Targeting PML-RARαand Oncogenic Signaling Pathways by Chinese Herbal MixtureTien-HsienLiquid in Acute Promyelocytic Leukemia NB4 Cells

Tien-HsienLiquid (THL) is a Chinese herbal mixture that has been used worldwide as complementary treatment for cancer patients in the past decade. Recently, THL has been shown to induce apoptosis in various types of solid tumor cellsin vitro. However, the underlying molecular mechanisms have not yet been well elucidated. In this study, we explored the effects of THL on acute promyelocytic leukemia (APL) NB4 cells, which could be effectively treated by some traditional Chinese remedies containing arsenic trioxide. The results showed THL could induce G2/M arrest and apoptosis in NB4 cells. Accordingly, the decrease of cyclin A and B1 were observed in THL-treated cells. The THL-induced apoptosis was accompanied with caspase-3 activation and decrease of PML-RARαfusion protein. Moreover, DNA methyltransferase 1 and oncogenic signaling pathways such as Akt/mTOR, Stat3 and ERK were also down-regulated by THL. By using ethyl acetate extraction and silica gel chromatography, an active fraction of THL named as EAS5 was isolated. At about 0.5–1% of the dose of THL, EAS5 appeared to have most of THL-induced multiple molecular targeting effects in NB4 cells. Based on the findings of these multi-targeting effects, THL might be regarding as a complementary and alternative therapeutic agent for refractory APL.

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Arsenic Trioxide Promotes Histone H3 Phosphoacetylation at the Chromatin ofCASPASE-10in Acute Promyelocytic Leukemia Cells

Journal of Biological Chemistry ◽

10.1074/jbc.m207836200 ◽

2002 ◽

Vol 277 (51) ◽

pp. 49504-49510 ◽

Cited By ~ 73

Author(s):

Ji Li ◽

Peili Chen ◽

Natasha Sinogeeva ◽

Myriam Gorospe ◽

Robert P. Wersto ◽

...

Keyword(s):

Arsenic Trioxide ◽

Acute Promyelocytic Leukemia ◽

Molecular Mechanisms ◽

Expression Profiles ◽

Histone H3 ◽

Gene Expression Profiles ◽

Promyelocytic Leukemia ◽

Therapeutic Effects ◽

Acid Therapy ◽

Nb4 Cells

Arsenic trioxide (As2O3) is highly effective for the treatment of acute promyelocytic leukemia, even in patients who are unresponsive to all-trans-retinoic acid therapy. As2O3is believed to function primarily by promoting apoptosis, but the underlying molecular mechanisms remain largely unknown. In this report, using cDNA arrays, we have examined the changes in gene expression profiles triggered by clinically achievable doses of As2O3in acute promyelocytic leukemia NB4 cells.CASPASE-10expression was found to be potently induced by As2O3. Accordingly, caspase-10 activity also substantially increased in response to As2O3treatment. A selective inhibitor of caspase-10, Z-AEVD-FMK, effectively blocked caspase-3 activation and significantly attenuated As2O3-triggered apoptosis. Interestingly, the treatment of NB4 cells with As2O3markedly increased histone H3 phosphorylation at serine 10, an event that is associated with acetylation of the lysine 14 residue. Chromatin immunoprecipitation assays revealed that As2O3potently enhances histone H3 phosphoacetylation at theCASPASE-10locus. These results suggest that the effect of As2O3on histone H3 phosphoacetylation at theCASPASE-10gene may play an important role in the induction of apoptosis and thus contribute to its therapeutic effects on acute promyelocytic leukemia.

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Retinoid-induced differentiation of acute promyelocytic leukemia involves PML-RARalpha-mediated increase of type II transglutaminase

Blood ◽

10.1182/blood.v87.5.1939.bloodjournal8751939 ◽

1996 ◽

Vol 87 (5) ◽

pp. 1939-1950 ◽

Cited By ~ 7

Author(s):

L Benedetti ◽

F Grignani ◽

BM Scicchitano ◽

AM Jetten ◽

D Diverio ◽

...

