cells was associated with decreased thymidine incorporation in vitro and reduced primary tumour growth in vivo. Antiangiogenic therapy after removal of a dominant HCC might be a useful treatment to suppress the growth of residual or synchronous tumours

Surgical Treatment of Renal Cell Carcinoma with Inferior Vena Cava Thrombus: Using Liver Mobilization Technique to Avoid Cardiopulmonary Bypass

To evaluate the results of surgical treatment of renal cell carcinoma (RCC) with inferior vena cava (IVC) thrombus and describe the use of a transabdominal approach with liver mobilization to avoid cardiopulmonary bypass (CPB). METHODS: From February 2002 to January 2006, 109 patients with RCC were surgically treated at Siriraj Hospital. Twelve patients had an IVC thrombus, infrahepatic (level I), retrohepatic (level II), suprahepatic (level III) and intra-atrial (level IV) in one, two, eight and one patient, respectively. Patients' characteristics, pathological features, survival and morbidity were evaluated. RESULTS: Mean age was 58 years (range, 37–74 years). CPB was used in one patient with level IV thrombus. All patients (92%) with level I–III IVC thrombi underwent successful removal by transabdominal approach without any form of bypass. Mean operative time was 302 minutes (range, 195–420 minutes). The mortality rate was 16% (2 of 12) with sepsis and pulmonary embolism. One patient had colonic injury requiring primary repair. At the mean follow-up of 17 months (range, 3–35 months), of 10 patients, one died due to distant metastases, two were lost to follow-up and seven (60%) were still alive. Five patients (42%) were disease-free at the last follow-up. CONCLUSION: These results support the aggressive surgical removal of RCC with IVC thrombus as the initial treatment. Most of the thrombi can be approached and safely controlled by a transabdominal approach without any form of bypass. Tumour thrombus removal provides a high survival chance and offers improvement in quality of life

Study of siRNA Delivery via Polymeric Nanoparticles in Combination with Angiogenesis Inhibitor for The Treatment of AFP-Related Liver Cancer

Angiogenesis inhibitor drugs have been explored as important pharmacological agents for cancer therapy, including hepatocellular carcinoma. These agents have several drawbacks, such as drug resistance, nonspecific toxicity, and systemic side effects. Therefore, combination therapy of the drug and small interfering RNA could be a promising option to achieve high therapeutic efficacy while allowing a lower systemic dose. Therefore, we studied adding an alpha-fetoprotein siRNA (AFP-siRNA) incorporated on polymeric nanoparticles (NPs) along with angiogenesis inhibitor drugs. The AFP siRNA-loaded NPs were successfully synthesized at an average size of 242.00 ± 2.54 nm. Combination treatment of AFP-siRNA NPs and a low dose of sunitinib produced a synergistic effect in decreasing cell viability in an in vitro hepatocellular carcinoma (HCC) model. AFP-siRNA NPs together with sorafenib or sunitinib greatly inhibited cell proliferation, showing only 39.29 ± 2.72 and 44.04 ± 3.05% cell viability, respectively. Moreover, quantitative reverse transcription PCR (qRT-PCR) demonstrated that AFP-siRNA incorporated with NPs could significantly silence AFP-mRNA expression compared to unloaded NPs. Interestingly, the expression level of AFP-mRNA was further decreased to 28.53 ± 5.10% when sunitinib was added. Therefore, this finding was considered a new promising candidate for HCC treatment in reducing cell proliferation and enhancing therapeutic outcomes