Evaluation of TGFβ, XPO4, elF5A2 and ANGPTL4 as biomarkers in HCC.
ABSTRACT: Hepatocellular carcinoma (HCC) is the most common type of liver cancer, and the fourth leading cause of cancer mortality worldwide. It is often diagnosed at an advanced stage, and hence typically has a poor prognosis. A number of distinct molecules have been recently identified as playing a role in the control of cancer progression. However, patients with HCC have a highly variable clinical course, indicating that HCC comprises several biologically distinctive subgroups reflecting a molecular heterogeneity of the tumors. To evaluate potential biomarkers in HCC, we employed multiple methods in this study, including qPCR, immunostaining methods and tissue microarrays (TMAs), as well as histological and pathological analysis, to assess TGFβ, XPO4, elF5A2 and ANGPTL4 in cancerous and paracancerous liver tissues from 280 patients suffering from liver cancer. Our results found that all four indicators were located in the cytoplasm and distributed in cancerous and paracancerous liver tissues. Generally, there were higher levels of these indicators in paracancerous, compared with cancerous, liver tissues. These four indicators were correlated and modulated among each other. In connection with patient clinical and revisit information, statistical analysis determined that TGFβ1 in paracancerous liver tissue was positively correlated with tumor size. Higher production of TGFβ1 in paracancerous liver tissue was always associated with bigger liver tumors. XPO4 in cancerous liver tissue and TGFβ1 in paracancerous liver tissue were positively correlated with tumor differentiation. TGFβ1, ANGPTL4 and elF5A2 were also positively correlated with the T classification of tumors. Additionally, higher levels of XPO4 in cancerous liver tissue suggested that the patient would have a better prognosis and survival rate. However, higher production of XPO4 in paracancerous liver tissue suggested a worse prognosis. All the results above provide new insights into better understanding biological indicators, such as XPO4, TGFβ1, ANGPTL4 and elF5A2, in the prediction and evaluation of liver cancer, as well as signaling pathways in the control of liver cancer. XPO4 and TGFβ1 may serve as useful markers to evaluate the size and prognosis of liver cancer.
Project description:Hepatocellular carcinoma (HCC) is a common malignant tumor in the clinic. Although there are increasing numbers of available treatment methods, their therapeutic effects are not satisfactory. The clinical indicators commonly used to predict the prognosis of HCC include tumor size, degree of cirrhosis, degree of tumor differentiation and tumor microvascular invasion; however, there are currently no molecular indicators that can predict the prognosis of HCC. Due to the differences in the progression of liver cancer among individuals, there is a growing need for prognostic biomarkers to accurately stratify patients for appropriate risk-adaptive treatment. The DNA topoisomerase 2-? (TOP2A) gene, which is located on human chromosome 17, encodes DNA topoisomerase II?. Previous studies have demonstrated that TOP2A indicates a poor prognosis in patients with various types of tumors, but no such studies are currently available on HCC. By analyzing the differential expression of TOP2A in 50 pairs of tumor and paracancerous tissue samples in The Cancer Genome Atlas (TCGA) database, the present study revealed that the expression of TOP2A was significantly higher in tumor tissue compared with that in paracancerous tissue (P=6.319×10-16). In the collected clinical samples, the mRNA expression levels of TOP2A were significantly upregulated in HCC tumor tissues compared with those in the paracancerous tissues (P=6.40×10-3), suggesting that TOP2A was associated with the occurrence and development of liver cancer. In addition, the associations between TOP2A expression, clinicopathological features and prognosis were analyzed using a multi-center large sample dataset from TCGA database, and the results demonstrated that high expression of TOP2A was associated with a higher T stage, poorer clinical stage and higher histological grade compared with those in patients with low TOP2A expression. High expression of TOP2A was also identified to be associated with a poor prognosis of HCC, particularly in Asian populations. These results suggested that high expression of TOP2A in HCC tissues may be closely associated with tumor progression and metastasis, which may be used as a biological indicator to predict tumor prognosis in clinical practice.