Keyword(s):

Cell Line ◽

Signaling Pathway ◽

Acute Promyelocytic Leukemia ◽

Molecular Mechanisms ◽

Growth Arrest ◽

Promyelocytic Leukemia ◽

Leukemic Cells ◽

Type Ii ◽

Granulocytic Differentiation ◽

Nb4 Cells

All-trans retinoic acid (t-RA) administration leads to complete remission in acute promyelocytic leukemia (APL) patients by inducing growth arrest and differentiation of the leukemic clone. In the present study, we show that t-RA treatment dramatically induced type II transglutaminase (type II TGase) expression in cells carrying the t(15;17) translocation and expressing the PML-RARalpha product such as the APL-derived NB4 cell line and fresh leukemic cells from APL patients. This induction correlated with t-RA-induced growth arrest, granulocytic differentiation, and upregulation of the leukocyte adherence receptor beta subunit (CD18) gene expression. The increase in type II TGase was not abolished by cycloheximide treatment, suggesting that synthesis of a protein intermediate was not required for the induction. t-RA did not significantly alter the rate of growth arrest and did not stimulate differentiation and type II TGase activity in NB4.306 cells, a t-RA-resistant subclone of the NB4 cell line, or in leukemic cells derived from two patients morphologically defined as APL but lacking the t(15;17). However, in NB4.306 cells, t-RA treatment was able to increase CD18 mRNA expression in a manner similar to NB4 cells. The molecular mechanisms involved in the induction of these genes were investigated. In NB4 cells, using novel receptor-selective ligands such as 9-cis-RA, TTNPB, AM580, and SR11217, we found that RAR- and RARalpha- selective retinoids were able to induce growth arrest, granulocytic differentiation, and type II TGase, whereas the RXR-selective retinoid SR11217 was inactive. Moreover, an RAR alpha-antagonist completely inhibited the expression of type II TGase and CD18 induced by these selective retinoids in NB4 cells. In NB4.306 cells, an RARalpha- dependent signaling pathway was found involved in the modulation of CD18 expression. In addition, expression of the PML-RARalpha gene in myeloid U937 precursor cells resulted in the ability of these cells to induce type II TGase in response to t-RA. On the basis of these results we hypothesize a specific involvement of a signaling pathway involving PML-RAR alpha for the induction of growth arrest, granulocytic differentiation, and type II TGase by retinoids in APL cells.

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Transcriptional and Metabolic Dissection of ATRA-Induced Granulocytic Differentiation in NB4 Acute Promyelocytic Leukemia Cells

Cells ◽

10.3390/cells9112423 ◽

2020 ◽

Vol 9 (11) ◽

pp. 2423

Author(s):

Jacopo Albanesi ◽

Nelida Ines Noguera ◽

Cristina Banella ◽

Tommaso Colangelo ◽

Elisabetta De Marinis ◽

...

Keyword(s):

Acute Promyelocytic Leukemia ◽

Molecular Mechanisms ◽

Immune Cell ◽

Aerobic Glycolysis ◽

Therapy Resistance ◽

Therapeutic Strategies ◽

Promyelocytic Leukemia ◽

Metabolic Reprogramming ◽

Granulocytic Differentiation ◽

Nb4 Cells

Acute promyelocytic leukemia (APL) is a hematological disease characterized by a balanced reciprocal translocation that leads to the synthesis of the oncogenic fusion protein PML-RARα. APL is mainly managed by a differentiation therapy based on the administration of all-trans retinoic acid (ATRA) and arsenic trioxide (ATO). However, therapy resistance, differentiation syndrome, and relapses require the development of new low-toxicity therapies based on the induction of blasts differentiation. In keeping with this, we reasoned that a better understanding of the molecular mechanisms pivotal for ATRA-driven differentiation could definitely bolster the identification of new therapeutic strategies in APL patients. We thus performed an in-depth high-throughput transcriptional profile analysis and metabolic characterization of a well-established APL experimental model based on NB4 cells that represent an unevaluable tool to dissect the complex mechanism associated with ATRA-induced granulocytic differentiation. Pathway-reconstruction analysis using genome-wide transcriptional data has allowed us to identify the activation/inhibition of several cancer signaling pathways (e.g., inflammation, immune cell response, DNA repair, and cell proliferation) and master regulators (e.g., transcription factors, epigenetic regulators, and ligand-dependent nuclear receptors). Furthermore, we provide evidence of the regulation of a considerable set of metabolic genes involved in cancer metabolic reprogramming. Consistently, we found that ATRA treatment of NB4 cells drives the activation of aerobic glycolysis pathway and the reduction of OXPHOS-dependent ATP production. Overall, this study represents an important resource in understanding the molecular “portfolio” pivotal for APL differentiation, which can be explored for developing new therapeutic strategies.

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In vitro studies on cellular and molecular mechanisms of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia: As2O3 induces NB4 cell apoptosis with downregulation of Bcl-2 expression and modulation of PML-RAR alpha/PML proteins

Blood ◽

10.1182/blood.v88.3.1052.bloodjournal8831052 ◽

1996 ◽

Vol 88 (3) ◽

pp. 1052-1061 ◽

Cited By ~ 24

Author(s):

GQ Chen ◽

J Zhu ◽

XG Shi ◽

JH Ni ◽

HJ Zhong ◽

...

