• Genomics & Informatics
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• 제8권1호
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• pp.9-18
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• 2010

Integrins are transmembrane receptor proteins that mediate cell-cell adhesion and cell-extracellular matrix (ECM) adhesion. The deregulation of cell-ECM adhesion and the abnormal expression of beta1 (${\beta}1$) integrins (ITGB1s) are involved in tumor development and metastasis. In the liver, the expression of integrins and ECM proteins can be a cause of hepatocellular carcinoma (HCC) development. We performed direct DNA sequencing of 24 individuals, and identified 23 sequence variants of ITGB1 polymorphisms. Among these 23 variants, 7 common variants were selected based on frequencies and linkage disequilibrium, and then genotyped in a larger-scale group of subjects (n=1,103). The genetic associations of ITGB1 polymorphisms with the clearance of HBV and HCC outcome of HBV patients were analyzed using logistic regression models and Cox relative hazard models. Although there was no significant association observed between the polymorphisms and the HCC outcome of HBV patients, the second most common haplotype (ITGB1 haplotype-2 [C-C-C-C-T-C-T]) was putatively associated with HBV clearance (OR=0.75, p=0.008 and $P^{corr}=0.05$). The minor allele frequency (MAF) of ITGB1 haplotype -2 of the spontaneously recovered (SR) group was significantly higher than that of the chronic carrier group (CC) (freq. = 0.248 vs. 0.199). The information derived from this study could be valuable for understanding the genetic factors involved in the clearance of HBV.

• Journal of Yeungnam Medical Science
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• 제25권1호
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• pp.1-18
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• 2008

1980년대부터 시행된 HBV 감염의 예방 백신접종으로 HBV 감염율이 감소되고 1997년부터 시작된 핵산유사체인 LMV의 항바이러스치료 효과로 인하여 CHB 환자에서 HBV 증식 억압이 가능하게 되면서 10~20여년 동안 HCC 발생율이 현저히 감소된다는 보고가 증가하고 있으며 21세기 전반기에는 1980년대 부터인 백신접종가능 시기 이후의 출생자들이 성인이 되는 시점이 되면 HBV 감염율 감소로 동반된 HCC 발생율의 현저한 감소를 예측하고 있다. 이미 백신 개발 이 전에 또는 백신접종의 기회를 얻지 못하고 이미 영유아 시기에 감염되어 CHB에 이환된 환자들의 간경변증, HCC 등 만성 합병증을 치료하기 위해서는 적극적인 항바이러스치료가 필요하였으나 1980년대 후반에 유일하게 CHB 치료제로 인정된 인터페론은 치료 적응증이나 그 효과에 있었으나 극소수의 제한된 환자에서 제한된 효과만이 입증되었으며 특히 역학적 차이로 인한 바이러스학적, 개체 면역학적 조건에 따라 전혀 고려되지 않는 경우가 많고 수직감염자가 대다수인 우리나라를 포함한 대부분의 고감역지역에서는 우선 선택되지 않는 약제였으며 장기 효과에 대해서는 이십여년이 지난 시점에도 확실하지 않은 상황에서, 1997년부터 극적인 항바이러스 효과를 나타내는 LMV을 투약할 수 있게 되었다. LMV의 HCC 발생 억제효과를 높이기 위해 개선해야 할 점은 필수적인 장기투약에 의해 발생되는 내성 바이러스의 출현이며 이는 간기능뿐 만아니라 HCC 발생의 위험도 높일 수 있다는 보고도 있다. LMV 이후 처방 허가된 ADV, telbivudine(LdT), ETV, CLV, TDF 등도 장기 투약시 내성바이러스 발생이 가능하나 내성 발생을 줄이기 위해서는 적절한 약의 선택과 새로운 약제의 개발과 동시에 치료 원칙의 정립이 필요하다. 그러나 이러한 문제점이 있음에도 불구하고 항바이러스치료로 CHB의 진행 및 만성 합병증 발생의 빈도가 감소되고 있으며 HCC 발생 역시 감소될 것이며, 그 효과를 더 높이기 위해서는 내성변이종 발생 문제를 해결해야 하며 치료대상, 치료기준, 약제의 선택과 방법 및 면밀한 검사관리가 필요하다. 그러나 무엇보다 중요한 것은 일단 HBV에 감염되면 과거감염이건 현재 감염이건 현재의 핵산유사체 제제로 HBV DNA 증식을 억제하더라도 간조직내 cccDNA와 융합된 HBV DNA를 보유할 수 있으며 그 증거로는 HBsAg이 음성인 HCC 환자의 혈청이나 간조직에서 HBV DNA가 발견되는 빈도가 높다는 증거가 무수히 많고, 상황에 따라서는 간질환의 원인이 된다는 보고가 다수 있으므로 가장 완벽하고 확실한 예방법인 HBV 백신 접종이 무엇보다 필수적이며 전세계 대부분의 HCC 발생을 예방할 수 있다. 또한 현재 제시되고 있는 항바이러스제 치료기준은 혈청 간세포괴사치 상승을 조건으로 제한하는데 혈청 ALT치나 HBeAg 상태에 무관하게 간 조직의 상태나 HBV DNA치의 증가와 비례해서 간경변이나 HCC 발생의 위험이 높아진다는 사실이 중요하다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권1호
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• pp.225-229
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• 2012

