High AFP Level Research Articles

Expression of Pregnancy Up-regulated Non-ubiquitous Calmodulin Kinase (PNCK) in Hepatocellular Carcinoma.

Pregnancy up-regulated non-ubiquitous calmodulin kinase (PNCK) is a member of calmodulin kinase, and overexpression of PNCK with involvement in carcinogenesis have been reported in HER-2 amplified breast cancer, clear cell renal cell carcinoma and nasopharygeal carcinoma. However, the expression of PNCK and its clinical implication have not been elucidated in hepatocellular carcinoma (HCC). We investigated PNCK expression at both the protein and mRNA level using immunohistochemistry (IHC) and microarray gene expression profiling in HCC tissue samples, and evaluated its association with clinicopathological parameters and their potential prognostic significance. High PNCK protein expression and high PNCK mRNA level was observed in 61.7% and 34.7% of total HCC cases, respectively. PNCK mRNA level was higher in tumor tissues than in background non-tumor tissues, and significantly correlated with protein expression by IHC. High PNCK expression was associated with higher Edmondson grade, intrahepatic metastasis, microvascular invasion and higher AFP levels. Patients with high PNCK expression showed shorter recurrence-free survival and disease-specific survival, and high mRNA expression of PNCK was an independent prognostic factor in disease-specific survival. Up-regulation of PNCK expression as well as its association with poor prognosis was demonstrated in HCC. PNCK might be a prognostic biomarker of HCC, and could be a potential candidate therapeutic target.

The expression level and diagnostic value of microRNA-22 in HCC patients

Background Involvements of microRNA-22 (miR-22) in cancer have attracted much attention, but its role in diagnosis of hepatocellular carcinoma (HCC) is still largely unknown. Therefore, the aim of this study was to investigate the expression level and the prognostic value of miR-22 in HCC patients. Methods Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to evaluate serum level of miR-22 in 108 HCC patients and 67 healthy controls. The relationship between miR-22 expression level and clinicopathologic characteristics was analysed via chi-square test. Receiver operating characteristic (ROC) curve was built to estimate the diagnostic value of serum miR-22 in HCC. Results miR-22 expression was significantly down-regulated in HCC compared to that in healthy controls (p < .05). And the low miR-22 expression was significantly associated with vein invasion (p = .002), TNM stage (p = .013) and high serum levels of AFP (α-fetoprotein), ALT (alanine aminotransferase), AST (aspartate aminotransferase) and ALP (alkaline phosphatase. miR-22 had a high diagnostic value with area under the curve of 0.866 corresponding with a sensitivity of 89.3% and a specificity of 68.9%, respectively. Conclusion miR-22 expression was down-regulated in HCC patients. Serum miR-22 might be a novel diagnostic marker in HCC.

Genomic Profiles of a Patient of Pulmonary Hepatoid Adenocarcinoma With High AFP Level: A Case Report

Hepatoid adenocarcinoma of lung (HAL) is a rare and aggressive tumor. The current study reported a new HAL case in the right lower lung with high serum α-fetoprotein (AFP) level in a 71-year-old male patient. After the confirmation of morphology and immunohistochemistry, the patient was diagnosed clinically with HAL and treated with radio-frequency ablation. However, the patient whose disease progressed eventually died 4 months after diagnosis. Whole genome sequencing analysis identified a driver gene mutation in the FAT atypical cadherin 1 gene (FAT1) and the copy number loss. The tumor was microsatellite-stable and tumor mutation burden (TMB) was 1.69 mutations/Mb. PD-L1 expression was negative by IHC. Our finding provide further clues for the molecular basis of HAL and the efficacy of immunotherapy needs to be explored.

Preoperative elevated plasma fibrinogen level predicts tumor recurrence and poor prognosis in patients with hepatocellular carcinoma.