Keyword(s):

Gene Expression ◽

Arsenic Trioxide ◽

Acute Promyelocytic Leukemia ◽

Cell Apoptosis ◽

Molecular Mechanisms ◽

Nuclear Dna ◽

Staining Pattern ◽

Promyelocytic Leukemia ◽

Nb4 Cells

It has been shown recently in China that arsenic trioxide (As2O3) is a very effective treatment for acute promyelocytic leukemia (APL). APL patients resistant to all-trans retinoic acid (ATRA) and conventional chemotherapy can still respond to AS2O3. In this study, we addressed the possible cellular and molecular mechanisms of this treatment by using NB4 cells as a model. The results show that: (1) As2O3 triggers relatively specific NB4 cell apoptosis at micromolar concentration, as proved by morphology, histogramic related nuclear DNA contents, and DNA gel eletrophoresis. (2) As2O3 does not influence bax, bcl-x, c-myc, and p53 gene expression, but downregulates bcl-2 gene expression at both mRNA and protein levels. (3) As2O3 induces a significant modulation of the PML staining pattern in NB4 cells and HL-60 cells. The micropunctates characteristic of PML-RAR alpha in NB4 cells dissappear after treatment with As2O3, whereas a diffuse PML staining occurs in the perinuclear cytoplasmic region. In addition, a low percentage of untreated NB4 cells exhibits an accumulation of PML positive particles in a compartment of cytoplasm. The percentage of these cells can be significantly increased after As2O3 treatment. A similar PML staining pattern is observed in apoptotic cells. (4) ATRA pretreatment does not influence As2O3-induced apoptosis. These results suggest that induction of cell apoptosis can be one of the mechanisms of the therapeutic effect of As2O3. Moreover, this apoptosis induction occurs independently of the retinoid pathway and may be mediated, at least partly, through the modulation of bcl-2, as well as PML-RAR alpha and/ or PML proteins.

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Retinoid-induced differentiation of acute promyelocytic leukemia involves PML-RARalpha-mediated increase of type II transglutaminase

Blood ◽

10.1182/blood.v87.5.1939.1939 ◽

1996 ◽

Vol 87 (5) ◽

pp. 1939-1950 ◽

Cited By ~ 61

Author(s):

L Benedetti ◽

F Grignani ◽

BM Scicchitano ◽

AM Jetten ◽

D Diverio ◽

...

Keyword(s):

Cell Line ◽

Signaling Pathway ◽

Acute Promyelocytic Leukemia ◽

Molecular Mechanisms ◽

Growth Arrest ◽

Promyelocytic Leukemia ◽

Leukemic Cells ◽

Type Ii ◽

Granulocytic Differentiation ◽

Nb4 Cells

Abstract All-trans retinoic acid (t-RA) administration leads to complete remission in acute promyelocytic leukemia (APL) patients by inducing growth arrest and differentiation of the leukemic clone. In the present study, we show that t-RA treatment dramatically induced type II transglutaminase (type II TGase) expression in cells carrying the t(15;17) translocation and expressing the PML-RARalpha product such as the APL-derived NB4 cell line and fresh leukemic cells from APL patients. This induction correlated with t-RA-induced growth arrest, granulocytic differentiation, and upregulation of the leukocyte adherence receptor beta subunit (CD18) gene expression. The increase in type II TGase was not abolished by cycloheximide treatment, suggesting that synthesis of a protein intermediate was not required for the induction. t-RA did not significantly alter the rate of growth arrest and did not stimulate differentiation and type II TGase activity in NB4.306 cells, a t-RA-resistant subclone of the NB4 cell line, or in leukemic cells derived from two patients morphologically defined as APL but lacking the t(15;17). However, in NB4.306 cells, t-RA treatment was able to increase CD18 mRNA expression in a manner similar to NB4 cells. The molecular mechanisms involved in the induction of these genes were investigated. In NB4 cells, using novel receptor-selective ligands such as 9-cis-RA, TTNPB, AM580, and SR11217, we found that RAR- and RARalpha- selective retinoids were able to induce growth arrest, granulocytic differentiation, and type II TGase, whereas the RXR-selective retinoid SR11217 was inactive. Moreover, an RAR alpha-antagonist completely inhibited the expression of type II TGase and CD18 induced by these selective retinoids in NB4 cells. In NB4.306 cells, an RARalpha- dependent signaling pathway was found involved in the modulation of CD18 expression. In addition, expression of the PML-RARalpha gene in myeloid U937 precursor cells resulted in the ability of these cells to induce type II TGase in response to t-RA. On the basis of these results we hypothesize a specific involvement of a signaling pathway involving PML-RAR alpha for the induction of growth arrest, granulocytic differentiation, and type II TGase by retinoids in APL cells.

