Acute Monocytic Leukemic Cell Research Articles

Solasonine Suppresses the Proliferation of Acute Monocytic Leukemia Through the Activation of the AMPK/FOXO3A Axis.

Solasonine, the main active ingredient of Solanum nigrum L., has been reported to exert extensive antitumor activity. However, the antitumor effects in acute monocytic leukemia and the exact mechanisms involved are unknown. In this study, we investigated the role of solasonine on inhibiting the progression of acute monocytic leukemia. Our findings showed that solasonine inhibited the proliferation of acute monocytic leukemic cell lines (THP-1 and MV4-11) in vitro. Solasonine promoted apoptosis and induced cell cycle arrest in the G2/M phase. Analysis of RNA-seq data suggested that solasonine correlated with increased expression of genes in the AMPK/FOXO3A pathway. Inhibition of AMPK with compound C followed by treatment with solasonine showed that solasonine reduced apoptosis, caused less cell cycle arrest, and inactivated the AMPK/FOXO3A axis in THP-1 and MV4-11 cells. Solasonine also inhibited tumor growth by the activation of the AMPK/FOXO3A axis. In conclusion, solasonine inhibited the progress of acute monocytic leukemia in vitro and in vivo and triggered the apoptosis and cell cycle arrest in the G2/M phase by upregulating the AMPK/FOXO3A pathway.

Abstract B32: A specialized post-transcriptional program in chemoresistant, quiescent cancer cells

Abstract Quiescent (G0) cells are a clinically relevant fraction in cancers, which include dormant cancer stem cells, and resist clinical therapy. G0 cells reveal extensive changes in gene expression at the protein and translation levels. We previously identified that the translation mechanism is altered in G0 cancer cells. MicroRNAs, noncoding RNAs that target distinct mRNAs to alter gene expression, were found to associate with a key RNA-binding protein and enable specialized functions in G0, where they recruit noncanonical translation factors to regulate specific mRNA translation. We find that G0 leukemic cells show similar proteome and translatome to cells isolated post-chemotherapy. These data suggest that specialized post-transcriptional mechanisms in G0 leukemic cells regulate a distinct translatome to mediate chemoresistance. To understand the role of post-transcriptional regulation in chemoresistance, we compared global transcriptome, translatome and proteome profiling in chemoresistant G0 acute monocytic leukemic (AML) cells. We find that chemotherapy or G0 induction leads to DNA damage responsive ATM and stress signaling, which alter post-transcriptional and translational mechanisms. ATM and stress-activated p38 MAPK/MK2 increase AU-rich-element (ARE) bearing proinflammatory cytokine and immune gene mRNAs, by regulating a key ARE RNA binding protein and modifying canonical translation. AREs are present on 3'UTRs of tightly regulated oncogenes and cytokines, to post-transcriptionally control their expression. Both rate-limiting steps—mRNA cap recognition and tRNA recruitment—in canonical translation are altered. These signaling pathways lead to low mTOR activity in G0, which activates the cap complex inhibitor, eIF4EBP to impair canonical translation, leading to noncanonical translation of specific mRNAs with specialized cap binding and ribosome recruitment factors. In addition, stress and STAT1/interferon signaling are activated to reduce the canonical tRNA recruitment mechanism, enabling noncanonical translation of specific mRNAs. These changes permit translation of ARE bearing proinflammatory cytokine TNFa, and immune and cell-migration modulators that promote survival. Co-inhibiting p38 MAPK and TNFa that promote antiapoptosis—prior to or along with chemotherapy—decreases chemoresistance in AML cells, in vivo, and in patient samples without affecting normal cells. Our studies reveal a proinflammatory subpopulation in AML that mediates resistance, enabled by DNA damage- and stress-regulated post-transcriptional and translational mechanisms that are mediated by AU-rich-elements and a critical ARE RNA binding protein. Disrupting ARE regulation reduces TNFα and chemoresistance, revealing AREs, an important ARE RNA binding protein and noncanonical translation as regulators of chemoresistance. These studies reveal the significance of post-transcriptional regulation of proinflammatory and immune gene-mediated chemoresistance. Citation Format: Sooncheol Lee, Syed I.A. Bukhari, Samuel S. Truesdell, Swapna Kollu, Richard D. Mortensen, Myriam Boukhali, Esha Jain, Dongjun Lee, Maria Mazzola, Radhika Raheja, Adam Langenbucher, Nicholas Haradhwala, Akiko Yanagiya, Michael Lawrence, Roopali Gandhi, Ruslan Sadreyev, David Sweetser, Wilhelm Haas, Shobha Vasudevan. A specialized post-transcriptional program in chemoresistant, quiescent cancer cells [abstract]. In: Proceedings of the AACR Special Conference on Targeting PI3K/mTOR Signaling; 2018 Nov 30-Dec 8; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(10_Suppl):Abstract nr B32.