Elevated plasma fibrinogen has been reported to be associated with poor prognosis in several cancers. The aim of present study was to investigate the prognostic value of preoperative plasma fibrinogen in hepatocellular carcinoma (HCC) patients. Data were collected retrospectively from 302 HCC patients who received hepatectomy. The association between fibrinogen and clinicopathological characteristics was evaluated. Both univariate and multivariate analyses were performed to identify prognostic factors for disease-free survival (DFS) and overall survival (OS). And accordingly, the nomograms were constructed. Elevated plasma fibrinogen (>4 g/L) was correlated with larger tumor diameter, the presence of vascular invasion, lower MELD score, higher NLR, advanced Barcelona Clinic Liver Cancer stage and poor-moderate pathological differentiation. On multivariate analysis, the elevated plasma fibrinogen was found independently associated with poor DFS (HR =1.575, P=0.024) and OS (HR =2.051, P=0.025). And the nomograms including fibrinogen were constructed to predict DFS and OS for HCC patients. Both DFS and OS in patients with plasma fibrinogen >4 g/L were significantly lower than those with fibrinogen ≤4 g/L (1-, 3-, 5-year DFS: 34.2%, 19.5% and 0.0% vs. 60.4%, 34.2% and 30.2%; 1-, 3-, 5-year OS: 83.4%, 62.7% and 48.8% vs. 95.4%, 84.3% and 75.8%, both P<0.001). Besides, subgroup analyses also showed the prognostic values of fibrinogen in HCC patients with/without cirrhosis or high AFP levels, and in those with single tumor and BCLC 0-A stage. Preoperative elevated plasma fibrinogen was an independent prognostic factor associated with poor prognosis in HCC patients receiving liver resection.

1762 A Rare Case of an Alpha-Fetoprotein-Producing Esophageal Adenocarcinoma

INTRODUCTION: AFP-producing esophageal adenocarcinoma (EAC) is a very rare occurrence. Elevation of serum AFP is commonly associated with hepatocellular carcinoma and yolk sac tumors, but rarely with esophageal carcinoma. This is a case of a 75-yr-old female who presented with progressive weakness and decrease in appetite, she was diagnosed with EAC despite high suspicion for HCC due to the extremely elevated AFP level. Prognosis is poor given the advanced presenting stage and high metastatic potential when finally diagnosed. CASE DESCRIPTION/METHODS: A 75-yr-old female with a past medical history of gastroesophageal reflux disease and Barrett’s esophagus status-post multiple upper endoscopies and a colonoscopy presented with progressive weakness, decrease in appetite, and abdominal pain over a few weeks. She also complained of dysphagia for solid food, and recent weight loss. On physical exam, she had mild epigastric tenderness. Her labs revealed markedly elevated CEA of 742, alpha-fetoprotein 46,135, and CA 19-9 of 6842. In the hospital, she had tachycardia so a CTA of her lungs was done to rule out a pulmonary embolism but instead revealed gastric thickening, celiac adenopathy, and multiple large liver masses. These findings were confirmed by a CT of her abdomen and pelvis with and without IV and oral contrast. Upper endoscopy revealed a large non-obstructing mass in the distal esophagus that extended into the stomach. A needle aspiration biopsy of a liver lesion revealed esophageal adenocarcinoma with metastasis. DISCUSSION: There have been only fifteen reported cases of AFP-producing esophageal adenocarcinoma in the literature. Our patient presented not only with EAC but also extremely high levels of AFP &gt;40,000, with 320 ng/mL as the highest reported AFP level previously. Most EACs were located over the lower third of the esophagus and were related to Barrett’s esophagus as seen in our patient. The serum AFP level can be serially measured for monitoring clinical status, evaluating cure, and assessing recurrence or metastases. Most cases were treated with adjuvant chemotherapy with cisplatin plus 5-fluorouracil (5-FU) as first-line and combination of paclitaxel and cisplatin as second-line. Our patient was treated with oxaplatin, folinic acid, and 5-FU. She had temporary reduction of metastasis, but unfortunately there was recurrence. Clinicians should consider AFP-producing EAC in their differential diagnosis when working up a liver mass in the setting of elevated AFP or liver function impairment.