Background: Males have a higher prevalence of hepatocellular carcinoma (HCC) than females in general, but the reasons for the sex disparity are still obscure. DNA copy number alteration (CNA) is a major feature of solid tumors including HCC, but whether CNA plays a role in sex-related differences in HCC development has never been evaluated. Methods: High-resolution array comparative genomic hybridization (CGH) was used to examine 17 female and 46 male HCC patients with chronic hepatitis B virus (HBV) infection in Shanghai, China. Two-tailed Fisher's exact or ${\chi}^2$ tests was used to compare CNAs between females and males. Results: The overall frequencies and patterns of CNAs in female and male cases were similar. However, female HCC tumors presented more copy number gains compared to those in males on 1q21.3-q22 (76.5% vs. 37.0%, P = 0.009), 11q11 (35.3% vs. 0.0%, P = 0.0002) and 19q13.31-q13.32 (23.5% vs. 0.0%, P = 0.004), and loss on 16p11.2 (35.3% vs. 6.5%, P = 0.009). Relative to females, male cases had greater copy number loss on 11q11 (63.0% vs. 17.6%, P = 0.002). Further analyses showed that 11q11 gain correlated with 19q13.31-q13.32 gain (P = 0.042), 11q11 loss (P = 0.011) and 16p11.2 loss (P = 0.033), while 1q21.3-q22 gain correlated with 19q13.31-q13.32 gain (P = 0.046). Conclusions: These findings suggest that CNAs may play a role in sex-related differences in HBVassociated HCC development.

• Asian Pacific Journal of Cancer Prevention
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• 제15권20호
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• pp.8623-8629
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• 2014

Background: As an important component of the NDC80 kinetochore complex, NUF2 is essential for kinetochore-microtubule attachment and chromosome segregation. Previous studies also suggested its involvement in development of various kinds of human cancers, however, its expression and functions in human hepatocellular carcinoma (HCC) are still unclear. Materials and Methods: In the present study, we aimed to test the hypothesis that NUF2 is aberrant in human HCCs and associated with cell growth. Results: Our results showed significantly elevated expression of NUF2 in human HCC tissues compared to adjacent normal tissues, and high expression of NUF2 in HCC cell lines. Using lentivirus-mediated silencing of NUF2 in HepG2 human HCC cells, we found that NUF2 depletion markedly suppressed proliferation and colony formation capacity in vitro, and dramatically hampered tumor growth of xenografts in vivo. Moreover, NUF2 silencing could induce cell cycle arrest and trigger cell apoptosis. Additionally, altered levels of cell cycle and apoptosis related proteins including cyclin B1, Cdc25A, Cdc2, Bad and Bax were also observed. Conclusions: In conclusion, these results demonstrate that NUF2 plays a critical role in the regulation of HCC cell proliferation and apoptosis, indicating that NUF2 may serve as a potential molecular target for therapeutic approaches.