Project description:To support our research of colon cancer in human genome, we conducted massively parallel pyrosequencing of mRNAs (RNA-seq) using normal,paracancerouse and cancerous human colon tissues. We obtained a total of 29.9M reads from normal,33.0M reads from paracancerous and 36.5M reads from cancerous.The RNA-seq data derived from the sample illustrated the differencially expreesion genes among normal,paracancerous and cancerous colon tissues of human. 3 samples examined: normal tissue, paracancerous tissue, cancerous tissue.
Project description:In order to support our research of bladder cancer in human genome, we conducted massively parallel pyrosequencing of mRNAs (RNA-Seq) using normal, paracancerouse and cancerous human bladder tissues. We obtained a total of 30.0 million read pairs from normal, 33.1 million read pairs from paracancerous and 36.5 million read pairs from cancerous. The RNA-Seq data derived from the sample illustrated the differencially expression genes among normal, paracancerous and cancerous bladder tissues of human. 3 samples examined: normal tissue, paracancerous tissue, cancerous tissue.
Project description:Hepatocellular carcinoma (HCC) is one of the most diagnosed malignancies and a leading cause of cancer-related mortality globally. This is exacerbated by its highly aggressive phenotype, and limitation in early diagnosis and effective therapies. The SUMO-activating enzyme subunit 1 (SAE1) is a component of a heterodimeric small ubiquitin-related modifier that plays a vital role in SUMOylation, a post-translational modification involving in cellular events such as regulation of transcription, cell cycle and apoptosis. Reported overexpression of <i>SAE1</i> in glioma in a stage-dependent manner suggests it has a probable role in cancer initiation and progression. In this study, hypothesizing that <i>SAE1</i> is implicated in HCC metastatic phenotype and poor prognosis, we analyzed the expression of <i>SAE1</i> in several cancer databases and to unravel the underlying molecular mechanism of SAE1-associated hepatocarcinogenesis. Here, we demonstrated that <i>SAE1</i> is over-expressed in HCC samples compared to normal liver tissue, and this observed <i>SAE1</i> overexpression is stage and grade-dependent and associated with poor survival. The receiver operating characteristic analysis of <i>SAE1</i> in TCGA-LIHC patients (<i>n</i> = 421) showed an AUC of 0.925, indicating an excellent diagnostic value of <i>SAE1</i> in HCC. Our protein-protein interaction analysis for SAE1 showed that SAE1 interacted with and activated oncogenes such as <i>PLK1</i>, <i>CCNB1</i>, <i>CDK4</i> and <i>CDK1</i>, while simultaneously inhibiting tumor suppressors including <i>PDK4</i>, <i>KLF9</i>, <i>FOXO1</i> and <i>ALDH2</i>. Immunohistochemical staining and clinicopathological correlate analysis of SAE1 in our TMU-SHH HCC cohort (<i>n</i> = 54) further validated the overexpression of SAE1 in cancerous liver tissues compared with 'normal' paracancerous tissue, and high SAE1 expression was strongly correlated with metastasis and disease progression. The oncogenic effect of upregulated <i>SAE1</i> is associated with dysregulated cancer metabolic signaling. In conclusion, the present study demonstrates that SAE1 is a targetable cancer metabolic biomarker with high potential diagnostic and prognostic implications for patients with HCC.
Project description:PURPOSE: We investigated the impact of promoter methylation on APC protein expression in patients with hepatocellular carcinoma (HCC). MATERIALS AND METHODS: 50 patients [HCC (n=19), liver metastasis (n=19), cholangiocellular cancer (n=7), and benign liver tumors (n=5)] were studied for methylation using Methylight analysis. APC mutation was investigated by protein truncation test and direct sequencing of genomic DNA. The protein expression was evaluated by immunohistochemistry and Western blot analysis. RESULTS: The APC promoter was hypermethylated in 81.8% of non-cancerous liver tissue samples. All HCC samples and ten patients with liver metastasis (52.6%) exhibited APC promoter methylation. The degree of methylation was significantly higher in samples from HCC compared to the non-cancerous liver tissue samples (63.1% vs. 24.98%; p=0.001). The level of APC protein expression was significantly reduced in HCC samples compared to that of the corresponding non-tumor liver tissue (p<0.05). CONCLUSIONS: Promoter methylation of the APC gene seems to be of significance in hepatocarcinogenesis and results in reduced protein expression in HCC. Interestingly, APC promoter methylation is also present in the vast majority of non-cancerous liver tissue whose (patho)physiological function remains unresolved.