DEPTOR induces a partial epithelial-to-mesenchymal transition and metastasis via autocrine TGF\u03b21 signaling and is associated with poor prognosis in hepatocellular carcinoma

BackgroundDEPTOR is an endogenous inhibitor of mTORC1 and mTORC2 that plays a vital role in the progression of human malignances. However, the biological function of DEPTOR in HCC metastasis and the underlying molecular mechanisms are still unclear.MethodsWestern blot analysis and immunohistochemistry(IHC) were employed to examine DEPTOR expression in HCC cell lines and tissues. A series of in vivo and in vitro assays were performed to determine the function of DEPTOR and the possible mechanisms underlying its role in HCC metastasis.ResultsWe found that DEPTOR was frequently overexpressed in HCC tissues, and its high expression was associated with high serum AFP levels, increased tumor size, vascular invasion and more advanced TMN and BCLC stage, as well as an overall poor prognosis. Functional experiments demonstrated that DEPTOR silencing inhibited the proliferation and mobility of HCC cells in vitro and suppressed tumor growth and metastasis of HCC cells in vivo. Accordingly, DEPTOR overexpression promoted the invasion and metastasis of HCC cells in vitro and in vivo, but had no effect on cell proliferation in vitro. Overexpression of DEPTOR induced EMT by snail induction. Conversely, knockdown of snail expression impaired the DEPTOR-induced migration, invasion and EMT of HCC cells. Furthermore, we found that the increase of snail expression by DEPTOR overexpression was due to an activation of TGF-β1-smad3/smad4 signaling possibly through feedback inhibition of mTOR.ConclusionDEPTOR promotes the EMT and metastasis of HCC cells by activating the TGF-β1-smad3/smad4-snail pathway via mTOR inhibition. Therefore, targeting DEPTOR may be an ideal treatment strategy for inhibiting the growth and metastasis of HCC.

Increase in Incidence of Hepatocellular Carcinoma in Chronic Hepatitis C Chinese After Sustained Virologic Response with Direct Acting Antiviral Agents as Compared to Pegylated Interferon Plus Ribavirin Therapy: A Prospective Open-Label Study

Background: The incidence of hepatocellular carcinoma (HCC) decreased significantly in persons with chronic hepatitis C (CHC) who achieved sustained virologic response (SVR) after pegylated-interferon plus ribavirin (PR) therapy. It remains controversial whether there is any difference in HCC incidence after SVR using direct-acting antiviral agents (DAAs), as compared to PR therapy. We report a prospective observational study which compare the SVR and post-SVR HCC incidence in CHC Chinese who had completed either DAAs or PR therapy. Methods: This prospective observational study (NCT02578693) was conducted at two sites in Beijing and Hong Kong, to evaluate SVR in CHC Chinese with completion of DAAs or PR therapy. Those with SVR were followed up at 12-24 weekly intervals with surveillance for HCC performed by ultrasonography and alpha-fetoprotein at each visit. Multivariate Cox analysis was used to explore factors associated with post-SVR occurrence of HCC. Findings: Between October 2015 and May 2017, SVR was observed in 519 of the 533 (97.4%) and 817 of 1202 (68.0%) CHC patients with completion of anti-HCV therapy (p = 0.0001), respectively. Subjects that achieved SVR were followed with a median duration of 3 years. During the follow-up period, HCC developed in 28 (5.8%) and 25 (3.3%) post-SVR patients treated with DAAs and PR therapy, respectively (p = 0.036). The use of DAAs, older age (≥55 years), diabetes mellitus, higher LSM (≥17.5 kPa), and higher AFP level (≥30 ng/ml) at SVR were associated with increased risk of HCC (hazard ratios: 1.9, 3.1, 3.9, 4.9 and 5.2 respectively). Interpretation: After a median follow-up of 3 years, a lower incidence of HCC was observed in CHC patients who had SVR with PR therapy compared to those treated with DAAs. HCC surveillance in CHC patients with SVR should be performed more stringently in those with high risk factors and treated with DAAs. Clinical Trial Registration: The clinical protocol was registered at ClinicalTrials.gov (NCT02578693). Funding Statement: This is an investigator-initiated study supported by Chinese medical association (Grant No.13071110496) and capital characteristic clinic project of the Beijing municipal science and technology commission (Grant No. Z181100001718034). RFS is supported by the Emory Center for AIDS Research NIH grant 2P30-AI-050409. Declaration of Interests: The authors declare no conflict of interest. Ethics Approval Statement: Approval was obtained from the Ethics Committees in Hong Kong (Hong Kong Clinical Research Ethics Committee, HKCREC) and Beijing (Ethics Committee of Beijing 302 hospital). Informed consent was obtained from all patients and the study was conducted in compliance with the 1975 Declaration of Helsinki, Good Clinical Practice guidelines, and local regulatory requirements.

