SPAG6 silencing inhibits the growth of the malignant myeloid cell lines SKM-1 and K562 via activating p53 and caspase activation-dependent apoptosis

Yang, Bihui; Wang, Li; Luo, Xiao‐Hua; Chen, Liping; Zhou, Yang; Liu, Lin

doi:10.3892/ijo.2014.2768

Cited by 23 publications

(35 citation statements)

References 33 publications

(29 reference statements)

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“…However, in this study, SKM-1 cells were subcutaneously injected and solid tumors were formed in NOD/SCID mice. The same phenomenon was observed in another study (28), in which both SKM-1 cells and K562 cells were transplanted subcutaneously and solid tumors formed in immunodeficient mice. This discrepancy may attribute to the fact that the microenvironments provided by these mice are not identical to those for humans, although they are suitable for MDS cell-derived tumor growth.…”

Section: Discussionsupporting

confidence: 81%

“…Protein lysate was extracted from cells using RIPA lysis buffer supplemented with 1 µM PMSF (Beyotime), and protein concentration was measured with BCA Protein Assay kit (Beyotime). Western blotting was performed as previously described (28). The following primary antibodies were used in this study: rabbit anti-human cleaved-caspase-3, cleaved-caspase-8, cleaved-caspase-9, Bcl-w and Bax (immunoway, Newark, DE, USA); rabbit polyclonal antibody against human CDC40 (Abcam, Cambridge, UK); horseradish peroxidase-conjugated goat anti-rabbit IgG (Beyotime) was used as second antibody in this study.…”

Section: Quantitative Reverse Transcriptase-polymerase Chain Reactionmentioning

confidence: 99%

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miR-378 inhibits cell growth and enhances apoptosis in human myelodysplastic syndromes

Kuang

Wei

Zhang

et al. 2016

International Journal of Oncology

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miR-378 has been proven to inhibit cell growth, migration and invasion in different types of cancers. In this study, we found that miR-378 was commonly downregulated in the bone marrow cells obtained from myelodysplastic syndrome (MDS) patients. We further investigated the role of miR-378 in the proliferation and apoptosis of SKM-1 cells, an acute myeloid leukemia cell line established in the leukemic phase during the progression of MDS to AML (MDS/AML). Results indicated that overexpression of miR-378 in SKM-1 cells interfered with proliferation via inducing apoptosis and G0/G1-phase cell cycle arrest, and suppressive effect of miR-378 on MDS/AML cells may be mediated partly through Bcl-w and CDC40. Moreover, apoptosis induced by miR-378 correlated with increased expression of Bax and activation of caspase-3, -8 and -9. Taken together, our data support a critical role for miR-378 in the pathogenesis of MDS and provide a novel therapeutic target in this complex disease.

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Section: Discussionsupporting

confidence: 81%

Section: Quantitative Reverse Transcriptase-polymerase Chain Reactionmentioning

confidence: 99%

miR-378 inhibits cell growth and enhances apoptosis in human myelodysplastic syndromes

Kuang

Wei

Zhang

et al. 2016

International Journal of Oncology

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“…A preliminary study indicated that a high expression of SPAG6 is connected with apoptosis resistance in SKM-1 and K562 (13). In this study, when SKM-1 cells were treated with different concentrations of rTRAIL, the apoptosis rates showed an increased tendency.…”

Section: Discussionmentioning

confidence: 60%

“…Furthermore, SPAG6 is highly expressed at the break point of t(10;11)(p12;q14) in patients with CALM/AF10-positive leukemias (12). In our previous study, when the expression of SPAG6 was silenced by SPAG6-shRNA lentivirus, the growth of SKM-1 and K652 was markedly inhibited, and apoptosis was increased (13). The above evidence indicates that SPAG6 may promote cellular growth by preventing apoptosis.…”

Section: Introductionmentioning

confidence: 83%

SPAG6 regulates cell apoptosis through the TRAIL signal pathway in myelodysplastic syndromes