Project description:AIM:To study the expression profiles of HBsAg, HBcAg, p21WAF1/CIP1 (p21), Rb genes in hepatocellular carcinoma (HCC) and to investigate their roles in the hepatocar-cinogenesis. METHODS:HCC tissue microarray containing 120-min tissues of 40 HCC cases was constructed. HBsAg, HBcAg, p21 and Rb proteins were immunohistochemically stained by streptavidin-peroxidase conjugated method (S-P). The expression loss of these genes in cancerous, para-cancerous tissues and adjacent normal liver tissues of 40 HCCs were comparatively examined. RESULTS:The positive rate of HBsAg expression in cancerous tissues of 40 HCCs was 7.5%, which was lower than that in para-cancerous and adjacent normal liver tissues (chi2 =12.774, P < 0.01; chi2 = 18.442, P < 0.01). The positive rate of HBcAg expression in cancerous tissues of 40 HCCs was 20.0%, which was also lower than that in para-cancerous and adjacent normal liver tissues (chi2 = 9.482, P < 0.01; chi2 = 14.645, P < 0.01). p21 protein deletion rate in cancerous tissues of 40 HCCs was 27.5%, which was higher than that in para-cancerous and adjacent normal liver tissues (chi2 = 7.439, P < 0.01; chi2 = 11.174, P < 0.01). p21 protein deletion correlated remarkably with the pathological grade of HCC (chi2 = 0.072, P < 0.05). Rb protein deletion rate in cancerous tissues of 40 HCCs was 42.5%, which was also higher than that in para-cancerous and adjacent normal liver tissues (chi2 = 10.551, P < 0.01; chi2 = 18.353, P < 0.01). Rb protein deletion rate did not correlate remarkably with tumor size or pathological grade of HCC (chi2 = 0.014, P > 0.05; chi2 = 0.017, P > 0.05). CONCLUSION:Expression deletion of HBsAg, HBcAg, p21 and Rb proteins in HCCs may play important roles in the carcinogenesis of HCC. Tissue microarray is an effective high-throughput technique platform for cancer research.
Project description:Recent studies have shown that metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) was overexpressed in many human solid cancers, however, its roles in plasma of hepatocellular carcinoma (HCC) patients were unclear. The aim of this study was to investigate the significance of plasma MALAT1 levels in HCC patients. Plasma samples were collected from pre-operative HCC, hepatic disease patients, and healthy controls, and tissue samples from HCC patients and colorectal cancer patients with liver metastasis. Plasma and tissue MALAT1 levels were measured. Plasma MALAT1 levels were progressively and significantly higher in HCC patients than hepatic disease patients, and higher in hepatic disease patients than healthy controls. The expression of MALAT1 in HCC tissue was slightly higher than that in paired non-cancerous liver tissue, but not significant. The expression of MALAT1 in the non-cancerous liver tissue of 20 HCC patients was significantly higher than that in normal liver tissue of 13 colorectal cancer patients. In contrast, plasma MALAT1 levels were significantly low in HCC patients with hepatitis B infection, and significantly high in patients with liver damage B or liver cirrhosis. In a receiver-operator curve analysis of HCC and hepatic disease patients, the cut-off value of plasma MALAT1 was 1.60 and the area under the curve was 0.66. Plasma MALAT1 levels were not correlated with α-fetoprotein or protein induced by vitamin K absence II, whereas sensitivity and specificity for the detection of HCC with the combination of MALAT1, α-fetoprotein, and protein induced by vitamin K absence II were 88.6% and 75%, respectively. In conclusion, the plasma MALAT1 level is associated with liver damage, and has clinical utility for predicting development of HCC.