Clinical and prognostic significance of serum levels of fatty acid binding proteins in hepatocellular carcinoma (HCC).

4099 Background: Limited data are available about the prognostic effect of fatty acid binding proteins (FABP) in viral and non-viral-related hepatocellular carcinoma (HCC). Previous studies suggested that selected FABP could be a potential target markers for HCC chemotherapy response and may correlated with presence of cirrhosis and poor outcome. We aimed to test the association between plasma levels of Liver (L)-FABP, Heart (H)-FABP, and Adipose (A) FABP and HCC. Methods: we enrolled 767 HCC patients from MD Anderson Cancer Center. Under IRB approval, baseline patients’ characteristics were retrieved from medical records and blood samples were collected and tested form plasma levels of L-, A-, H-, FABPs. Descriptive statistics were performed and the median values of FABPs among 200 normal controls (NC) were used as cutoff values of FABPs. Overall survival (OS) was estimated by Kaplan Meier curve and log rank test. Results: FABPs were highly expressed in HCC cases than controls. Mean values (±SE) of AFABP, HFABP, and LFABP were significantly higher in cases [25.6 (.7), 10.8 (.5), and 47.8 (1.9)] than controls [19.1 (.8), 7.7 (2), 22. 9 (.5)], P &lt; .001. All FABPs were significantly associated with cirrhosis, higher Child Pugh Score (CTP), advanced stage in Barcelona clinic liver cancer stage (BCLC), higher AFP levels, vascular invasion and thrombosis, and tumor nodularity. Median OS (months) (95%CI) were significantly short in patients with higher level of AFABP, HFABP, and LFABP [9.3 (6.8-11.9), 9.4 (6.8-11.9), and 11.1 (8.8-13.3)] as compared to patients with low levels [16.4 (13.8-18.9), 16.4 (14.2-18.6), and 17.9 (14.9-20.9) respectively (P &lt; .01). The significance was observed in non-viral related HCC for LFABP and HFABP, but not AFBABP. Conclusions: To the best of our knowledge, we describe the largest study correlating FABPs levels with clinical and prognostic characteristics of HCC. Higher levels were associated with poor survival. These findings suggest that LFABP and HFABP may be used as potential prognostic biomarkers for non-viral-related HCC.

Hypo-phosphorylated CD147 promotes migration and invasion of hepatocellular carcinoma cells and predicts a poor prognosis

PurposeCD147 is a tumor-associated antigen that plays a key regulatory role in tumor invasion and distant metastasis. However, the exact role of CD147 phosphorylation, which is deregulated during cancer progression, is unknown. Here, the effects of CD147 phosphorylation on the malignant behavior of hepatocellular carcinoma (HCC) cells and its possible underlying mechanisms are explored.MethodsAn in situ Duolink-proximity ligation assay (PLA) was used to detect CD147 phosphorylation. Tandem mass spectrometry was employed to identify the phosphorylation sites of CD147. The effects of CD147 phosphorylation on the malignant behavior of HCC cells were evaluated using scratch wound healing assays, transwell invasion assays and cell cycle assays. The genes regulated by CD147 phosphorylation were detected by RNA sequencing.ResultsWe identified phosphorylated serine-246 in the C terminus of CD147 in primary HCC tissues, whereas serine to alanine substitution mutation analysis suggested that CD147 is phosphorylated mainly at serine-252 in HCC-derived Huh-7 cells. Recovery expression of S246A/S252A mutants in CD147 knockout cells revealed significantly increased migration and invasion capacities compared to wildtype CD147 expressing cells. Cyclophilin A (CyPA) treatment decreased the phosphorylation level of CD147, whereas NIMA-related kinase 6 (NEK6) increased the CD147 phosphorylation level. Moreover, the CD147 phosphorylation level was found to be dramatically decreased in HCC tissues in patients with distant metastases, and a low phosphorylation level of CD147 was found to be associated with a high serum AFP level, recurrence and a poor overall survival.ConclusionsFrom our data we conclude that hypo-phosphorylated CD147 promotes the migration and invasion of HCC cells and correlates with an unfavorable prognosis in HCC patients, indicating that targeting the aberrantly hypo-phosphorylated form of CD147 may be instrumental for the development of novel therapeutic modalities directed against HCC metastasis.