A specialized post‐transcriptional program in chemoresistant, quiescent cancer cells

Quiescent (G0) cells are a relevant fraction in cancers, which include dormant stem cells, and resist clinical therapy. G0 cells reveal changes in gene expression at the protein and translation levels. We previously identified that the translation mechanism is altered in G0 cancer cells. MicroRNAs—noncoding RNAs that target mRNAs to alter gene expression—associate with a key RNA‐binding protein, and recruit non‐canonical translation factors to mediate specific mRNA translation in G0. We find that G0 leukemic cells show similar proteome and translatome to cells isolated post‐chemotherapy. These data suggest that specialized post‐transcriptional mechanisms in G0 leukemic cells produce a distinct translatome to facilitate chemoresistance.To understand post‐transcriptional regulation in chemoresistance, we compared global transcriptome, translatome and proteome in chemoresistant G0 acute monocytic leukemic (AML) cells. We find that chemotherapy or G0 leads to DNA damage responsive ATM and stress signaling, which alter post‐transcriptional and translational mechanisms. ATM and stress activated p38 MAPK/MK2 increase AU‐rich‐element (ARE) bearing pro‐inflammatory cytokine and immune gene mRNAs, by regulating a key ARE RNA binding protein and modifying canonical translation. AREs are present on 3′UTRs of tightly regulated cytokines to post‐transcriptionally control their expression. Both rate limiting steps—tRNA recruitment and mRNA cap recognition—in canonical translation are altered. Stress signaling activates kinases that reduce canonical tRNA recruitment. This signaling also reduces mTOR activity in G0 and chemoresistant cells, which activates the cap complex inhibitor, eIF4EBP to impair canonical translation, leading to non‐canonical translation of specific mRNAs with specialized cap binding and ribosome recruitment factors.These changes permit translation of ARE bearing pro‐inflammatory cytokine TNFα, and immune modulators that promote survival. Co‐inhibiting p38 MAPK and TNFα that promote anti‐apoptosis— prior to or along with chemotherapy—decreases chemoresistance in AML cells, in vivo, and in patient samples, without affecting normal cells. Our studies reveal a pro‐inflammatory subpopulation in AML that mediates resistance, enabled by DNA damage‐ and stress‐regulated post‐transcriptional and translational mechanisms that are mediated by AU‐rich‐elements and a critical ARE RNA binding protein. Disrupting ARE regulation reduces TNFα and chemoresistance, revealing AREs, the ARE RNA binding protein and non‐canonical translation as regulators of chemoresistance. These studies reveal the importance of post‐transcriptional regulation of pro‐inflammatory genes in chemoresistance.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Laser assisted synthesis of inorganic fullerene like MoS2-Au nanohybrid and their cytotoxicity against human monocytic (THP-1) cells

Here in, we report the investigation of the immunotoxicity of gold nanoparticles decorated on inorganic fullerene like MoS2 nanostructure (IFMoS2- AuNPs) on the THP-1 immune cell line. The MoS2 nanoparticle with fullerene like nanostructure (IFMoS2) was synthesized by double pulsed laser-assisted chemical vapour deposition (LCVD) method from bulk MoS2 at the temperature of 700 °C. The MoS2 inorganic fullerene-like nanoparticles grown by vapour-solid process (VS). The surface of the IFMoS2 was decorated with gold nanoparticles (AuNPs) to develop the semiconductor biocompatibility interface. The IFMoS2 are typically with diameters 20–50 nm as observed from the electron microscopy analysis. The stability of IFMoS2-AuNPs were examined by dynamic light scattering (DLS) and zeta potential measurements. The decoration of AuNPs on IFMoS2 surface was analyzed by UV–Vis, FTIR, PXRD, FESEM, EDX with elemental mapping and HRTEM measurement. The in vitro cytotoxicity evaluation of the FMoS2-AuNPs was determined on acute monocytic leukemic cells (THP-1) in which cell viability, caspase activity (CASP -3&7/CASP -8/CASP -9) and cell- structural changes were assessed. From FESEM revealed that IFMoS2-AuNPs induced apoptosis in THP-1 cells. Further, THP-1 cell viability markedly decreased at higher IFMoS2-AuNPs concentrations (65–100 μg/ml), indicating its potential as a chemotherapeutic agent in the treatment of human monocytic leukemia.

