Elimination of Cancer Stem-Like “Side Population” Cells in Hepatoma Cell Lines by Chinese Herbal Mixture “Tien-Hsien Liquid”

There are increasing pieces of evidence suggesting that the recurrence of cancer may result from a small subpopulation of cancer stem cells, which are resistant to the conventional chemotherapy and radiotherapy. We investigated the effects of Chinese herbal mixtureTien-Hsien Liquid(THL) on the cancer stem-like side population (SP) cells isolated from human hepatoma cells. After sorting and subsequent culture, the SP cells from Huh7 hepatoma cells appear to have higher clonogenicity and mRNA expressions of stemness genes such asSMO, ABCG2, CD133,β-catenin,andOct-4than those of non-SP cells. At dose of 2 mg/mL, THL reduced the proportion of SP cells in HepG2, Hep3B, and Huh7 cells from 1.33% to 0.49%, 1.55% to 0.43%, and 1.69% to 0.27%, respectively. The viability and colony formation of Huh7 SP cells were effectively suppressed by THL dose-dependently, accompanied with the inhibition of stemness genes, e.g.,ABCG2, CD133,andSMO. The tumorigenicity of THL-treated Huh7 SP cells in NOD/SCID mice was also diminished. Moreover, combination with THL could synergize the effect of doxorubicin against Huh7 SP cells. Our data indicate that THL may act as a cancer stem cell targeting therapeutics and be regarded as complementary and integrative medicine in the treatment of hepatoma.

Download Full-text

Inhibitory Effects of Lycopene on the Adhesion, Invasion, and Migration of SK-Hep1 Human Hepatoma Cells

Experimental Biology and Medicine ◽

10.1177/153537020623100313 ◽

2006 ◽

Vol 231 (3) ◽

pp. 322-327 ◽

Cited By ~ 56

Author(s):

Eun-Sun Hwang ◽

Hyong Joo Lee

Keyword(s):

Cell Growth ◽

Hepatoma Cell ◽

Hepatoma Cells ◽

Hepatoma Cell Line ◽

Dependent Manner ◽

Human Hepatoma ◽

Human Hepatoma Cells ◽

Human Hepatoma Cell ◽

Invasion And Migration ◽

And Migration

Lycopene, which is the predominant carotenoid in tomatoes and tomato-based foods, may protect humans against various cancers. Effects of lycopene on the adhesion, invasion, migration, and growth of the SK-Hep1 human hepatoma cell line were investigated. Lycopene inhibited cell growth in dose-dependent manners, with growth inhibition rates of 5% and 40% at 0.1 μM and 50 μM lycopene, respectively, after 24 hrs of incubation. Similarly, after 48 hrs of incubation, lycopene at 5 μM and 10 μM decreased the cell numbers by 30% and 40%, respectively. Lycopene decreased the gelatinolytic activities of both matrix metalloproteinase (MMP)-2 and MMP-9, which were secreted from the SK-Hep1 cells. Incubation of SK-Hep1 cells with 110 μM of lycopene for 60 mins significantly inhibited cell adhesion to the Matrigel-coated substrate in a concentration-dependent manner. To study invasion, SK-Hep1 cells were grown either on Matrigel-coated Transwell membranes or in 24-well plates. The cells were treated sequentially for 24 hrs with lycopene before the start of the invasion assays. Cell growth and death were assessed under the same conditions. The invasion of SK-Hep1 cells treated with lycopene was significantly reduced to 28.3% and 61.9% of the control levels at 5 μM and 10 μM lycopene, respectively (P < 0.05). In the migration assay, lycopene-treated cells showed lower levels of migration than untreated cells. These results demonstrate the antimetastatic properties of lycopene in inhibiting the adhesion, invasion, and migration of SK-Hep1 human hepatoma cells.