Retraction: The Human Homolog of Drosophila Headcase Acts as a Tumor Suppressor through Its Blocking Effect on the Cell Cycle in Hepatocellular Carcinoma.

The molecular pathogenesis of hepatocellular carcinoma (HCC) is heterogeneous and extremely complex. Thus, for individual molecular targeted therapy, novel molecular markers are needed. The abnormal expression of the human homolog of Drosophila headcase (HECA homo) has been found in pancreatic, colorectal, and oral squamous cell carcinoma. Studies of oral squamous cell carcinoma have also demonstrated that the HECA homo protein can be negatively controlled by the Wnt-pathway and transcription factor 4 (TCF4) and can slow cell division by interacting with cyclins and CDKs. However, the role of HECA in HCC has not been reported elsewhere. Here, immunohistochemical analysis revealed that the downregulation of HECA homo protein occurred in 71.0% (66/93) of HCC cases and was positively correlated with a poorly differentiated grade, high serum AFP level, liver cirrhosis and large tumor size. The expression of HECA homo was detected in five live cell lines. In vitro, the overexpression of HECA homo in HepG2, Huh-7 and MHCC-97H cells could inhibit cell proliferation and colony formation and induce G1 phase arrest. In contrast, the downregulation of HECA homo could promote cell proliferation, colony formation and the cell cycle process. However, neither the overexpression nor downregulation of HECA homo in the three cell lines could affect cell migration or invasion. Collectively, HECA homo is regularly expressed in normal live cells, and the HECA homo protein level is heterogeneously altered in HCC, but the downregulation of HECA homo is more common and positively correlated with several malignant phenotypes. The HECA homo protein can slow cell proliferation to some extent primarily through its blocking effect on the cell cycle. Hence, the HECA homo protein may act as a tumor suppressor in HCC and might be a potential molecular marker for diagnostic classification and targeted therapy in HCC.

Predictors of hepatocellular carcinoma recurrence associated with the use of direct-acting antiviral agent therapy for hepatitis C virus after curative treatment: A prospective multicenter cohort study.

BackgroundPrevious studies have suggested an association between the use of direct‐acting antiviral agents (DAAs) for treating hepatitis C virus (HCV) infection and the resulting decrease in the incidence of hepatocellular carcinoma (HCC); however, it is unclear whether DAAs prevent the recurrence of HCC after curative treatment for HCC. This study aimed to prospectively investigate HCC recurrence and its predictors after curative treatment for HCC.MethodsA total of 3012 patients with chronic HCV infection, with or without cirrhosis, who were treated with DAAs were enrolled between January 1, 2015 and January 31, 2017 as per the institutional review board approved study protocol at 15 institutions, including 10 university hospitals and five high‐volume centers in the Kyusyu area of Japan. Of the 3012 patients, 459 patients who had HCC but were cured with surgery or ablation therapy (curative treatment) before the use of DAAs were included in the analysis.ResultsDuring a mean follow‐up period of 29.4 months, 217 (47.2%) patients developed HCC recurrence. The median time to recurrence was 34.0 months, and the 1‐, 2‐, and 3‐year cumulative HCC recurrence rates were 27.1%, 43.4%, and 50.8%, respectively. The risk factors for HCC recurrence were the α‐fetoprotein (AFP) level before DAA therapy (P = 0.0047) and the number of curative treatments for HCC before DAA therapy (P < 0.0001).ConclusionsA high AFP level and multiple occurrences of HCC before DAA therapy are associated with a high risk for HCC recurrence after curative treatment. Follow‐up after DAA therapy should include special attention to the abovementioned risk factors.

Second Trimester Serum Biomarker Screen for Fetal Aneuploidies as a Predictor of Preterm Delivery: A Population-Based Study

