Quantitative RT-PCR analysis of sphingolipid metabolic enzymes in acute leukemia and myelodysplastic syndromes

Sobue, Satoshi; Iwasaki, Takashi; Sugisaki, Chiho; Nagata, Kazuya; Kikuchi, Ryosuke; Murakami, Masashi; Takagi, Akira; Kojima, Tetsuhito; Banno, Yoshiko; Akao, Yukihiro; Nozawa, Yoshinori; Kannagi, Reiji; Suzuki, Motoshi; Abe, Akihiro; Naoe, Tomoki; Murate, Takashi

doi:10.1038/sj.leu.2404386

Cited by 58 publications

(54 citation statements)

References 6 publications

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“…Our findings, which demonstrate that SphK1 inhibition restores the susceptibility of the resistant cell lines to IM, are consistent with the idea that S1P induces cell survival mechanisms under the resistant status. Our results are also consistent with previous data showing a role for SphK1 in the regulation of drug-induced apoptosis via alteration of the ceramide/S1P balance and in the onset of chemotherapy resistance in leukemia [43] and in cells with multidrugresistant phenotype [12,35,36,44].…”

Section: Discussionsupporting

confidence: 93%

Sphingosine kinase 1 overexpression is regulated by signaling through PI3K, AKT2, and mTOR in imatinib-resistant chronic myeloid leukemia cells

Marfè

Stefano

Gambacurta

et al. 2011

Experimental Hematology

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Objective. As a better understanding of the molecular basis of carcinogenesis has emerged, oncogene-specific cell-signaling pathways have been successfully targeted to treat human malignances. Despite impressive advances in oncogene-directed therapeutics, genetic instability in cancer cells often manifest acquired resistance. This is particularly noted in the use of tyrosine kinase inhibitors therapies and not more evident than for chronic myeloid leukemia. Therefore, it is of great importance to understand the molecular mechanisms affecting cancer cell sensitivity and resistance to tyrosine kinase inhibitors. Materials and Methods. In this study, we used continuous exposure to stepwise increasing concentrations of imatinib (0.6L1 mM) to select imatinib-resistant K562 cells. Results. Expression of BCR-ABL increased both at RNA and protein levels in imatinib-resistant cell lines. Furthermore, expression levels of sphingosine kinase 1 (SphK1) were increased significantly in resistant cells, channeling sphingoid bases to the SphK1 pathway and activating sphingosine-1-phosphateLdependent tyrosine phosphorylation pathways that include the adaptor protein Crk. The partial inhibition of SphK1 activity by N,N-dimethylsphingosine or expression by small interfering RNA increased sensitivity to imatinib-induced apoptosis in resistant cells and returned BCR-ABL to baseline levels. To determine the resistance mechanism-induced SphK1 upregulation, we used pharmacological inhibitors of the phosphoinositide 3-kinase/ AKT/mammalian target of rapamycin signaling pathway and observed robust downmodulation of SphK1 expression and activity when AKT2, but not AKT1 or AKT3, was suppressed.Conclusions. These results demonstrate that SphK1 is upregulated in imatinib-resistant K562 cells by a pathway contingent on a phosphoinositide 3-kinase/AKT2/mammalian target of rapamycin signaling pathway. We propose that SphK1 plays an important role in development of acquired resistance to imatinib in chronic myeloid leukemia cell lines. Ó

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Section: Discussionsupporting

confidence: 93%

Sphingosine kinase 1 overexpression is regulated by signaling through PI3K, AKT2, and mTOR in imatinib-resistant chronic myeloid leukemia cells

Marfè

Stefano

Gambacurta

et al. 2011

Experimental Hematology

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“…Normal control samples were derived from malignant lymphoma patients without bone marrow involvement as described earlier. 24,25 Our analysis of WT1, GATA-1 and GATA-2 was approved by IRB (approved number 99) of the Nagoya University School of Medicine. Some leukemia cell lines were provided by Dr Hirokazu Nagai (National Hospital Organization, Nagoya Medical Center, Nagoya, Japan).…”

Section: Clinical Samples and Cell Linesmentioning

confidence: 99%

“…24,25 WT1 mRNA was measured using the Light Cycler system (Roche Diagnostics, Mannheim, Germany). Each primer set and PCR conditions have been described before.…”