Download Full-text

Inhibiting Effects of Down-regulating of Fascin 1 on Proliferation and Migration in Hepatoma Cells

10.21203/rs.3.rs-942724/v1 ◽

2021 ◽

Author(s):

Wanglu Gu ◽

Guilan Wang ◽

Xinyang Zhang ◽

Li Chen ◽

Jiaming Zhou

Keyword(s):

Protein Expression ◽

Western Blotting ◽

Hepatoma Cell ◽

Hepatoma Cells ◽

Human Hepatoma Cell ◽

Specific Sequence ◽

Knock Down ◽

Huh7 Cells ◽

And Migration ◽

Proliferation And Migration

Abstract Objective: To investigate the inhibiting effects of fascin 1 gene knock-down on the proliferation and migration of hepatoma cells by means of small interfering RNA (siRNA).Methods: SiRNA targeting fascin 1 gene (si-fascin) and non-specific sequence siRNA (si-NC）were constructed and transfected into human hepatoma cell lines (HepG2 and Huh7) to down-regulate the expression of fascin 1. RT-qPCR, Western blotting, and Immunofluorescence technique were used to evaluate the efficiency of si-fascin. The proliferation and migration of cells were detected by MTT method and Transwell experiments, and the protein expression of genes related to proliferation and migration in cells were detected by Western blotting. The apoptosis and pseudopodia formation of cells were observed under scanning electron microscope (SEM).Results: Compared with human normal liver cells (LO2), the expressions of fascin 1 mRNA and protein were significantly higher in HepG2 and Huh7 cells. The expression of fascin 1 was overall inhibited in HepG2 and Huh7 cells transfected by the constructed four si-fascins, among which, fascin_siR3 had the highest inhibitory efficiency, therefore was selected in this study. In HepG2 and Huh7 cells transfected by si-fascin significant knock-down target gene expression, while reducing cell proliferation, migration and the formation of pseudopods, and causes reduced protein expression associated with proliferation and migration. Conclusion: This study further confirmed that fascin 1 gene has the function of promoting hepatoma cells proliferation and migration, suggesting that downregulating the expression of fascin 1 in hepatoma cells may be one of the strategies to intervene in liver cancer.

Download Full-text

PNPLA2 influences secretion of triglyceride-rich lipoproteins by human hepatoma cells

The Journal of Lipid Research ◽

10.1194/jlr.m090928 ◽

2019 ◽

Vol 60 (6) ◽

pp. 1069-1077 ◽

Cited By ~ 4

Author(s):

Apostolos Taxiarchis ◽

Hovsep Mahdessian ◽

Angela Silveira ◽

Rachel M. Fisher ◽

Ferdinand M. van’t Hooft

Keyword(s):

Lipid Droplet ◽

Physiological Role ◽

Hepatoma Cells ◽

Human Hepatoma ◽

Human Hepatoma Cells ◽

Triglyceride Metabolism ◽

Triglyceride Hydrolysis ◽

Triglyceride Accumulation ◽

Huh7 Cells ◽

Microscopy Analysis

Patatin-like phospholipase domain-containing proteins (PNPLAs) are involved in triglyceride hydrolysis and lipid-droplet homeostasis in mice, but the physiological significance of the PNPLAs for triglyceride metabolism in human hepatocytes is unclear. Here, we investigate the roles of PNPLA2, PNPLA3, and PNPLA4 in triglyceride metabolism of human Huh7 and HepG2 hepatoma cells using gene-specific inhibition methods. siRNA inhibition of PNPLA3 or PNPLA4 is not associated with changes in triglyceride hydrolysis, secretion of triglyceride-rich lipoproteins (TRLs), or triglyceride accumulation. However, PNPLA2 siRNA inhibition, both in the absence and presence of oleate-containing medium, or treatment with the PNPLA2 inhibitor Atglistatin reduced intracellular triglyceride hydrolysis and decreased TRL secretion. In contrast, PNPLA2 inhibition showed no effects on lipid-droplet homeostasis, which is the primary physiological function of PNPLA2 in nonhepatic tissues. Moreover, confocal microscopy analysis found no clear evidence for the localization of PNPLA2 around lipid droplets. However, significant colocalization of PNPLA2 with the endoplasmic reticulum marker protein disulfide-isomerase was found in HepG2 and Huh7 cells with Rcoloc values of 0.61 ± 0.06 and 0.81 ± 0.05, respectively. In conclusion, PNPLA2 influences TRL secretion, but is not involved in lipid-droplet homeostasis in human hepatoma cells, a physiological role that is quite distinct from the metabolic function of PNPLA2 in nonhepatic tissues.