Objective: To determine the association between second-trimester serum Down syndrome screening (alpha-fetoprotein [AFP] free beta-human chorionic gonadotropin [b-hCG] unconjugated estriol [uE3]) and preterm birth and to create predictive models for preterm birth. Methods: Secondary analysis on a prospective database of pregnancies undergoing second-trimester screen with complete follow-up. The multiples of medians (MoM) of the biomarkers were compared between the group of term, preterm (< 37 weeks), early preterm (< 34 weeks), and very early preterm (< 32 weeks) delivery. Predictive models were developed based on adjusted MoMs and logistic regression and diagnostic performances in predicting preterm birth were determined. Results: Of 20,780 pregnancies, 1,554 (7.5), 363 (1.7), and 158 (0.8%) had preterm, early preterm, and very early preterm birth respectively. High levels of AFP and b-hCG but low levels of uE3 were significantly associated with higher rates of preterm, early preterm and very early preterm delivery. The predictive models had diagnostic performance in predicting preterm birth with the areas under the ROC curve of 0.688, 0.534, 0.599, and 0.718 for AFP, b-hCG, uE3, and combined biomarkers respectively. Conclusion: The second trimester Down syndrome screening could also be used as a tool of risk identification of preterm birth in the same test, without extra-effort and extra-cost.

Pattern of hepatocellular carcinoma recurrence following living donor liver transplantation

Background: The aim of the present study was to identify the pattern of HCC recurrence after LDLT for early detection and management Methods: From April 2003 to October 2014, the record of 60 patients who underwent LDLT for HCC at the National Liver Institute, Menoufia University, Egypt, were retrospectively reviewed. Results: Seven patients (11.7 per cent) had HCC recurrence after LDLT. Pretransplant α-fetoprotein (AFP) > 1000 ng/mL, tumour grade and microvascular invasion were the incriminated risk factors for recurrence. Three patients (42.8 per cent) had intrahepatic and extrahepatic recurrence (lung and bone), two patients (28.6 per cent) had only extrahepatic recurrence in bones and two patients (28.6 per cent) had only intrahepatic recurrence. Management was as follows: two patients (28.6 per cent) had surgical excision of intrahepatic recurrence and extrahepatic metastasis, two patients (28.6 per cent) underwent radiotherapy for bone metastasis, one patient (14.2 per cent) underwent intraoperative radiofrequency ablation for liver recurrence and two patients (28.6 per cent) received sorafenib as medical treatment. The mean time of recurrence was 19.7 months, and mean survival was 29 months. Conclusion: The majority of HCC recurrences after LDLT are extrahepatic and occur mainly in the first 2 years; strict follow up is required during this period. A high level of pretransplant serum AFP and microvascular invasion are risk factors for tumour recurrence and should be taken into account when selecting candidates for LDLT.

Immunohistochemical Expression of Epidermal Growth Factor Receptor in Hepatocellular Carcinoma.

Epidermal growth factor receptor (EGFR) signaling plays an important role in various cancers, including hepatocellular carcinoma (HCC). We aimed to evaluate immunoexpression of EGFR in HCC and surrounding non-tumor liver tissue and to correlate it to multiple clinicopathologic data. We analyzed 60 patients with HCC for multiple clinicopathologic characteristics and survival. Presence of the immunosignal and the percentage of positive tumor cells at the whole tumor tissue sample and adjacent cirrhotic liver tissue were semi-quantitatively determined. Nineteen patients (31.67%) were female and 41 (68.33%) were male ranging in age from 31 to 85 years, median 61.88±10.51. Mean survival time for female patients was 8.86±1.76 months, for male 13.03±1.50 months and overall survival was 11.6051±1.19 months. The most patients had: T2 status (41.67%), no enlarged lymph nodes (90%), vascular invasion (63.33%) and well differentiated (43.33%) tumors. EGFR immunoexpression was determined in range from 0% to 100% in both tumor and non-tumor tissue with mean value of 39.58% in tumor and 86.86% in cirrhotic tissue (p<0.00). Higher percent of tumor EGFR positive cells were found in cases with higher T status, higher levels of AFP and poorly differentiated carcinoma, but not significantly. Lower percent of tumor EGFR positive cells were found in patients with vascular invasion and enlarged lymph nodes, but also not significantly. EGFR expression in tumor tissue significantly influenced survival of the patients (p<0.05). The study showed that expression of EGFR in lower percentage of tumor cells was associated to favorable prognosis, making it a potential prognostic marker and therapeutic target.