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In vitro studies on cellular and molecular mechanisms of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia: As2O3 induces NB4 cell apoptosis with downregulation of Bcl-2 expression and modulation of PML-RAR alpha/PML proteins

Blood ◽

10.1182/blood.v88.3.1052.1052 ◽

1996 ◽

Vol 88 (3) ◽

pp. 1052-1061 ◽

Cited By ~ 586

Author(s):

GQ Chen ◽

J Zhu ◽

XG Shi ◽

JH Ni ◽

HJ Zhong ◽

...

Keyword(s):

Gene Expression ◽

Arsenic Trioxide ◽

Acute Promyelocytic Leukemia ◽

Cell Apoptosis ◽

Molecular Mechanisms ◽

Nuclear Dna ◽

Staining Pattern ◽

Promyelocytic Leukemia ◽

Nb4 Cells

Abstract It has been shown recently in China that arsenic trioxide (As2O3) is a very effective treatment for acute promyelocytic leukemia (APL). APL patients resistant to all-trans retinoic acid (ATRA) and conventional chemotherapy can still respond to AS2O3. In this study, we addressed the possible cellular and molecular mechanisms of this treatment by using NB4 cells as a model. The results show that: (1) As2O3 triggers relatively specific NB4 cell apoptosis at micromolar concentration, as proved by morphology, histogramic related nuclear DNA contents, and DNA gel eletrophoresis. (2) As2O3 does not influence bax, bcl-x, c-myc, and p53 gene expression, but downregulates bcl-2 gene expression at both mRNA and protein levels. (3) As2O3 induces a significant modulation of the PML staining pattern in NB4 cells and HL-60 cells. The micropunctates characteristic of PML-RAR alpha in NB4 cells dissappear after treatment with As2O3, whereas a diffuse PML staining occurs in the perinuclear cytoplasmic region. In addition, a low percentage of untreated NB4 cells exhibits an accumulation of PML positive particles in a compartment of cytoplasm. The percentage of these cells can be significantly increased after As2O3 treatment. A similar PML staining pattern is observed in apoptotic cells. (4) ATRA pretreatment does not influence As2O3-induced apoptosis. These results suggest that induction of cell apoptosis can be one of the mechanisms of the therapeutic effect of As2O3. Moreover, this apoptosis induction occurs independently of the retinoid pathway and may be mediated, at least partly, through the modulation of bcl-2, as well as PML-RAR alpha and/ or PML proteins.

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Effect of Cassiopea andromeda Venom on P15INK4b, P21 WAF1/CIP1, P53, DNA methyltransferase 1, and Bcl-2 Genes Expression, Apoptosis Induction, and Cell Growth Inhibition in Acute Promyelocytic Leukemia NB4 Cell line

Iranian Journal of Pediatric Hematology & Oncology ◽

10.18502/ijpho.v11i2.5842 ◽

2021 ◽

Author(s):

Reza Dehghani ◽

Narges Obeidi ◽

Gholamhossein Mohebbi ◽

Ali Amrooni ◽

Zahra Derakhshan

Keyword(s):

Cell Growth ◽

Growth Inhibition ◽

Cell Line ◽

Acute Promyelocytic Leukemia ◽

Dna Methyltransferase ◽

Promyelocytic Leukemia ◽

Tetrazolium Salt ◽

Nb4 Cells ◽

P21 Waf1

Background: One of the acute hematologic malignancies is acute promyelocytic leukemia (APL) that resulted in translocation of chromosomes 15 and 17, t (15; 17), and cessation in the maturation of myeloid cell line, and ultimate aggregation of neoplastic promyelocytes. Regarding that appetence of using herbal and marine medicine studies is increasing, and on the other hand, the features of Cassiopea andromeda Venom remained unclear; this study was conducted to determine its effects on NB4 cells as a model for APL. Materials and Methods: In this experimental study, the cells were treated with C. andromeda Venom concentrations at different periods and times. Growth inhibition and toxic effects of C. andromeda Venom were evaluated through methyl thiazole tetrazolium salt reduction (MTT test). The flow cytometry analysis was carried out using 7AAD and Annexin V stains for evaluating this venom’s effect on apoptotic pathways. Besides, Real-Time polymerase chain reaction was performed to evaluate the relative gene expression. Results: C. andromeda Venom inhibited the growth of NB4 cells as correlated with concentration and time. Cell growth was inhibited by 49.1%, after 24 hours of treating NB4 cells with 1000µg/mL C. andromeda Venom. This venom increased the apoptotic process, which was then verified by 7AAD/AnnexinV staining. The fold change of p15INK4b, p21 WAF1/CIP1, P53, DNMT1, and Bcl-2 genes in the NB4 cell line were 144, 2.78, 1.75, 15.24, and 0.33, respectively, which meant that the expression level of p15INK4b, p21 WAF1/CIP1, P53, and DNMT1 were increased by 14400%, 278%, 175%, and 1524%, respectively and the expression of Bcl-2 was decreased by 67%. Conclusion: Considering the inhibitory property of C. andromeda Venom, the authors recommended it as a part of combinational medication for treating APL in animal trials and for other leukemias’ in vitro studies.