Download Full-text

Targeting Autophagy Augments Berberine-Mediated Cell Death in Human Hepatoma Cells Harboring Hepatitis C Virus RNA

Cells ◽

10.3390/cells9040908 ◽

2020 ◽

Vol 9 (4) ◽

pp. 908 ◽

Cited By ~ 1

Author(s):

Chen-Jei Tai ◽

Alagie Jassey ◽

Ching-Hsuan Liu ◽

Cheng-Jeng Tai ◽

Christopher D. Richardson ◽

...

Keyword(s):

Cell Death ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Hepatoma Cell ◽

Treatment Strategies ◽

Hepatoma Cells ◽

Hcv Rna ◽

Human Hepatoma Cells ◽

Subgenomic Replicon ◽

Pharmacological Inhibition

Hepatocellular carcinoma (HCC), including hepatitis C virus (HCV)-induced HCC, is a deadly disease highly refractory to chemotherapy, thus requiring the continuous identification of novel treatment strategies. Berberine (BBR) has been previously reported to inhibit hepatoma cell growth, but the main type of cell death elicited by BBR, and whether the alkaloid can inhibit hepatoma cells carrying HCV genomes, is unclear. Herein, we show that BBR treatment induced a biphasic cell death irrespective of the presence of HCV subgenomic replicon RNA, first triggering apoptosis that then progressed to necrosis between 24 and 48 h post-treatment. Furthermore, BBR treatment potentiated the HCV replicon-induced reactive oxygen species (ROS) production, inhibition of which with an antioxidant attenuated the cell death that was elicited by BBR in these cells. Moreover, BBR dampened the autophagic response in HCV RNA-positive or negative hepatoma cells, and pharmacological inhibition of autophagy conversely augmented the BBR-induced cell death. Finally, BBR inhibited the growth of Huh-7 cells that were persistently infected with the full-length genome HCV particles, and concomitant pharmacological inhibition of autophagy potentiated the killing of these cells by BBR. Our findings suggest that combining BBR with the inhibition of autophagy could be an attractive treatment strategy against HCC, irrespective of the presence of the HCV genome.

Download Full-text

Liver-enriched transcription factors uncoupled from expression of hepatic functions in hepatoma cell lines.

Molecular and Cellular Biology ◽

10.1128/mcb.17.11.6311 ◽

1997 ◽

Vol 17 (11) ◽

pp. 6311-6320 ◽

Cited By ~ 14

Author(s):

D Chaya ◽

C Fougère-Deschatrette ◽

M C Weiss

Keyword(s):

Transcription Factors ◽

Hepatoma Cell ◽

Hepatoma Cells ◽

Nuclear Factor ◽

Hepatocyte Nuclear Factor ◽

Hepatic Differentiation ◽

Hepatoma Cell Lines ◽

Differentiated Cells ◽

Heritable Phenotype ◽

Nuclear Factor 1

Among the liver-enriched transcription factors identified to date, only expression of hepatocyte nuclear factor 4 (HNF4) and hepatocyte nuclear factor 1 (HNF1) is in strict correlation with hepatic differentiation in cultured rat hepatoma cells. Indeed, differentiated hepatoma cells that stably express an extensive set of adult hepatic functions express liver-enriched transcription factors, while dedifferentiated cells that have lost expression of all these hepatic functions no longer express HNF4 and HNF1. We describe a new heritable phenotype, designated as uncoupled, in which there is a spontaneous dissociation between the expression of these transcription factors and that of the hepatic functions. Cells presenting this phenotype, isolated from differentiated hepatoma cells, cease to accumulate all transcripts coding for hepatic functions but nevertheless maintain expression of HNF4 and HNF1. Transitory transfection experiments indicate that these two factors present in these cells have transcriptional activity similar to that of differentiated hepatoma cells. Characterization of the appropriate intertypic cell hybrids demonstrates that this new phenotype is recessive to the dedifferentiated state and fails to be complemented by differentiated cells. These results indicate the existence of mechanisms that inhibit transcription of genes coding for hepatocyte functions in spite of the presence of functional HNF4 and HNF1. Cells of the uncoupled phenotype present certain properties of oval cells described for pathological states of the liver.