• Asian Pacific Journal of Cancer Prevention
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• 제17권7호
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• pp.3369-3375
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• 2016

MicroRNAs, a novel class of small non-coding RNAs, are key players in many cellular processes, including cell proliferation, differentiation, invasion and regeneration. Tissue and circulatory microRNAs could serve as useful clinical biomarkers and deregulated expression levels have been observed in various cancers. Gene variants may alter microRNA processing and maturation. Thus, we aimed to investigate the association of MIR-196a2 rs11614913 (C/T), MIR-499a rs3746444 (A/G) polymorphisms and their combination with cancer susceptibility in an Egyptian population. Sixty five renal cell carcinoma (RCC) and 60 hepatocellular carcinoma (HCC) patients and 150 controls were enrolled in the study. They were genotyped using real-time polymerase chain reaction technology. Both $miR-196a2^*T$ and $miR-499a*G$ were associated with RCC risk, but only $miR-196a^*T$ was associated with HCC development. Carriage of the homozygote combinations ($MIR196a2^*TT+MIR499a^*AA$) and ($MIR196a2^*CC+MIR499a^*GG$) was associated with 25 and 48 fold elevation of likelhood to develop RCC, respectively. The miR-196a2 SNP was also linked with larger tumor size in RCC and advanced tumor stage in HCC. miR-196a2 and miR-499a combined genotypes were associated with RCC and HCC. Further functional analysis of SNPs is required to confirm relationships between genotypes and phenotypes.

• BMB Reports
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• 제50권1호
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• pp.1-2
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• 2017

Acquiring a selective growth advantage by breaking the proliferation barrier established by gatekeeper genes is a centrally important event in tumor formation. Removal of the mammalian Hippo kinase Mst1 and Mst2 in hepatocytes leads to rapid hepatocellular carcinoma (HCC) formation, indicating that the Hippo signaling pathway is a critical gatekeeper that restrains abnormal growth in hepatocytes. By rigorous genetic approaches, we identified an interacting network of the Hippo, Wnt/${\beta}$-catenin and Notch signaling pathways that control organ size and HCC development. We found that in hepatocytes, the loss of Mst1/2 leads to the activation of Notch signaling, which forms a positive feedback loop with Yap/Taz (transcription factors controlled by Mst1/2). This positive feedback loop results in severe liver enlargement and rapid HCC formation. Blocking the Yap/Taz-Notch positive feedback loop by Notch inhibition in vivo significantly reduced the Yap/Taz activities, hepatocyte proliferation and tumor formation. Furthermore, we uncovered a surprising inhibitory role of Wnt/${\beta}$-catenin signaling to Yap/Taz activities, which are important in tumor initiation. Genetic removal of ${\beta}$-catenin in the liver of the Mst1/2 mutants significantly accelerates tumoriogenesis. Therefore, Wnt/${\beta}$-catenin signaling, known for its oncogenic property, exerts an unexpected function in restricting Yap/Taz and Notch activities in HCC initiation. The molecular interplay between the three signaling pathways identified in our study provides new insights in developing novel therapeutic strategies to treat liver tumors.

• Journal of Microbiology and Biotechnology
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• 제29권8호
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• pp.1281-1287
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• 2019

Nattokinase (NK, E.C. 3.4.21.62) is a serine protease produced by Bacillus subtilis natto that shows promise for the treatment of thrombotic disease. In this study, we assessed the effects of NK on the development of hepatocellular carcinoma (HCC), a principal malignancy of the liver that causes morbidity and mortality worldwide. Crude extracts of NK (NCE) were isolated from fermentation medium by centrifugation and separated into three fractions (<10 K, 100~30 K and >30K). Orthotopic HCC mouse models were established and NCE was administered by oral gavage. H&E staining was performed to examine the pathology of HCC livers. Immunohistochemistry and immunofluorescence were used to evaluate FOXM1, CD31, CD44 and vimentin expression in the liver. Compared to PBS groups, NCE increased the survival rates of HCC-bearing mice to 31% and decreased ascites. Low-intensity ultrasound imaging showed that the hypoechoic mass area was lower in NCE-treated mice and that tumor growth significantly decreased. IHC staining showed that the expression of FOXM1 was inhibited by NCE treatment. Immunofluorescence results revealed lower levels of CD31, CD44 and vimentin in the NCE groups. Taken together, these data demonstrate that NCE from Bacillus subtilis natto improves survival and inhibits tumor growth in HCC mice.

