Identification of MCM4 as a Prognostic Marker of Hepatocellular Carcinoma

Object. Hepatocellular carcinoma is one of the most common malignant tumors worldwide owing to its complicated molecular and cellular heterogeneity and its high incidence rate every year. It is an urgent need to search for new efficient molecular markers of HCC to reduce mortality and improve HCC prognosis. In this article, MCM4, a member of a family of proteins closely related to DNA replication and cell proliferation, was selected as a potential biomarker of HCC prognosis. Methods. MCM4 expression difference in HCC were analyzed from TCGA and GEO data and verified by real-time PCR and western blot. ROC curve was used to analyze the diagnostic value of MCM4 and AFP. Additionally, the relationship between MCM4 and stage or nodal metastasis status or grade or age in TCGA cohort with HCC was observed from the UALCAN website. The univariate and multivariate Cox and functional analyses were done to explore the prognostic value of MCM4 in TCGA cohort. Results. It was found that MCM4 was significantly highly expressed in HCC tissues from TCGA, GEO, and experimental data. Furthermore, ROC curve analysis showed that MCM4 was superior to be a diagnostic biomarker than AFP from TCGA ( AU C MCM 4 = 0.9461 , AU C AFP = 0.7056 ) and GEO (GSE19665: AU C MCM 4 = 0.8800 , AU C AFP = 0.5100 ; GSE64041 AU C MCM 4 = 0.8038 , AU C AFP = 0.6304 ). AUC of MCM4 from real-time PCR result in 60 pairs of HCC and adjacent tissues was 0.7172, demonstrating the prediction value of MCM4. Besides, different expression tendencies of MCM4 among different stages or nodal metastasis status or grade or age were observed from the UALCAN website. In addition, multiROC analysis showed the advantage of MCM4 as a survival prediction at 1, 3, and 5 years with the higher AUC at 0.69 of 1 year, 0.65 of 3 years, and 0.61 of 5 years. It was shown that MCM4 was independently associated with OS in univariate and multivariate Cox analysis. And GSEA displayed that MCM4 was highly enriched in KEGG_CELL_CYCLE signaling pathway following higher correlation positively with CDC6, PLK1, CRC1, and BUB1B in HCC. Conclusion. MCM4 might be a potential biomarker in guiding the prognostic status of HCC patients.

Download Full-text

CircRNA profiling identifies circRNF180 as a tumor suppressor in hepatocellular carcinoma

Epigenomics ◽

10.2217/epi-2020-0385 ◽

2021 ◽

Vol 13 (7) ◽

pp. 513-530

Author(s):

Xi Zeng ◽

Chao Tan ◽

Meile Mo ◽

Xiaoling Qin ◽

Xiaoyun Ma ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Tumor Suppressor ◽

Real Time ◽

Real Time Pcr ◽

Expression Profiles ◽

Cell Counting ◽

Migration And Invasion ◽

Quantitative Real Time Pcr ◽

Potential Biomarker ◽

Hcc Cells

Aim: To explore the expression profiles and functions of circRNAs in hepatocellular carcinoma (HCC). Materials & methods: We obtained circRNA expression profiles through RNA sequencing. Expression levels of circRNAs were confirmed by quantitative real-time PCR. The effects on HCC progression were determined using Cell Counting Kit 8, clone formation and transwell assays. Results: We identified 114 upregulated and 144 downregulated circRNAs in HCC tissues. The results of quantitative real-time PCR showed that circGNAO1, circRNF180 and circMERTK were significantly downregulated in HCC tissues, whereas circSNX6 was significantly upregulated. CircRNF180 was associated with microvascular invasion. Overexpression of circRNF180 inhibits the proliferation, colony formation, migration and invasion of HCC cells. Conclusion: CircRNF180 may function as a tumor suppressor and could serve as a potential biomarker and therapeutic target in HCC.

Download Full-text

The effect of BKV reactivation on cytokines behavior in kidney transplanted patients

BMC Nephrology ◽

10.1186/s12882-021-02645-y ◽

2022 ◽

Vol 23 (1) ◽

Author(s):

Zahra Rahimi ◽

Ramin Yaghobi ◽

Afsoon Afshari ◽

Jamshid Roozbeh ◽

Mohammad Javad Mokhtari ◽

...