Download Full-text

Pharmacological or TRIB3-Mediated Suppression of ATF4 Transcriptional Activity Promotes Hepatoma Cell Resistance to Proteasome Inhibitor Bortezomib

Cancers ◽

10.3390/cancers13102341 ◽

2021 ◽

Vol 13 (10) ◽

pp. 2341

Author(s):

Tiit Örd ◽

Daima Örd ◽

Minna U. Kaikkonen ◽

Tõnis Örd

Keyword(s):

Cell Death ◽

Er Stress ◽

Proteasome Inhibitor ◽

Transcriptional Activity ◽

Molecular Mechanisms ◽

Target Genes ◽

Hepatoma Cell ◽

Hepatoma Cells ◽

Cell Resistance ◽

Human Hepatoma Cells

The proteasome is an appealing target for anticancer therapy and the proteasome inhibitor bortezomib has been approved for the treatment of several types of malignancies. However, the molecular mechanisms underlying cancer cell resistance to bortezomib remain poorly understood. In the current article, we investigate how modulation of the eIF2α–ATF4 stress pathway affects hepatoma cell response to bortezomib. Transcriptome profiling revealed that many ATF4 transcriptional target genes are among the most upregulated genes in bortezomib-treated HepG2 human hepatoma cells. While pharmacological enhancement of the eIF2α–ATF4 pathway activity results in the elevation of the activities of all branches of the unfolded protein response (UPR) and sensitizes cells to bortezomib toxicity, the suppression of ATF4 induction delays bortezomib-induced cell death. The pseudokinase TRIB3, an inhibitor of ATF4, is expressed at a high basal level in hepatoma cells and is strongly upregulated in response to bortezomib. To map genome-wide chromatin binding loci of TRIB3 protein, we fused a Flag tag to endogenous TRIB3 in HepG2 cells and performed ChIP-Seq. The results demonstrate that TRIB3 predominantly colocalizes with ATF4 on chromatin and binds to genomic regions containing the C/EBP–ATF motif. Bortezomib treatment leads to a robust enrichment of TRIB3 binding near genes induced by bortezomib and involved in the ER stress response and cell death. Disruption of TRIB3 increases C/EBP–ATF-driven transcription, augments ER stress and cell death upon exposure to bortezomib, while TRIB3 overexpression enhances cell survival. Thus, TRIB3, colocalizing with ATF4 and limiting its transcriptional activity, functions as a factor increasing resistance to bortezomib, while pharmacological over-activation of eIF2α–ATF4 can overcome the endogenous restraint mechanisms and sensitize cells to bortezomib.

Download Full-text

Integrative Network Analysis Revealed Genetic Impact of Pyruvate Kinase L/R on Hepatocyte Proliferation and Graft Survival after Liver Transplantation

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/7182914 ◽

2021 ◽

Vol 2021 ◽

pp. 1-31

Author(s):

Zhengtao Liu ◽

Junsheng Zhao ◽

Wenchao Wang ◽

Hai Zhu ◽

Junjie Qian ◽

...