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MALAT1 as a Versatile Regulator of Cancer: Overview of the updates from Predatory role as Competitive Endogenous RNA to Mechanistic In-sights

Current Cancer Drug Targets ◽

10.2174/1568009620999200730183110 ◽

2020 ◽

Vol 20 ◽

Author(s):

Ammad Ahmad Farooqi ◽

Evangelia Legaki ◽

Maria Gazouli ◽

Silvia Rinaldi ◽

Rossana Berardi

Keyword(s):

Molecular Biology ◽

Detailed Analysis ◽

Signaling Pathways ◽

Individualized Medicine ◽

Signaling Cascades ◽

Oncogenic Signaling ◽

Non Coding Rnas ◽

Oncogenic Signaling Pathways ◽

Competitive Endogenous Rna

: Central dogma of molecular biology has remained cornerstone of classical molecular biology but serendipitous discovery of microRNAs (miRNAs) in nematodes paradigmatically shifted our current understanding of the intricate mech-anisms which occur during transitions from transcription to translation. Discovery of miRNA captured tremendous attention and appreciation and we had witnessed an explosion in the field of non-coding RNAs. Ground-breaking discoveries in the field of non-coding RNAs have helped in better characterization of microRNAs and long non-coding RNAs (LncRNAs). There is an ever-increasing list of miRNA targets which are regulated by MALAT1 to stimulate or repress expression of tar-get genes. However, in this review our main focus is to summarize mechanistic insights related to MALAT1-mediated regu-lation of oncogenic signaling pathways. We have discussed how MALAT1 modulated TGF/SMAD and Hippo pathways in various cancers. We have also comprehensively summarized how JAK/STAT and Wnt/β-catenin pathways stimulated MALAT1 expression and consequentially how MALAT1 potentiated these signaling cascades to promote cancer. MALAT1 research has undergone substantial broadening however, there is still a need to identify additional mechanisms. MALAT1 is involved in multi-layered regulation of multiple transduction cascades and detailed analysis of different pathways will be helpful in getting a step closer to individualized medicine.

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The m6A RNA methylation regulates oncogenic signaling pathways driving cell malignant transformation and carcinogenesis

Molecular Cancer ◽

10.1186/s12943-021-01356-0 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Mohammad Burhan Uddin ◽

Zhishan Wang ◽

Chengfeng Yang

Keyword(s):

Signaling Pathways ◽

Malignant Transformation ◽

Noncoding Rna ◽

Rna Modification ◽

Oncogenic Signaling ◽

Aberrant Expression ◽

Rna Methylation ◽

Rna Molecules ◽

M6a Rna Methylation ◽

Oncogenic Signaling Pathways

AbstractThe m6A RNA methylation is the most prevalent internal modification in mammalian mRNAs which plays critical biological roles by regulating vital cellular processes. Dysregulations of the m6A modification due to aberrant expression of its regulatory proteins are frequently observed in many pathological conditions, particularly in cancer. Normal cells undergo malignant transformation via activation or modulation of different oncogenic signaling pathways through complex mechanisms. Accumulating evidence showing regulation of oncogenic signaling pathways at the epitranscriptomic level has added an extra layer of the complexity. In particular, recent studies demonstrated that, in many types of cancers various oncogenic signaling pathways are modulated by the m6A modification in the target mRNAs as well as noncoding RNA transcripts. m6A modifications in these RNA molecules control their fate and metabolism by regulating their stability, translation or subcellular localizations. In this review we discussed recent exciting studies on oncogenic signaling pathways that are modulated by the m6A RNA modification and/or their regulators in cancer and provided perspectives for further studies. The regulation of oncogenic signaling pathways by the m6A modification and its regulators also render them as potential druggable targets for the treatment of cancer.

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