Project description:As the leading malignancy among women, breast cancer is a serious threat to the life and health of women. In this context, it is of particular importance that a proper therapeutic target be identified for breast cancer treatment. We collected the pathological tissues of 80 patients, with the view to discovering appropriate molecular targets for the treatment of breast cancer, this paper analyzes the expressions of ZNF436, ?-catenin, EGFR and CMTM5 in breast cancer tissues, as well as their correlations with breast cancer in combination with the clinicopathologic characteristics of studied patients. Immunohistochemistry (IHC) was utilized to detect the expression levels of ZNF436, ?-catenin, EGFR and CMTM5 in cancerous and paracancerous tissues of breast cancer patients. The expression levels of ZNF436, ?-Catenin and EGFR in breast cancer tissues were significantly greater than those in paracancerous tissues in this study (p<0.05), while CMTM5 was highly expressed in paracancerous tissues (p<0.05). Additionally, the correlation of the expressions of such indicators with the staging, differentiation and lymphatic metastasis of breast cancer, were also found to be statistically significant at the level p<0.05. The different expression levels of ZNF436, ?-catenin, EGFR and CMTM5 in breast cancer and paracancerous tissues open up the possibility of utilizing them as molecular markers for breast cancer. These findings provide a theoretical basis for targeted molecular therapies for breast cancer, and hence carry a significant practical significance.
Project description:Germinal center kinase-like kinase (GLK) is a key controller of autoimmunity. In this study, we assessed the clinical relevance and tumorigenic effects of GLK in hepatocellular carcinoma (HCC). Using immunohistochemistry, we showed that the GLK proportion score increased in both cancerous and adjacent non-cancerous liver tissue from patients with HCC recurrence. A Kaplan-Meier analysis revealed that patients with a wide distribution of GLK in non-cancerous liver tissue had a higher rate of HCC recurrence than those with very low or no GLK expression. Multivariate Cox regression analyses indicated that a high GLK proportion score in non-cancerous liver tissue was an independent predictor of early HCC recurrence after resection. Lentiviral vector-mediated overexpression of GLK activated the nuclear factor kappa B (NFκB) signaling cascade and accelerated cell cycle progression in primary human hepatocytes, thereby promoting proliferation. An increase in GLK expression coincided with NFκB activation and enhanced expression of proliferating cell nuclear antigen in HCC tissue. Our findings demonstrate a potential hepatocarcinogenic effect of GLK and the feasibility of using GLK to predict early HCC recurrence.
Project description:BACKGROUND:Augmenter of liver regeneration (ALR) is an important polypeptide that participates in the process of liver regeneration. Two forms of ALR proteins are expressed in hepatocytes. Previous data have shown that ALR is essential for cell survival and has potential antimetastatic properties in hepatocellular carcinoma (HCC). AIMS:The study aimed to evaluate the expression levels of two forms of ALR proteins in HCC and their possible significance in HCC development. METHODS:Balb/c mouse monoclonal antibody against ALR protein was prepared in order to detect the ALR protein in HCC by Western blotting and immunohistochemistry. ALR mRNA expression levels were measured by real-time polymerase chain reaction in HCC tissues and compared to paracancerous liver tissues in 22 HCC patients. RESULTS:ALR mRNA expression in HCC liver tissues (1.51×10(6) copies/?L) was higher than in paracancerous tissues (1.04×10(4) copies/?L). ALR protein expression was also enhanced in HCC liver tissues. The enhanced ALR protein was shown to be 23 kDa by Western blotting. Immunohistochemical analysis showed that the 23 kDa ALR protein mainly existed in the hepatocyte cytosol. CONCLUSION:The 23 kDa ALR protein was highly expressed in HCC and may play an important role in hepatocarcinogenesis.