Establishment and characterization of cell lines constitutively expressing hepatitis B virus X-protein

Lee, Young Ik; Bong, Yong Sik; Poo, Ha Ryoung; Lee, Yoon Ik; Park, Jong Gu; Oh, Su One; Sohn, Mi Jin; lee, Sook; Park, Ui Sun; Kim, Nam Soon; Hyun, Sang Won

doi:10.1016/s0378-1119(97)00603-3

Cited by 17 publications

(8 citation statements)

References 18 publications

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“…Hepatoma cells and HBx-transfected hepatoma cells (20) were maintained in Dulbecco's modified Eagle's medium (Invitrogen) supplemented with 10% heat-inactivated fetal bovine serum (JBI, Daegu, Korea), 2 mM glutamine, 100 units/ml penicillin, and 100 mg/ml streptomycin. The in situ cell death detection kit for the TUNEL assay was from Roche Applied Science.…”

Section: Cell Lines and Materials-mentioning

confidence: 99%

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Human Hepatitis B Virus-X Protein Alters Mitochondrial Function and Physiology in Human Liver Cells

Lee

Hwang

et al. 2004

Journal of Biological Chemistry

160

137

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The hepatitis B virus-X protein (HBx) regulates fundamental aspects of mitochondrial physiology. We show that HBx down-regulates mitochondrial enzymes involved in electron transport in oxidative phosphorylation (complexes I, III, IV, and V) and sensitizes the mitochondrial membrane potential in a hepatoma cell line. HBx also increases the level of mitochondrial reactive oxygen species and lipid peroxide production. HBx does not activate apoptotic signaling, although it sensitizes hepatoma cells to apoptotic signaling, which is dependent on reactive oxygen species. Increased intrahepatic lipid peroxidation in HBx transgenic mice demonstrated that oxidative injury occurs as a direct result of HBx expression. Therefore, we conclude that mitochondrial dysfunction is a crucial pathophysiological factor in HBx-expressing hepatoma cells and provides an experimental rationale in the investigation of mitochondrial function in rapidly renewed tissues, as in hepatocellular carcinomas.

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Section: Cell Lines and Materials-mentioning

confidence: 99%

“…Phosphorylation-Human hepatoma cell lines expressing the HBx protein in HepG2 (1X, 3X, and 4X) and HepG3B (6X and 8X) cells were established and characterized (20). By using these cell lines we screened the genes that are expressed differentially by HBx.…”

Section: The Hbx Protein Down-regulates Mitochondrial Enzymes Involvementioning

confidence: 99%

Human Hepatitis B Virus-X Protein Alters Mitochondrial Function and Physiology in Human Liver Cells

Lee

Hwang

et al. 2004

Journal of Biological Chemistry

160

137

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“…Transfected HBV-X gene integration and expression in Hep3B-6X and 8X cell lines were detected by Southern and Northern blotting and its biological activity were detected by chloramphenicol acetyl transferase (CAT) assay using various kinds of promoter-CAT reporter systems (Lee et al, 1998a). HBV-X protein expression in these cell lines were also con®rmed by immunouorescence microscopy.…”

Section: Hbv-x Prolongs the G1/s Transition Of The Hepatoma Cellsmentioning

confidence: 99%

“…HBV-X protein expression in these cell lines were also con®rmed by immunouorescence microscopy. The HBV-X transfectant enhanced AP-1 transcription factor binding to its target DNA (Lee et al, 1998a). Therefore, Hep3B-6X and 8X cell lines are not only stable but also have speci®c biological functions.…”

Section: Hbv-x Prolongs the G1/s Transition Of The Hepatoma Cellsmentioning

confidence: 99%

Hepatitis B virus-X protein upregulates the expression of p21wafl/cipl and prolongs G1→S transition via a p53-independent pathway in human hepatoma cells