Keyword(s):

Real Time ◽

Roc Curve ◽

Real Time Pcr ◽

Curve Analysis ◽

Expression Level ◽

Control Group ◽

Roc Curve Analysis ◽

Tnf Α ◽

Ifn Γ ◽

Inactive Group

Abstract Background BK virus associated nephropathy (BKVAN) is one of the common causes of graft loss among kidney transplanted recipients (KTRs). The current treatment for BKV nephropathy is decreasing the immunosuppressive regimen in KTRs. Interleukin-27 (IL-27) is a multifunctional cytokine that might be the front-runner of an important pathway in this regard. Therefore, in current study it is tried to evaluate the changes in the expression level of IL-27 and some related molecules, resulting from BKV reactivation in KTR patients. Methods EDTA-treated blood samples were collected from all participants. Patients were divided into two groups, 31 kidney transplant recipients with active and 32 inactive BKV infection, after being monitored by Real time PCR (Taq-Man) in plasma. Total of 30 normal individuals were considered as healthy control group. Real time PCR (SYBR Green) technique is used to determine the expression level of studied genes. Results The results of gene expression comparisons showed that the expression level of IL-27, IFN-γ, TNF-α, TNFR2 and IRF7 genes was significantly higher in inactive group in comparison to active group. The expression level of TLR4 was lower in both active and inactive groups in comparison to control group. ROC curve analysis showed that IL-27 and IRF7 are significantly different amongst other studied genes. Finally, the analyses revealed that the expression level of most of the studied genes (except for TNF-α and TLR4) have significant correlation with viral load. Conclusions Our findings revealed that IL-27, IFN-γ, TNF-α, TNFR2 and IRF7 expression level is higher in inactive group and TLR4 expression level is lower in patients’ groups in comparison to control group. Also, ROC curve analysis showed IL-27 and IRF7 can significantly differentiate studied groups (BKV active vs. inactive). Therefore, these results might help elucidating the pattern in charge of BKV reactivation in kidney transplanted patients.

Download Full-text

Implication of efflux pumps and ERG11 genes in resistance of clinical Trichosporon asahii isolates to fluconazole

Journal of Medical Microbiology ◽

10.1099/jmm.0.001236 ◽

2021 ◽

Vol 70 (3) ◽

Author(s):

S. Abbes ◽

H. Sellami ◽

S. Neji ◽

H. Trabelsi ◽

F. Makni ◽

...

Keyword(s):

Real Time ◽

Roc Curve ◽

Real Time Pcr ◽

Statistical Study ◽

Resistance Mechanism ◽

Resistance Mechanisms ◽

Curve Analysis ◽

Roc Curve Analysis ◽

Trichosporon Asahii ◽

Over Expression

Introduction. Trichosporon asahii has been recognized as an opportunistic agent having a limited sensitivity to antifungal treatment. Hypothesis/Gap Statement. Molecular mechanisms of azole resistance have been rarely reported for Trichosproron asahii. Similar to other fungi, we hypothesized that both ERG11 gene mutation and efflux pumps genes hyper-expression were implicated. Aim. The current work aimed to study the sensitivity of clinical T. asahii isolates to different antifungal agents and to explore their resistance mechanisms by molecular methods including real-time PCR and gene sequencing. Methods. The sensitivity of T. asahii isolates to fluconazole, amphotericin B and voriconazole was estimated by the Etest method. Real-time PCR was used to measure the relative expression of Pdr11, Mdr and ERG11 genes via the ACT1 housekeeping gene. Three pairs of primers were also chosen to sequence the ERG11 gene. This exploration was followed by statistical study including the receiver operating characteristic (ROC) curve analysis to identify a relationship between gene mean expression and the sensitivity of isolates. Results. In 31 clinical isolates, the resistance frequencies were 87, 16.1 and 3.2 %, respectively, for amphotericin B, fluconazole and voriconazole. Quantitative real-time PCR demonstrated that only Mdr over-expression was significantly associated with FCZ resistance confirmed by univariate statistical study and the ROC curve analysis (P <0.05). The ERG11 sequencing revealed two mutations H380G and S381A in TN325U11 (MIC FCZ=8 µg ml−1) and H437R in TN114U09 (MIC FCZ=256 µg ml−1) in highly conserved regions (close to the haem-binding domain) but their involvement in the resistance mechanism has not yet been assigned. Conclusion. T. asahii FCZ resistance mechanisms are proven to be much more complex and gene alteration sequence and/or expression can be involved. Only Mdr gene over-expression was significantly associated with FCZ resistance and no good correlation was observed between FCZ and VCZ MIC values and relative gene expression. ERG11 sequence alteration seems to play a major role in T. asahii FCZ resistance mechanism but their involvement needs further confirmation.