Abstract 4443: A post-transcriptional program of chemoresistance regulators in quiescent cancer cells

Abstract Quiescent (G0) cells are a clinically relevant fraction in several cancers, which include dormant cancer stem cells, and resist clinical therapy. G0 cells reveal extensive changes in gene expression at the protein and translation levels. We previously identified that the translation mechanism is altered in G0 cancer cells. MicroRNAs, noncoding RNAs that target distinct mRNAs to alter gene expression—were found to associate with an important RNA-binding protein, and enable specialized functions in G0—where they recruit non-canonical translation factors to regulate specific mRNA translation. We find that G0 leukemic cells show similar proteome and translatome to cells isolated post-chemotherapy. These data suggest that specialized post-transcriptional and translational mechanisms in G0 leukemic cells regulate a distinct translatome to mediate chemoresistance. To understand the role of post-transcriptional and translational regulation in chemoresistance, we compared global RNA, translational and proteome profiling in chemoresistant G0 acute monocytic leukemic (AML) cells. We find that chemotherapy or G0 induction leads to DNA damage responsive ATM and stress signaling, which alter post-transcriptional and translational mechanisms. ATM and stress activated p38 MAPK/MK2 increase AU-rich-element (ARE) bearing pro-inflammatory cytokine and immune gene mRNAs by regulating a key ARE RNA binding protein, and by activating STAT1/interferon pathway to alter canonical translation. AREs are present on 3'UTRs of critical, tightly regulated oncogenes and cytokines to post-transcriptionally control their expression. These changes permit translation of ARE bearing pro-inflammatory cytokine TNFα, and immune and cell-migration modulators that promote survival. Co-inhibiting p38 MAPK and TNFα that promote anti-apoptosis—prior to or alongwith chemotherapy—decreases chemoresistance in AML cell lines, in vivo, and in patient samples, without affecting normal cells. These studies reveal a pro-inflammatory subpopulation in AML that mediates chemoresistance, enabled by DNA damage- and stress-regulated post-transcriptional and translational mechanisms that are mediated by AU-rich-elements and a critical ARE RNA binding protein. Disrupting ARE regulation reduces TNFα and chemoresistance, revealing AREs and an important ARE RNA binding protein as key regulators of inflammation-mediated chemoresistance. These studies reveal the significance of post-transcriptional regulation of inflammation/immune gene-mediated chemoresistance. Correspondence: vasudevan.shobha@mgh.harvard.edu Citation Format: Sooncheol Lee, Samuel S. Truesdell, Syed I. Bukhari, Myriam Boukhali, Dongjun Lee, Maria A. Mazzola, Radhika Raheja, Adam Langenbucher, Nicholas J. Haradhvala, Michael Lawrence, Roopali Gandhi, David A. Sweetser, Wilhelm Haas, Shobha Vasudevan. A post-transcriptional program of chemoresistance regulators in quiescent cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4443.

RNAi-mediated Silencing of CXCR4 Inhibits the Adhesion, Invasion and Tumorigenicity of Acute Monocytic Leukemic Cell Line SHI-1