• The Korean Journal of Physiology and Pharmacology
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• 제17권5호
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• pp.455-461
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• 2013

Retinoids regulate not only various cell functions including proliferation and differentiation but also glucose and lipid metabolism. After we observed a marked up-regulation of cellular retinol-binding protein-I (CRBP-I) in the liver of hepatitis B virus x antigen (HBx)-transgenic (HBx Tg) mice which are prone to hepatocellular carcinoma (HCC) and fatty liver, we aimed to evaluate retinoid pathway, including genes for the retinoid physiology, CRBP-I protein expression, and retinoid levels, in the liver of HBx Tg mice. We also assessed the effect of chronic metformin treatment on HCC development in the mice. Many genes involved in hepatic retinoid physiology, including CRBP-I, were altered and the tissue levels of retinol and all-trans retinoic acid (ATRA) were elevated in the liver of HBx Tg mice compared to those of wild type (WT) control mice. CRBP-I protein expression in liver, but not in white adipose tissue, of HBx Tg mice was significantly elevated compared to WT control mice while CRBP-I protein expressions in the liver and WAT of high-fat fed obese and db/db mice were comparable to WT control mice. Chronic treatment of HBx Tg mice with metformin did not affect the incidence of HCC, but slightly increased hepatic CRBP-I level. In conclusion, hepatic CRBP-I level was markedly up-regulated in HCC-prone HBx Tg mice and neither hepatic CRBP-I nor the development of HCC was suppressed by metformin treatment.

• Asian Pacific Journal of Cancer Prevention
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• 제16권5호
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• pp.2043-2049
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• 2015

Background: Serum Golgi protein 73 (GP73) as a novel and potential marker for diagnosing hepatocellular carcinoma (HCC) have been found to be elevated in HCC patients and associated with clinical variables representing tumor growth and invasiveness. The aim of this study was to prepare a pair of monoclonal antibodys (mAbs) against GP73 and develop a newly designed double-antibody sandwich enzyme-linked immunosorbent assay (s-ELISA), which would be used in the detection of serum GP73 (sGP73) as well as in the diagnosis of HCC. Materials and Methods: Produced by prokaryotic expression, the purified recombinant GP73 (rGP73), produced by prokaryotic expression, was used to immunize the Balb/c mice. Two hybridoma cell lines against GP73 were obtained by fusing mouse Sp2/0 myeloma cells with spleen cells from the immunized mice. The titers of anti-GP73 mAb reached 1:243,000. Western blotting analysis and Immunohistochemistry staining revealed that anti-GP73 mAb could recognize GP73 protein. The double-antibody s-ELISA was successfully established and validated by 119 HCC and 103 normal serum samples. Results: showed that the detection limit of this method could reach 1.56 ng/ml, and sGP73 levels in HCC group (mean=190.6 ng/ml) were much higher than those of in healthy controls (mean=70.92 ng/ml). Conclusions: Results of our study not only showed that sGP73 levels of HCC patients were significantly higher than those of healthy controls, but also indicated that the laboratory homemade anti-GP73 mAbs could be the optimal tool used in evaluating sGP73 levels, which would provide a solid foundation for subsequent clinical applications.

• Molecular & Cellular Toxicology
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• 제4권2호
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• pp.93-99
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• 2008

Human cyclin D3 gene (CCND3) located on 6p21.1 is important for the regulation of the G1-S phase transition of the cell cycle by modulating the activity of the cyclin-dependent kinases Cdk4 and Cdk6. Because little is known about the effect of cyclin D3 in various human cancers, we evaluated the intricate relationship between expression of cyclin D3 and the process of HCC development using immunohis tochemistry and TUNEL assay on 43 paraffin embedded tissues. Cyclin D3 immunoreactivity was more frequently observed in the tumors with high histologic grade and the tumors with metastasis, and more frequently expressed in HCCs with cirrhotic background and gain of 6p21.1 when compared with those with non-neoplastic tissue. Apoptotic cells were more common in tumor with cirrhotic background, amplification of 6p21.1 and expression of cyclin D3 when compared with HCCs with lower level of cyclin D3 expression. Also, we observed that some of the cyclin D3 positive cell and apoptotic cell were co-localized. From these results, it is suggested that over-expression of cyclin D3 may contribute to more rapid cell turn-over in the background of HCC, and balance between proliferation and apoptosis is a role in the progression of HCC with cirrhotic background.