Download Full-text

Utility of Alpha-Fetoprotein-L3 and Golgi Protein 73 for Diagnosis of Hepatocellular Carcinoma

10.51429/ejmm29402 ◽

2020 ◽

Vol EJMM29 (4) ◽

pp. 9-15

Author(s):

Rania A. El-Kady ◽

Mohammed M. El-Naggar ◽

ehab A. Abd El-Shakour ◽

Monir H. Bahgat

Keyword(s):

Hepatocellular Carcinoma ◽

Roc Curve ◽

Malignant Tumors ◽

Curve Analysis ◽

Alpha Fetoprotein ◽

Enzyme Linked Immunosorbent Assay ◽

Roc Curve Analysis ◽

Median Serum ◽

Significant Difference ◽

Golgi Protein

Background: Hepatocellular carcinoma (HCC) is amongst the most common malignant tumors that carries a poor prognosis. Clinically, alpha-fetoprotein (AFP) is the most extensively used serum biomarker for diagnosing HCC. Objectives: The current study was conducted to explore the diagnostic value of serum levels of alpha-fetoprotein-L3 (AFP-L3) and Golgi protein 73 (GP73) regarding HCC, and to determine the diagnostic accuracy of these biomarkers when used individually as well as in combination with AFP. Methodology: Blood samples were collected from 50 patients with HCV-related cirrhosis (25 subjects with HCC and 25 without HCC) recruited from the outpatient clinics of the Specialized Internal Medicine Hospital, Mansoura University, Egypt. Serum concentrations of AFP-L3 and GP73 were evaluated using enzyme-linked immunosorbent assay (ELISA). Diagnostic performance of AFP-L3 and GP73 was determined by receiver operating characteristic (ROC) curve analysis. Results: Overall, the median serum level of AFP-L3 was higher in the HCC group compared to the cirrhotic group (p=0.05). Moreover, a statistically-significant difference was observed between the median serum value of GP73 in HCC patients compared to those with cirrhosis (p < 0.001). The ROC curve analysis showed that the area under the ROC curve (AUROC) values for AFP, AFP-L3 and GP73 were 0.88, 0.67 and 0.83, respectively. Of the 3 biomarkers, GP73 demonstrated the highest sensitivity (88%). The AUROC for AFP and AFP-L3 combination was 0.85, whereas that for AFP and GP73 was 0.90. Conclusion: Our findings indicate that GP73 is more sensitive than AFP and AFP-L3 in diagnosing HCC. Furthermore, the combined determination of GP73 and AFP could improve the diagnostic ability of HCC.

Download Full-text

Baseline apparent diffusion coefficients: Validation study of new predictor of survival in patients with unresectable hepatocellular carcinoma following chemoembolization

Journal of X-Ray Science and Technology ◽

10.3233/xst-200827 ◽

2021 ◽

pp. 1-10

Author(s):

Lichao Xu ◽

Shiqin Wang ◽

Shengping Wang ◽

Ying Wang ◽

Wentao Li ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Roc Curve ◽

Cox Regression ◽

Response To Treatment ◽

Roc Curve Analysis ◽

Adc Value ◽

Apparent Diffusion Coefficients ◽

Apparent Diffusion ◽

Response Biomarker ◽

The Difference

OBJECTIVES: To investigate whether the baseline apparent diffusion coefficient (ADC) can predict survival in the hepatocellular carcinoma (HCC) patients receiving chemoembolization. MATERIALS AND METHODS: Diffusion-weighted MR imaging of HCC patients is performed within 2 weeks before chemoembolization. The ADC of the largest index lesion is recorded. Responses are assessed by mRECIST after the start of the second course of chemoembolization. Receiver operating characteristic (ROC) curve analysis is performed to evaluate the diagnostic performance and determine optimal cut-off values. Cox regression and Kaplan–Meier survival analyses are used to explore the differences in overall survival (OS) between the responders and non-responders. RESULTS: The difference is statistically significant in the baseline ADC between the responders and non-responders (P < 0.001). ROC analyses indicate that the baseline ADC value is a good predictor of response to treatment with an area under the ROC curve (AUC) of 0.744 and the optimal cut-off value of 1.22×10–3 mm2/s. The Cox regression model shows that the baseline ADC is an independent predictor of OS, with a 57.2% reduction in risk. CONCLUSION: An optimal baseline ADC value is a functional imaging response biomarker that has higher discriminatory power to predict tumor response and prolonged survival following chemoembolization in HCC patients.

Download Full-text

Clinical significance of fucosylated GP73 in the differential diagnosis of hepatocellular carcinoma

The International Journal of Biological Markers ◽

10.1177/1724600818796646 ◽

2018 ◽

Vol 33 (4) ◽

pp. 439-446 ◽

Cited By ~ 5

Author(s):

Yunsheng Zhao ◽

Lina Zhang ◽

Lijing Huo ◽

Liu Pei ◽

Qiuping Li ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Differential Diagnosis ◽