Section: Quantitative Reverse Transcriptase-pcrmentioning

confidence: 99%

“…Each primer set and PCR conditions have been described before. 24,25 GATA-1, GATA-2 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNAs were measured with a Power SYBR Green master mix using the ABI PRISM 7000 sequence detection systems (Applied Biosystems Japan Ltd, Tokyo, Japan). After producing first-strand cDNA, quantitative PCR was carried out using the following primer sets: GATA-1, forward 5 0 -CCCAAGAAGCGCCT GATTGT-3 0 ; reverse 5 0 -GTGTAGCTTGTAGTAGAGGCCGC-3 0 .…”

Section: Quantitative Reverse Transcriptase-pcrmentioning

confidence: 99%

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GATA-1 and GATA-2 binding to 3′ enhancer of WT1 gene is essential for its transcription in acute leukemia and solid tumor cell lines

et al. 2009

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Although oncogenic functions and the clinical significance of Wilms tumor 1 (WT1) have been extensively studied in acute leukemia, the regulatory mechanism of its transcription still remains to be determined. We found a significant correlation among the amounts of WT1, GATA-1 and GATA-2 mRNAs from leukemia and solid tumor cell lines. Overexpression and small interfering RNA (siRNA) transfection experiments of GATA-1 and GATA-2 showed that these GATA transcription factors could induce WT1 expression. Promoter analysis showed that the 5 0 promoter did not explain the different WT1 mRNA levels between cell lines. The 3 0 enhancer, especially the distal sites out of six putative GATA binding sites located within the region, but not the intron 3 enhancer, were essential for the WT1 mRNA level. Electrophoretic mobility shift assay (EMSA) showed both GATA-1 and GATA-2 bound to these GATA sites. Besides acute leukemia cell lines, solid tumor cell lines including, TYK-nu-cPr also showed a high level of WT1 mRNA. We showed that GATA-2 expression is a determinant of WT1 mRNA expression in both TYK-nu-cPr cells and HL60 cells without GATA-1 expression. Taken together, these results suggest that GATA-1 and/or GATA-2 binding to a GATA site of the 3 0 enhancer of WT1 played an important role in WT1 gene expression.

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“…However, elevated expression of sphingosine kinase has been reported in multiple types of cancer. The mRNA levels of sphingosine kinase were significantly increased in breast, colon, lung, ovary, uterus, and kidney cancer patients, as well as in acute leukemia patients [2,7,23].…”

Section: Discussionmentioning

confidence: 99%

Differential Cytotoxic Effects of Jaspine B in Various Cancer Cells

Lee¹,

Choi²,

Kwon³

et al. 2016

Journal of Life Science

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Jaspine B is an anhydrophytosphingosine that is isolated from a marine sponge. Because of its structural similarity to sphingosine, it shows anti-cancer effects in human carcinomas. Therefore, this study aims to investigate its anti-proliferative effect on various cancer cells and to correlate its association with the intracellular accumulation of Jaspine B in relevant cancer cells. The anti-proliferative effect of Jaspine B in various cancer cells was determined by a cell viability test, and the intracellular concentration of Jaspine B in relevant cancer cells was determined using mass spectrometry coupled with liquid chromatography. The correlation coefficient and p value between the cytotoxicity and the cell accumulation of Jaspine B were determined using SPSS 16.1. The cytotoxicity of Jaspine B varied depending on the type of cancer cell when compared the EC50 values of Jaspine B. Breast and melanoma cancer cells were susceptible to Jaspine B, whereas renal carcinoma cells were resistant. The intracellular concentrations of Jaspine B had a reciprocal correlation with the EC50 values in the same cells (r = 0.838). The results suggested that the anti-proliferative effect of Jaspine B was associated with the cellular accumulation of this compound. However, Jaspine B was not a substrate for P-glycoprotein and breast cancer resistance protein, as major efflux pumps caused multidrug resistance. The maintenance of a high intracellular concentration is crucial for the cytotoxic effect of Jaspine B; however, efflux pumps may not be a controlling factor for Jaspine B-related resistance in cancer cells.

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Quantitative RT-PCR analysis of sphingolipid metabolic enzymes in acute leukemia and myelodysplastic syndromes

Cited by 58 publications

References 6 publications

Sphingosine kinase 1 overexpression is regulated by signaling through PI3K, AKT2, and mTOR in imatinib-resistant chronic myeloid leukemia cells

Sphingosine kinase 1 overexpression is regulated by signaling through PI3K, AKT2, and mTOR in imatinib-resistant chronic myeloid leukemia cells

GATA-1 and GATA-2 binding to 3′ enhancer of WT1 gene is essential for its transcription in acute leukemia and solid tumor cell lines

Differential Cytotoxic Effects of Jaspine B in Various Cancer Cells

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