Keyword(s):

Liver Transplantation ◽

Pyruvate Kinase ◽

De Novo ◽

Hepatoma Cell ◽

Hepatoma Cells ◽

Arachidonic Acid Metabolism ◽

De Novo Lipogenesis ◽

Hepatocyte Proliferation ◽

Human Hepatoma Cells ◽

Genetic Impact

Background. Pyruvate kinase L/R (PKLR) has been suggested to affect the proliferation of hepatocytes via regulation of the cell cycle and lipid metabolism. However, its impact on the global metabolome and its clinical implications remain unclear. Aims. We aimed to clarify the genetic impact of PKLR on the metabolomic profiles of hepatoma cells and its potential effects on grafts for liver transplantation (LT). Methods. Nontargeted and targeted metabolomic assays were performed in human hepatoma cells transfected with lentiviral vectors causing PKLR overexpression and silencing, respectively. We then constructed a molecular network based on integrative analysis of transcriptomic and metabolomic data. We also assessed the biological functions of PKLR in the global metabolome in LT grafts in patients via a weighted correlation network model. Results. Multiomic analysis revealed that PKLR perturbations significantly affected the pyruvate, citrate, and glycerophospholipid metabolism pathways, as crucial steps in de novo lipogenesis (DNL). We also confirmed the importance of phosphatidylcholines (PC) and its derivative lyso-PC supply on improved survival of LT grafts in patients. Coexpression analysis revealed beneficial effects of PKLR overexpression on posttransplant prognosis by alleviating arachidonic acid metabolism of the grafts, independent of operational risk factors. Conclusion. This systems-level analysis indicated that PKLR affected hepatoma cell viability via impacts on the whole process of DNL, from glycolysis to final PC synthesis. PKLR also improved prognosis after LT, possibly via its impact on the increased genesis of beneficial glycerophospholipids.

Download Full-text

Immunofluorescent localization of type II insulin-like growth factor receptor in rat liver and hepatoma cells

Journal of Endocrinology ◽

10.1677/joe.0.1210221 ◽

1989 ◽

Vol 121 (2) ◽

pp. 221-NP ◽

Cited By ~ 23

Author(s):

M. A. Hartshorn ◽

C. D. Scott ◽

R. C. Baxter

Keyword(s):

Rat Liver ◽

Hepatoma Cell ◽

Rat Hepatocytes ◽

Hepatoma Cells ◽

Type Ii ◽

Adult Rat ◽

Hepatoma Cell Lines ◽

Igf Receptors ◽

Immunofluorescent Technique ◽

Igf Receptor

ABSTRACT We have used an immunofluorescent technique to localize type II insulin-like growth factor (IGF) receptors in rat liver, rat hepatocytes and three rat hepatoma cell lines (HTC, H-35 and 5123) using a polyclonal antibody (C-1) raised to purified rat liver type II IGF receptor. Specificity of the antiserum was confirmed by Western blotting of microsomal membranes prepared from hepatocytes and hepatoma cells which showed a single class of receptor in all cells, of Mr approximately 210 000 for hepatocytes, HTC and H-35 cells and approximately 220 000 for 5123 cells, on non-reduced, 4–15% polyacrylamide gradient gels. The specificity of the immunofluorescent technique was also verified by abolition of labelling after preincubation of antiserum with purified type II IGF receptor. Rat liver cryosections contained areas of juxtanuclear labelling in hepatocytes, consistent with the presence of type II IGF receptor in the Golgi region. Brightest immunofluorescence was seen in sections from fetal and neonatal rats with adult rat hepatocytes staining brightly only around central veins. Areas of labelling were also seen in connective tissue surrounding larger veins. Cultured adult rat hepatocytes and rat hepatoma cell lines also showed bright areas of juxtanuclear immunofluorescence, with HTC and H-35 cells staining more than 5123 and adult hepatocytes. Fetal rat hepatocytes in culture also labelled very brightly both in a juxtanuclear location and in small clusters over the cell, possibly on the cell surface. These observations indicate that type II IGF receptors are located predominantly on intracellular membranes and are most abundant in rapidly growing cells and tissues (such as fetal liver and hepatoma cells). Journal of Endocrinology (1989) 121, 221–227

Download Full-text

Resistance to paclitaxel in hepatoma cells is related to static JNK activation and prohibition into entry of mitosis

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00449.2011 ◽

2012 ◽

Vol 302 (9) ◽

pp. G1016-G1024 ◽

Cited By ~ 13

Author(s):

