ERK2-Pyruvate Kinase Axis Permits Phorbol 12-Myristate 13-Acetate-induced Megakaryocyte Differentiation in K562 Cells

Chaman, Noor; Iqbal, Mohammad Askandar; Siddiqui, Farid Ahmad; Prakasam, Gopinath; Bamezai, R.

doi:10.1074/jbc.m115.657411

Cited by 16 publications

(11 citation statements)

References 48 publications

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“…However, in this study, we only found that PKM2 is highly expressed in moderately differentiated HCC (Figure 7E, 7F; Table 2). This is somewhat in line with two previous findings: one is that PKM2 expression is upregulated in PMA-induced megakaryocytic differentiation and that PKM2 silencing inhibits its differentiation in leukemia [36]. The other is the interaction of PKM2 with Oct4, a major regulator of self-renewal and differentiation in stem cells, decreases the transcriptional activity of Oct4 for ‘stemness’ maintaining of glioma stem cells and thus induces differentiation [37].…”

Section: Discussionsupporting

confidence: 91%

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Phage display library selection of a hypoxia-binding scFv antibody for liver cancer metabolic marker discovery

et al. 2016

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Hypoxia, which is frequently observed in liver cancer and metastasis, influences tumor progression and resistance to therapy. Although hypoxia-associated biomarkers are of use in other cancers, none is recognized as a surrogate for hypoxia in liver cancer. In this study, we generated seven unique human single-chain Fv (scFv) antibodies (Abs) specific to hypoxic liver cancer cells, using normoxia-depleted vs hypoxia-selected phage library panning technology. By developing the scFv immunoprecipitation-based mass spectrometry method, the antigen that bound with one of the Abs (H103) was identified as the M2 splice isoform of pyruvate kinase (PKM2), an enzyme that is a key regulator of aerobic glycolysis in cancer cells. Increased expression of PKM2 was induced by hypoxia in liver cancer cell lines. Immunohistochemical (IHC) staining showed that PKM2 was highly expressed in moderately and well differentiated hepatocellular carcinoma (HCC) tissues with a hypovascular staining pattern. High expression of PKM2 was also localized in the perinecrotic area of intrahepatic cholangiocarcinoma (ICC) tissues. The percentage of the HCC or ICC tumor expressing PKM2 was significantly higher with more tumor necrosis, low microvessel density, and advanced stage. Moreover, the H103 scFv Ab was efficiently internalized into hypoxic liver cancer cells and could have potential for targeted drug delivery. Conclusion: our study, for the first time, developed hypoxia-specific scFv Ab H103 to liver cancer cells, and revealed that PKM2 is a promising biomarker for hypoxia in HCC and ICC tissues. These allow further exploration of this valuable Ab and PKM2 antigen for hypoxia targeting in liver cancer.

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Section: Discussionsupporting

confidence: 91%

“…We found in this study that hypoxia induces increased expression of the PKM2 protein, but not PKL or PKM1 in liver cancer cell lines (Figures 6, S3). This mirrors the early observation that PKM2 mRNA is up-regulated in hypoxia-treated hepatoma cells [36]. There are three possible mechanism underlying PKM2 overexpression in hypoxic cells.…”

Section: Discussionsupporting

confidence: 86%

Phage display library selection of a hypoxia-binding scFv antibody for liver cancer metabolic marker discovery

et al. 2016

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“…ERK1/2 is largely involved in megakaryocytic differentiation, the phosphorylation of ERK1/2 triggers megakaryocytic differentiation . It was shown that the phosphorylation of ERK1/2 (p‐ERK1/2) was increased after incubation with ee‐As 4 S 4 (Figure 4E).…”

Section: Resultsmentioning

confidence: 99%

Arsenic Sulfide Nanoformulation Induces Megakaryocytic Differentiation through Histone Deacetylase Inhibition