Yang

Wang

et al. 2017

Oncology Reports

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Myelodysplastic syndromes (MDSs) are a group of malignant clone hematopoietic stem-cell diseases, and the evolution and progression of MDS depend on the abnormal apoptosis of bone marrow cells. Our previous studies have indicated that sperm-associated antigen 6 (SPAG6), located in the uniparental disomy regions of myeloid cells, is overexpressed in patients with MDS as compared to controls, and SPAG6 can inhibit apoptosis of SKM-1. However, the concrete mechanism is still unclear. In the present study, it was found that the TNF-related apoptosis-inducing ligand (TRAIL)signal pathway was activated when the expression of SPAG6 was inhibited by SPAG6-shRNA lentivirus in SKM-1 cells. Additionally, the results of flow cytometry, Cell Counting Kit-8 assay and western blot analysis implied that the TRAIL signal pathway could be inhibited by a high expression of SPAG6. However, SPAG6 cannot influence the expression of TRAIL death receptors, except for FADD. Additionally the interaction between FADD and TRAIL death receptors also increased in SKM-1 cells infected with SPAG6-shRNA lentivirus. Thus, our study demonstrates that SPAG6 may regulate apoptosis in SKM-1 through the TRAIL signal pathway, indicating that SPAG6 could be a potential therapeutic target.

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“…Based on all these observations, we selected the genes SPAG6 (Sperm Associated Antigen 6) and L1TD1 (LINE-1 Type Transposase Domain Containing 1) for detailed investigation. SPAG6 is located in the chromosomal region 10p12.2 and is thought to be a cancer-testis antigen (CTA) [18]. CTAs represent a large family of cancer-associated antigens which are expressed in immunoprivileged tissues such as testis but were also detected in tumor tissues of various origins including lung cancer [19].…”

Section: Introductionmentioning

confidence: 99%

SPAG6 and L1TD1 are transcriptionally regulated by DNA methylation in non-small cell lung cancers

et al. 2017

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BackgroundDNA methylation regulates together with other epigenetic mechanisms the transcriptional activity of genes and is involved in the pathogenesis of malignant diseases including lung cancer. In non-small cell lung cancer (NSCLC) various tumor suppressor genes are already known to be tumor-specifically methylated. However, from the vast majority of a large number of genes which were identified to be tumor-specifically methylated, tumor-specific methylation was unknown so far. Thus, the major aim of this study was to investigate in detail the mechanism(s) responsible for transcriptional regulation of the genes SPAG6 and L1TD1 in NSCLCs.MethodsWe analysed publically available RNA-sequencing data and performed gene expression analyses by RT-PCR. DNA methylation analyses were done by methylation-sensitive high-resolution melt analyses and bisulfite genomic sequencing. We additionally investigated protein expression using immunohistochemistry. Cell culture experiments included tumor cell growth, proliferation, viability as well as colony formation assays. Moreover, we performed xenograft experiments using immunodeficient mice.ResultsWe observed frequent downregulation of SPAG6 and L1TD1 mRNA expression in primary tumor (TU) samples compared to corresponding non-malignant lung tissue (NL) samples of NSCLC patients. We furthermore observed re-expression of both genes after treatment with epigenetically active drugs in most NSCLC cell lines with downregulated SPAG6 and L1TD1 mRNA expression. Frequent tumor-specific DNA methylation of SPAG6 and L1TD1 was detected when we analysed TU and corresponding NL samples of NSCLC patients. ROC curve analyses demonstrated that methylation of both genes is able to distinguish between TU and NL samples of these patients. Immunohistochemistry revealed a close association between SPAG6/L1TD1 methylation and downregulated protein expression of these genes. Moreover, by performing functional assays we observed reduced cell growth, proliferation and viability of pCMV6-L1TD1 transfected NSCLC cells. In addition, reduced volumes of tumors derived from pCMV6-L1TD1 compared to pCMV6-ENTRY transfected NCI-H1975 cells were seen in a xenograft tumor model.ConclusionsOverall, our results demonstrate that SPAG6 and L1TD1 are tumor-specifically methylated in NSCLCs and that DNA methylation is involved in the transcriptional regulation of these genes. Moreover, in vitro as well as in vivo experiments revealed tumor-cell growth suppressing properties of L1TD1 in NSCLC cells.Electronic supplementary materialThe online version of this article (doi:10.1186/s12943-016-0568-5) contains supplementary material, which is available to authorized users.

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SPAG6 silencing inhibits the growth of the malignant myeloid cell lines SKM-1 and K562 via activating p53 and caspase activation-dependent apoptosis

Cited by 23 publications

References 33 publications

miR-378 inhibits cell growth and enhances apoptosis in human myelodysplastic syndromes

miR-378 inhibits cell growth and enhances apoptosis in human myelodysplastic syndromes

SPAG6 regulates cell apoptosis through the TRAIL signal pathway in myelodysplastic syndromes

SPAG6 and L1TD1 are transcriptionally regulated by DNA methylation in non-small cell lung cancers

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