Park

Lee

et al. 2000

Oncogene

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Progression through the cell cycle is controlled by the induction of cyclins and activation of cognate cyclindependent kinases. The human hepatitis B virus-X (HBV-X) protein functions in gene expression alterations, in the sensitization of cells to apoptotic killing and deregulates cell growth arrest in certain cancer cell types. We have pursued the mechanism of growth arrest in Hep3B cells, a p53-mutant human hepatocellular carcinoma (HCC) cell line. In stable or transient HBV-X transformed Hep3B cells, HBV-X increased protein and mRNA levels of the cyclin-dependent kinase inhibitor (CDKI) p21 wa¯/cipl , increased binding of p21 wa¯/cipl with cyclin-dependent kinase 2 (CDK2), markedly inhibited cyclin E and CDK2 associated phosphorylation of histone H1 and induced the activation of a p21 promoter reporter construct. By using p21 promoter deletion constructs, the HBV-X responsive element was mapped to a region between 71185 and 71482, relative to the transcription start site. Promoter mutation analysis indicated that the HBV-X responsive site coincides with the ets factor binding sites. These data indicate that in human hepatocellular carcinoma cells, HBV-X can circumvent the loss of p53 functions and induces critical downstream regulatory events leading to transcriptional activation of p21 wa¯/cipl . As a consequence, there is an increased chance of acquisition of mutations which can enhance the genesis of hepatomas. Our results also emphasize the chemotherapeutic potential of p21 wa¯/cipl inhibitors, particularly in the HBV-X infected hepatoma which lacks functional p53. Oncogene (2000) 19, 3384 ± 3394.

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“…Many human HCC cell lines have been established for the studies on etiological factors of HCC, such as hepatitis B virus (HBV) [9][10][11][12][13][14] , hepatitis C virus (HCV) [15] , hemochromatosis [16] , thorotrast [17] , for the study of alpha fetoprotein (AFP) [18,19] , and for other studies [20][21][22][23][24][25][26][27][28][29][30] . Unfortunately, of the above human HCC cell lines, metastatic potential was rarely mentioned or demonstrated.For animal HCC cell lines, the establishment of such cell lines in rat [31] , in woodchuck [32][33][34] , and in chicken [35] have been reported.…”

Section: Hepatocellular Carcinoma Cell Linementioning

confidence: 99%

Metastatic human hepatocellular carcinoma models in nude mice and cell line with metastatic potential

Tang

Sun²,

Tian³

et al. 2001

WJG

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Metastatic human HCC model is needed for the studies on mechanism and intervention of metastatic recurrence. By using orthotopic implantation of histologically intact tissues of 30 surgical specimens, a patient-like metastatic model of human HCC in nude mice (LCI-D20) and a low metastatic model of human HCC in nude mice (LCI-D35) have been established. All mice with transplanted LCI-D20 tumors exhibited e x t r e m e l y h i g h m e t a s t a t i c a b i l i t y i n c l u d i n g spontaneous metastasis to liver, lungs, lymph nodes and peritoneal seeding. Remarkable difference was also found in expression of some of the invasiveness related genes and growth factors between the LCI-D20 and LCI-D35 tumors. PAI-1 increased gradually following tumor progression in LCI-D20 model, and correlated with tumor size and AFP level. Phasic expression of tissue intercellular adhesion molecule-1 in this model was also observed. Using corneal micropocket model, it was demonstrated that the vascular response induced by LCI-D20 tumor was stronger than that induced by LCI-D35 tumor. Similar report on metastatic human HCC model in nude mice and human HCC cell line with metastatic potential was rarely found in the literature. This LCI-D20 model has been widely used for the studies on intervention of metastasis, including anti-angiogenesis, antisense approach, metallopro teinase inhibitor, differentiation inducer, etc. It is concluded that the establishment of metastatic human HCC model in nude mice and human HCC cell line with metastatic potential will provide important models for the in vivo and in vitro study of HCC invasiveness, angiogenesis as well as intervention of HCC recurrence.Subject headings hepatocellular carcinoma; metastasis; metastatic model; nude mice; cell line; experimental intervention; angiogenesis

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Establishment and characterization of cell lines constitutively expressing hepatitis B virus X-protein

Cited by 17 publications

References 18 publications

Human Hepatitis B Virus-X Protein Alters Mitochondrial Function and Physiology in Human Liver Cells

Human Hepatitis B Virus-X Protein Alters Mitochondrial Function and Physiology in Human Liver Cells

Hepatitis B virus-X protein upregulates the expression of p21wafl/cipl and prolongs G1→S transition via a p53-independent pathway in human hepatoma cells

Metastatic human hepatocellular carcinoma models in nude mice and cell line with metastatic potential

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