Sunyoung Chae ◽

Young Bae Kim ◽

Jong-Soo Lee ◽

Hyeseong Cho

Keyword(s):

Cell Death ◽

Anticancer Agents ◽

Kinase Activity ◽

Hepatoma Cell ◽

Hepatoma Cells ◽

Mitotic Entry ◽

Hepatoma Cell Lines ◽

Jnk Activation ◽

Cells Death ◽

Chang Liver

Hepatocellular carcinoma (HCC) generally shows chemoresistant features to anticancer agents. Paclitaxel has been clinically used in the treatment of various cancers. However, effect of paclitaxel on HCC has not been adequately addressed. Here, we found two categories of hepatoma cells in response to paclitaxel. Paclitaxel effectively decreased the cell viability of SNU475, Hep3B, and SNU387 HCC cells and Chang liver cells (death prone). In contrast, the other five hepatoma cell lines (SNU449, SNU398, SUN368, SNU354, and HepG2 cells) were resistant to paclitaxel (death reluctant). In response to paclitaxel, Bcl-2 was highly phosphorylated in death-prone cells, whereas much less Bcl-2 was phosphorylated in death-reluctant cells. Cotreatment with SP600125, an inhibitor JNK, significantly reduced the phosphorylated Bcl-2 in death-prone cells and caused a significant reduction in cell death. The reduced cell death was due to prohibition into mitotic entry as evidenced by low cyclin B1/Cdk1 kinase activity. In death-reluctant cells, inbuild-phospho-JNK levels were high but no longer activated in response to paclitaxel. We found that paclitaxel combined with caffeine or UCN-01, inhibitors of G2 DNA damage checkpoint, was able to partially overcome resistance to paclitaxel in these cells. Thus our data provide the molecular basis of paclitaxel resistance in hepatoma cells, and appropriate combination therapy may increase treatment efficacy.

Download Full-text

Increase in class 2 aldehyde dehydrogenase expression by arachidonic acid in rat hepatoma cells

Biochemical Journal ◽

10.1042/bj3570811 ◽

2001 ◽

Vol 357 (3) ◽

pp. 811-818 ◽

Cited By ~ 6

Author(s):

Rosa A. CANUTO ◽

Margherita FERRO ◽

Raffaella A. SALVO ◽

Anna M. BASSI ◽

Antonella TROMBETTA ◽

...

Keyword(s):

Lipid Peroxidation ◽

Arachidonic Acid ◽

Cell Growth ◽

Cell Lines ◽

Aldehyde Dehydrogenase ◽

Hepatoma Cell ◽

Hepatoma Cells ◽

Hepatoma Cell Lines ◽

Rat Hepatoma ◽

Rat Hepatoma Cells

Aldehyde dehydrogenase (ALDH) is a family of several isoenzymes important in cell defence against both exogenous and endogenous aldehydes. Compared with normal hepatocytes, in rat hepatoma cells the following changes in the expression of ALDH occur: cytosolic class 3 ALDH expression appears and mitochondrial class 2 ALDH decreases. In parallel with these changes, a decrease in the polyunsaturated fatty acid content in membrane phospholipids occurs. In the present study we demonstrated that restoring the levels of arachidonic acid in 7777 and JM2 rat hepatoma cell lines to those seen in hepatocytes decreases hepatoma cell growth, and increases class 2 ALDH activity. This latter effect appears to be due to an increased gene transcription of class 2 ALDH. To account for this increase, we examined whether peroxisome-proliferator-activated receptors (PPARs) or lipid peroxidation were involved. We demonstrated a stimulation of PPAR expression, which is different in the two hepatoma cell lines: in the 7777 cell line, there was an increase in PPARα expression, whereas PPARγ expression increased in JM2 cells. We also found increased lipid peroxidation, but this increase became evident at a later stage when class 2 ALDH expression had already increased. In conclusion, arachidonic acid added to the culture medium of hepatoma cell lines is able to partially restore the normal phenotype of class 2 ALDH, in addition to a decrease in cell growth.

Download Full-text