FokI polymorphism in vitamin D receptor gene and its association with hepatocellular carcinoma in Egyptian patients with chronic liver disease

AimTo assess the relationship between vitamin D receptor (VDR) gene polymorphisms at FokI site and hepatocellular carcinoma (HCC) in chronic liver disease patients. MethodsThe study enrolled 70 patients with chronic liver disease; of them 20 patients had HCC. Specific primer sequences of the VDR FokI C > T single nucleotide polymorphisms rs10735810, was used to amplify VDR gene. Allele specific restriction enzymes FokI was used to digest the amplified PCR products and the bands of specific PCR-RFLP fragments were resolved on 2% Ethidium bromide agrose gel electrophoresis. FokI genotypes was identified by their specific banding pattern on the gel electrophoresis via ultraviolet illumination. ResultsIn chronic liver disease the frequency of FF, Ff and ff genotypes were 50%, 43% and 7% respectively. The wild type F and the mutant f allele had a frequency of 71%, 29% respectively. In HCC group the frequencies of FokI FF, Ff and ff genotypes were 55%, 30% and 15%; relative to 48%, 48% and 4% in non HCC respectively. F allele had a frequency of 72%, and 70%; and f allele had 28% and 30% respectively. VDR FokI genotypes and allele frequencies were not significantly associated with HCC in chronic liver diseases. The carriage of FokI polymorphisms ff genotype was associated with increased tumor size and higher AFP levels. FokI polymorphisms did not show any significance association with tumor stage, portal vein invasion or lymph node metastasis. ConclusionVDR FokI genotypes and allele frequencies were neither significantly associated with HCC, HCV nor were risk for progression of chronic liver disease into decompensated liver disease.

Restoration of FBP1 suppressed Snail-induced epithelial to mesenchymal transition in hepatocellular carcinoma

Fructose-1,6-bisphosphatase (FBP1), one of the rate-limiting gluconeogenic enzymes, plays critical roles in several cancers and is treated as a tumour suppressor. However, its role in hepatocellular carcinoma (HCC) is unclear. Here, we demonstrated that FBP1 was significantly inhibited during Snail-induced epithelial to mesenchymal transition (EMT) and tissues in HCC. Restoration of FBP1 expression in HCC cancer cells suppressed EMT phenotype, tumour migration and tumour growth induced by Snail overexpression in SMMC-7721 cells. Gene set enrichment analyses revealed significantly enriched terms, including WNT, Notch, ESC, CSR and PDGF, in the group with high Snail and low FBP1 compared with those with low Snail and high FBP1. Low FBP1 expression was significantly correlated with higher AFP level, satellite nodules, portal vein tumour thrombus, and advanced tumour stage. Survival analyses showed that FBP1 was an independent prognostic factor for overall survival and recurrence-free survival. In conclusion, our study revealed a vital role for FBP1 in Snail-induced EMT and prognostic prediction in HCC.

A case of giant panda ovarian cancer diagnosis and histopathology

BackgroundOvarian cancer is diagnosed clinically by detecting ovarian cancer-related factors and markers. Here, we report a case of giant panda ovarian tumor metastasis with a combination of clinical and histopathological diagnosis.Case presentationHistopathological studies revealed severe lesions and tumor cells in the ovaries, lungs, spleen, kidneys and perianal tissue. Immunohistochemistry staining showed that the ovarian cancer markers B7-H4, CA125, and HE4 were highly expressed in the lungs, kidneys, spleen, ovaries and perianal tissue. Tumor marker tests detected significantly high levels of AFP in serum.ConclusionClinical biomarkers combined with histopathology can provide a more accurate diagnosis of ovarian cancer metastasis and identification of ovarian cancer types than either method alone. The giant panda’s death may be due to granulosa cell tumor and theca cell tumor metastasis causing multiple organ dysfunction or even failure.

Impact of DAA-Based Therapy on Alfa-Fetoprotein Level in Chronic Hepatitis C Patients