To investigate the effect of CXCR4 gene on the proliferation, adhesion, invasion and tumorigenicity of a human monocytic leukemic cell line SHI-1. The lentivirus vector silencing the expression of CXCR4 was constructed for infection of SHI-1 cells silencing expression of CXCR4 in SHI-1 cells. The expression of CXCR4, MMP-2 and MMP-9 was detected by real time PCR. The expression of CXCR4 on membrane of SHI-1 cells was detected by flow cytometry. The SHI-1 cell proliferation ability was detected by MTT. The co-culture system of the leukemia cells and bone marrow stromal cells was used to detect the adhesion and migration ability of leukemia cells. SHI-1 cells were inoculated subcutaneously in nude mice to investigate the growth ability in vivo. After SHI-1 cells were infected by lentivirus silencing expression of CXCR4, the expression of CXCR4 mRNA in SHI-1 CXCR-4i cells decreased by 76% as compared with expression of SHI-1/NC of negative control virus, the expression of CXCR4 on membrane of SHI-1/CXCR4i obviously downregulated; the expression of MMP-2 and MMP-9 in SHI-1/CXCRi cells also declined by 63% and 62% respectively; the proliferation ability of SHI-1/CXCR4i in vitro did not obviously changed, but the adhesion and trans-matrigel invasion ability significantly decreased, the SHI-1/CXCR4i cells could not form neoplasm subcutaneously in mice, but the SHI-1 and SHI-1/NC cells could form neoplasm subcutaneously in mice, and there was no significant difference in volumn of neoplasm mass. The silencing expression of CXCR4 can decline the adhesion and migration ability of SHI-1 cells, and can completely suppress the formation of neoplasm subcutaneously, so the CXCR4 may serve as a target for treating leukemia.

Hypertonic saline downregulate the production level of lipopolysaccharide-induced migration inhibitory factor in THP-1 cells

PurposeMacrophage migration inhibitory factor (MIF) may serve as a general marker for systemic inflammation in septic and nonseptic acute critical illness. Additionally, our previous experiment has demonstrated that immunosuppressant Prostaglandin E2 (PGE2) lowered MIF levels and inhibited T-cells proliferation when compared to control levels. The addition of hypertonic saline (HTS) increased MIF production as compared with PGE2-stimulated T-cells in concordance with restore PGE2-suppressed T-cells proliferation. Generally, HTS has been well known for its anti-inflammatory effect so far. Therefore, the experiments were conducted to evaluate MIF after stimulating lipopolysaccharide (LPS) either in the presence or absence of HTS in monocyte, in response to early phase injury.MethodsHuman acute monocytic leukemic cell line (THP-1) cells were cultured in RPMI media, to a final concentration of 1 × 106 cells/mL. The effect of HTS on LPS-induced MIF was evaluated in monocyte with 1 µg/mL LPS. HTS at 10, 20 or 40 mmol/L above isotonicity was added. MIF concentrations of the supernatant were determined by enzyme-linked immunosorbent assay, and cell lysates were used for Western blots analysis to determine the MIF expression.ResultsMIF concentrations in the cell supernatant increased in LPS-induced cells compared to control cells. Also, levels of MIF protein expression were higher in LPS stimulating cells. However, the addition of HTS to LPS stimulated cell restored MIF concentrations and MIF expression.ConclusionThe role of HTS in maintaining physiological balance in human beings, at least in part, should be mediated through the MIF pathway.

Effect of siRNA targeting Ets2 gene on chemosensitization of human acute monocytic leukemic cell line SHI-1

Objective This study was to observe the levels of Ets2 mRNA expression in leukemia patients and investigate the effect of small interfering RNA (siRNA) targeting Ets2 gene on sensibility of human acute monocytic leukemic cell line SHI-1 cells to etoposide (VP-16). SHI-1 cells to etoposide (VP-16).

Preliminary Study of Inducing Differentiation and Apoptosis in a Novel Human Acute Monocytic Leukemic Cell Line SHI-1.

ObjectiveTo explore the inducing differentiation and apoptosis in a novel human acute monocytic leukemic cell line SHI-1.MethodsCell morphological analysis, NBT reduction test, the expression of CD11b and CD14, cell cycle analysis and binding of annexinV tested by FCM, DNA electrophoresis were performed to evaluate whether tributyrin (TB), arsenic trioxide (As2O3), and 12-O-tetradecanoyl-phorbol-13 acetate (TPA) could induce the differentiation and apoptosis in the SHI-1 cell line.ResultsThe results of morphological analysis, NBT reduction test, and FCM assay showed that TB could induce differentiation of SHI-1 cells. The SHI-1 cell line presented typical apoptotic evidences including morphology features, DNA ladders and the increased apoptotic rate determined by FCM. Moreover, the SHI-1 cell line presented typical apoptotic morphology after being treated with As2O3. The apoptotic rate of the SHI-1 cell line treated with As2O3 was increased determined by FCM. The expression level of CD14 was upregulated while that of CD11b was unchanged in the SHI-1 cell line. Most of the SHI-1 cells adhered to the flask after being treated with TPA for 24 hours.ConclusionsTB may produce dual effects of differentiation and apoptosis. The cell line could be induced to differentiation partially or apoptosis after being treated with 0.5μM, 1.0μM, and 2.0μM As2O3. TPA could induce differentiation and adhesion of the SHI-1 cell line.