Jia

Wang

et al. 2019

Advanced Therapeutics

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Arsenic sulfide (As 4 S 4 ) is a mineral drug that can be administrated orally and has been applied in several Chinese medicine formulations for the treatment on some kinds of myeloid leukemia. However, mechanisms of As 4 S 4 are not demonstrated adequately due to the poor water solubility of As 4 S 4 in its original crystal form. The authors previously fabricated water dissolvable arsenic sulfide nanoformulation (ee-As 4 S 4 ), which downregulated intracellular reactive oxygen species (ROS). As ROS is closely associated with the histone deacetylase (HDAC), in this study the effect of ee-As 4 S 4 on the HDAC activity in chronic myeloid leukemia (CML) cells and whether the effect can induce subsequent megakaryocytic differentiation and underlying mechanisms are investigated. Results show that ee-As 4 S 4 inhibits HDAC activity in CML cells, resulting in the suppression of cellular respiration. The inhibition of HDAC activity also leads to the transcriptional activation of RUNX1, which induces megakaryocytic differentiation in CML cell lines, evidenced by the significantly multiple nuclei and giant nuclear structure, upregulation of CD41a and the increased phosphorylation of ERK1/2. In conclusion, ee-As 4 S 4 is able to inhibit HDAC activity, and therefore induce significant megakaryocytic differentiation in CML cells through RUNX1-dependent pathways, which suggests its clinical potential as a complementary drug option for CML patients.

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“…A). Since PMA is known to stimulate NF‐κB p65 subunit and PKM2 dimerization (Amini et al, ; Kaminski et al, ; Chaman et al, ), we assessed whether PMA could modulate the synergistic effects observed with PKM2 and p65 overexpression. The results showed that PMA alone induced HIV‐1 LTR activity by 16‐fold (Fig.…”

Section: Resultsmentioning

confidence: 99%

Non-Metabolic Role of PKM2 in Regulation of the HIV-1 LTR

et al. 2016

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Identification of cellular proteins, in addition to already known transcription factors such as NF-κB, Sp1, C-EBPβ, NFAT, ATF/CREB, and LEF-1, which interact with the HIV-1 LTR, is critical in understanding the mechanism of HIV-1 replication in monocytes/macrophages. Our studies demonstrate upregulation of pyruvate kinase isoform M2 (PKM2) expression during HIV-1SF162 infection of monocyte/macrophages and reactivation of HIV-1 in U1 cells, a macrophage model of latency. We observed that HIV-1SF162 infection of monocyte/macrophages and reactivation of HIV-1 in U1 cells by PMA resulted in increased levels of nuclear PKM2 compared to PMA-induced U937 cells. Furthermore, there was a significant increase in the nuclear dimeric form of PKM2 in the PMA-induced U1 cells in comparison to PMA-induced U937 cells. We focused on understanding the potential role of PKM2 in HIV-1 LTR transactivation. Chromatin immunoprecipitation (ChIP) analysis in PMA-activated U1 and TZM-bl cells demonstrated the interaction of PKM2 with the HIV-1 LTR. Our studies show that overexpression of PKM2 results in transactivation of HIV-1 LTR-luciferase reporter in U937, U-87 MG, and TZM-bl cells. Using various truncated constructs of the HIV-1 LTR, we mapped the region spanning −120 bp to −80 bp to be essential for PKM2-mediated transactivation. This region contains the NF-κB binding site and deletion of this site attenuated PKM2-mediated activation of HIV-1 LTR. Immunoprecipitation experiments using U1 cell lysates demonstrated a physical interaction between PKM2 and the p65 subunit of NF-κB. These observations demonstrate for the first time that PKM2 is a transcriptional co-activator of HIV-1 LTR.

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ERK2-Pyruvate Kinase Axis Permits Phorbol 12-Myristate 13-Acetate-induced Megakaryocyte Differentiation in K562 Cells

Cited by 16 publications

References 48 publications

Phage display library selection of a hypoxia-binding scFv antibody for liver cancer metabolic marker discovery

Phage display library selection of a hypoxia-binding scFv antibody for liver cancer metabolic marker discovery

Arsenic Sulfide Nanoformulation Induces Megakaryocytic Differentiation through Histone Deacetylase Inhibition

Non-Metabolic Role of PKM2 in Regulation of the HIV-1 LTR

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