Introduction: Backgrounds: Chronic hepatitis C virus (HCV) infection is a leading cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma worldwide. High AFP level has been identified as a risk factor for the development of HCC in chronic hepatitis C patients. The aim of this study is to investigate the impact of DAA-based triple therapy on serum AFP levels of patients with chronic hepatitis C. Methods: We reviewed records of 132 consecutive patients with chronic hepatitis C who received DAAbased therapy over past 16 months. Inclusion criteria: patients who completed treatment, had baseline AFP within 3 months of treatment, and AFP within 6 months post-treatment. 54 patients were excluded (52: no AFP level, and 2 patient were lost to follow). Results: 78 patients completed the treatment: age: 24-78 year, 45 males, 33 females. Of these patients: 46 genotype 1, 22 genotype 2, 10 genotypes 3 and 4. All had viral loads, and fibrosis score. 76 patients achieved SVR at 3 months and 6 months pots-treatment. 2 patients relapsed. Conclusion: Discussion: AFP is commonly used for surveillance for HCC. In this review, there was universal and lasting ( beyond 6 months posttreatment) reduction in AFP below 50% of baseline level in all patients s who received treatment even in those who relapsed, and across all genotypes, most pronounced in patients with markedly high pretreatment level and those with documented HCC. This phenomenon supports the evidence that DAA therapy may prevent HCC in hepatitis C patients. Conclusion: DAA therapy of hepatitis C-infected patient effectively reduce the level of AFP and so potentially may prevent HCC in this group of patients2851_A Figure 1. AFP: Pre- and posttreatment

Identification of WDR12 as a novel oncogene involved in hepatocellular carcinoma propagation.

BackgroundHepatocellular carcinoma (HCC) is one of the most common malignant cancer worldwide. Importantly, the precise mechanisms causing HCC pathogenicity are still unknown. The identification of potential oncogenes plays significant roles in finding novel therapeutic targets for human HCC.PurposeWDR12 (WD repeat protein 12), a member of WD repeats family, plays crucial roles in the ribosome biogenesis pathway. However, Whether WDR12 contributes to HCC development remains unknown. The objective of this study was to elucidate the role of WDR12 in HCC development.MethodsThe expression level of WDR12 in HCC tissues and adjacent non-tumor tissues were detected form Gene Expression Omnibus (GEO) database. The expression level of WDR12 in HCC cell lines were examined by RT-PCR and western blot. Kaplan-Meier analysis were used to analyze the effect of WDR12 level on overall and disease-free survival of HCC patients. To examine whether WDR12 supports development of HCC, we inhibited expression of WDR12 by using an shRNA-encoding lentivirus system. Effects of WDR12 knockdown were evaluated on cell-growth, cell-proliferation and cell-migration. The mechanisms involved in HCC cells growth, proliferation and migration were analyzed by western blot assay.ResultsIn silico analysis of HCC data sets showed that elevated expression of WDR12 correlated with high serum AFP level, high vascular invasion, high histologic grade and high TNM stage in HCC patients. Furthermore, up-regulated expression of WDR12 significantly correlated with the short overall survival and recurrence time of HCC patients. The shRNA-mediated knockdown of WDR12 expression resulted in reduced proliferation and migration of HepG2 and Huh-7 cells. Notably, inhibition of WDR12 resulted in decreased phosphorylation of AKT, mTOR and S6K1.ConclusionOur study indicates that WDR12 contributes to HCC propagation, and indicates that suppression of WDR12 may be a potential strategy for human HCC treatment.

NFATc1 is a tumor suppressor in hepatocellular carcinoma and induces tumor cell apoptosis by activating the FasL-mediated extrinsic signaling pathway.

Nuclear factor of activated T cells (NFAT) is a family of transcription factors that have important functions in many tumors. However, the expression level and functional role of NFAT in hepatocellular carcinoma (HCC) remain unclear. In this study, we showed that NFATc1 expression was decreased in both HCC tissues and cell lines. Low expression of NFATc1 was correlated with larger tumor size, advanced tumor‐node‐metastasis (TNM) stage, high serum AFP level, and liver cirrhosis. Furthermore, patients with low NFATc1 expression exhibited poor prognosis. Ectopic expression of NFATc1 in HCC cells inhibited proliferation and colony formation, leading to G1 arrest and induction of apoptosis. In addition, we demonstrated that NFATc1 increased Fas ligand (FasL) expression by directly binding to its promoter and activated the extrinsic apoptotic pathway. NFATc1 and FasL expression patterns and their prognostic value for patients with HCC were also evaluated in TCGA Liver Hepatocellular Carcinoma database. Knock‐down of FasL expression by siRNA in HCC cell lines abolished NFATc1's antiproliferative and pro‐apoptotic effects. In conclusion, NFATc1 is frequently inactivated in HCC and functions as a tumor suppressor in liver carcinogenesis. Ectopic expression of NFATc1 in HCC cells induces apoptosis by activating the FasL‐mediated extrinsic signaling pathway.