The chromosome translocation t(7;11)(p15;p15) in acute myeloid leukemia results in fusion of the NUP98 gene with a HOXA cluster gene, HOXA13, but not HOXA9.

The nucleoporin gene NUP98 has been reported to be fused to 9 partner genes in hematologic malignancies with 11p15 translocations. The NUP98-HOXA9 fusion gene has been identified in acute myeloid leukemia (AML) and chronic myelogenous leukemia with t(7;11)(p15;p15). We report here a novel NUP98 partner gene, HOXA13, in a patient with de novo AML having t(7;11)(p15;p15). The HOXA13 gene is part of the HOXA cluster genes and contains 2 exons, encoding a protein of 338 amino acids with a homeodomain. The NUP98-HOXA13 fusion protein consists of the N-terminal phenylalanine-glycine repeat motif of NUP98 and the C-terminal homeodomain of HOXA13, similar to the NUP98-HOXA9 fusion protein. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis in various leukemic cell lines showed that the HOXA13 gene was expressed significantly more frequently in acute monocytic leukemic cell lines than in other leukemic cell lines (P = 0.039). HOXA13 and three HOXA cluster genes (A9, A10, A11) located at the 5' end of the HOXA9 gene were frequently expressed in myeloid leukemic cell lines. Our results revealed that t(7;11)(p15;p15) was not a single chromosomal abnormality at the molecular level. The protein encoded by the NUP98-HOXA13 fusion gene is similar to that encoded by NUP98-HOXA9, and the expression pattern of the HOXA13 gene in leukemic cell lines is similar to that of the HOXA9 gene, suggesting that the NUP98-HOXA13 fusion protein may play a role in leukemogenesis through a mechanism similar to that of the NUP98-HOXA9 fusion protein.

Release of monocyte chemoattractants by polymorphonuclear leukocytes stimulated by their adhesion to stratum corneum opsonized via complement activation, measured with a human acute monocytic leukemic cell line, THP-1.

Stratum corneum (SC) exposed to living tissues, induces inflammation characterized by the formation of mixed cell granulomas consisting of infiltrative polymorphonuclear leukocytes (PMNs) and monocytes/macrophages. In this study, to clarify the mechanism for the later monocyte accumulation in SC-induced granulomas, we evaluated monocyte chemotactic activity induced by PMNs treated with serum-opsonized SC by using a human acute monocytic leukemic cell line, THP-1. When the supernatant was obtained from a PMN suspension cultured with opsonized plantar SC, higher THP-1 chemotactic activity was detected as compared with that cultured with non-opsonized SC. Although some concentrations of the chemokines, MIP-1alpha and MIP-1beta, were detected in supernatants obtained from the PMN suspensions cultured with plantar SC than in the control suspensions of PMN alone, their production by PMN was not influenced by the opsonization procedure. In contrast, MCP-1 was found to be secreted from PMN suspensions constitutively, showing no correlation to this THP-1 chemotactic activity. Moreover, HPLC analysis of PMN suspensions indicated that factors with far higher molecular weight values than these chemokines are involved in the chemotaxis of THP-1 cells.

Thymidine kinase isoenzymes in human acute and chronic lymphatic leukemia

The dominating thymidine kinase isoenzyme was examined in mononuclear leucocytes from two patients, one with acute and one with chronic lymphatic leukemia. The two isoenzymes exhibited Michaelis-Menten substrate kinetics with ATP and cooperative inhibition kinetics with dTTP. The substrate kinetics with thymidine were different. According to the enzymatic properties the isoenzymes from the acute and chronic lymphatic cells were designated TK 3 and TK 4, respectively. Comparison with the isoenzymes in normal lymphocytes (TK 1, TK 2) and in acute monocytic leukemic cells (TK 3, TK 4) indicated the existence of three thymidine kinase isoenzymes in human leukemic cells which differed from the two isoenzymes in normal human lymphocytes.