Roc Curve ◽

Enzyme Linked Immunosorbent Assay ◽

Diagnostic Efficiency ◽

Roc Curve Analysis ◽

Diagnostic Significance ◽

Node Metastasis ◽

Tumor Node Metastasis Stage ◽

Normal Controls

Objective: To investigate the clinical value of fucosylated GP73 (Fuc-GP73) levels for differential diagnosis of hepatocellular carcinoma from other liver diseases. Methods: Serum specimens were collected from 50 patients with hepatocellular carcinoma, 60 patients with other digestive system diseases (ODSD), and 40 normal controls. Lectin affinity chromatography column combining with the enzyme-linked immunosorbent assay (ELISA) using the ELISA index was utilized to measure the level of Fuc-GP73. By receiver operating characteristic (ROC) curve analysis its sensitivity and specificity were used to evaluate the diagnostic significance of Fuc-GP73 in hepatocellular carcinoma. Results: The median serum Fuc-GP73 level of hepatocellular carcinoma (20.4 μg/L) was much higher than that of ODSD patients (1.8 μg/L) and the normal controls group (0.3 μg/L), significantly ( P <0.01). There was no significant correlation between serum Fuc-GP73 level and sex, age, and tumor size in the hepatocellular carcinoma group ( P > 0.05); however, it was related to tumor, node, metastasis stage and lymph node metastasis ( P <0.05). The area under the ROC curve (AUC) of Fuc-GP73 to detect hepatocellular carcinoma alone was 0.885; with the prespecified specificity of 95%, the sensitivity and the cutoff value were 82% and 3.1 μg/L. In addition, the combined test of Fuc-GP73 with other biomarkers can improve the clinical diagnostic efficiency; the AUC can reach to 0.983; and with the prespecified specificity of 95% its sensitivity increased to 94%. Conclusion: Fuc-GP73 can act as a superior glycobiomarker for the differential diagnosis of hepatocellular carcinoma; its combined detection with other biomarkers can improve diagnostic accuracy.

Download Full-text

TaqMan real-time PCR assay for specific detection of Opisthorchis viverrini DNA in Thai patients with hepatocellular carcinoma and cholangiocarcinoma

Experimental Parasitology ◽

10.1016/j.exppara.2008.01.018 ◽

2008 ◽

Vol 119 (2) ◽

pp. 217-224 ◽

Cited By ~ 14

Author(s):

Nithikoon Suksumek ◽

Kawin Leelawat ◽

Surang Leelawat ◽

Bruce Russell ◽

Usa Lek-Uthai

Keyword(s):

Hepatocellular Carcinoma ◽

Real Time ◽

Real Time Pcr ◽

Opisthorchis Viverrini ◽

Specific Detection ◽

Pcr Assay

Download Full-text

Suitable reference genes for real-time PCR in human HBV-related hepatocellular carcinoma with different clinical prognoses

BMC Cancer ◽

10.1186/1471-2407-9-49 ◽

2009 ◽

Vol 9 (1) ◽

Cited By ~ 41

Author(s):

Li-Yun Fu ◽

Hu-Liang Jia ◽

Qiong-Zhu Dong ◽

Jin-Cai Wu ◽

Yue Zhao ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Real Time ◽

Real Time Pcr ◽

Reference Genes ◽

Suitable Reference

Download Full-text

Serum Zonulin in HBV-Associated Chronic Hepatitis, Liver Cirrhosis, and Hepatocellular Carcinoma

Disease Markers ◽

10.1155/2019/5945721 ◽

2019 ◽

Vol 2019 ◽

pp. 1-6 ◽

Cited By ~ 2

Author(s):

Xin Wang ◽

Ming-ming Li ◽

Ye Niu ◽

Xin Zhang ◽

Ji-bin Yin ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Liver Cirrhosis ◽

Chronic Hepatitis ◽

Diagnostic Accuracy ◽

Roc Curve ◽

Healthy Subjects ◽

Serum Levels ◽

Curve Analysis ◽

Advanced Stage ◽

Roc Curve Analysis

Background. The gut microbiota is involved in the occurrence and development of chronic liver diseases. Zonulin is considered a marker of intestinal permeability. The purpose of this study was to assess zonulin levels in patients with chronic hepatitis B (CHB), HBV-associated liver cirrhosis (LC), and HBV-associated hepatocellular carcinoma (HCC). Materials and Methods. The study population consisted of 90 HBV-associated HCC patients, 90 HBV-associated LC patients, 90 CHB patients, and 90 healthy subjects. Serum levels of zonulin and AFP were determined. The diagnostic accuracy of each marker was evaluated using receiver operating characteristic (ROC) curve analysis (AUC). Results. Serum zonulin levels were significantly higher in patients with HCC than in patients with LC or CHB or healthy subjects (p<0.001). Moreover, the zonulin levels were increased in the advanced stage of LC and HCC. ROC curve analysis revealed that serum zonulin could be used to differentiate CHB from cirrhosis. In addition, the combination of zonulin and AFP exhibited a significantly larger AUC compared with zonulin or AFP alone. Conclusions. Serum zonulin levels were significantly increased both in LC and in HCC and correlated with the advanced stage of LC and HCC. Moreover, the combination of zonulin and AFP confers significant benefit to diagnostic accuracy in differentiating LC from HCC.

Download Full-text