[Show abstract][Hide abstract] ABSTRACT: Epigenetic silence in cancer frequently altered signal-transduction pathways during the early stages of tumor development. Recent progress in the field of cancer epigenetics has led to new opportunities for diagnosis and treatment of cancer. We previously demonstrated that novel identified nuclear factor MARVELD1 was widely expressed in human tissues, but down-regulated by promoter methylation in multiple cancers. This study was carried out to determine the biological and clinical significance of MARVELD1 gene silencing in lung cancer. Here, we found the reduced MARVELD1 expression significantly correlated with diagnostic histopathology and malignant degree of lung cancers. DNA hypermethylation and histone deacetylation synergistically inactivated MARVELD1 gene in lung cancer cells. Moreover, MARVELD1 modulated the efficiency of nonsense-mediated mRNA decay (NMD) through interaction with NMD core factor SMG1. The decreased MARVELD1 level in lung cancer reduces NMD efficiency through diminishing the association between NMD complex component UPF1/SMG1 and premature termination codons containing mRNA (PTC-mRNA). The results suggested that MARVELD1 silencing is an appealing diagnostic biomarker for lung cancer and epigenetic silencing of MARVELD1 gene links with the regulatory mechanism of NMD pathway in lung cancer, which may be required for tumorigenesis.
[Show abstract][Hide abstract] ABSTRACT: Apoptosis-inducing factor (AIF) plays a crucial role in caspase-independent programmed cell death by triggering chromatin condensation and DNA fragmentation. Therefore, it might be involved in cell homeostasis and tumor development. In this study, we report significant AIF downregulation in the majority of renal cell carcinomas (RCC). In a group of RCC specimens, 84% (43 out of 51) had AIF downregulation by immunohistochemistry stain. Additional 10 kidney tumors, including an oxyphilic adenoma, also had significant AIF downregulation by Northern blot analysis. The mechanisms of the AIF downregulation included both AIF deletion and its promoter methylation. Forced expression of AIF in RCC cell lines induced massive apoptosis. Further analysis revealed that AIF interacted with STK3, a known regulator of apoptosis, and enhanced its phosphorylation at Thr180. These results suggest that AIF downregulation is a common event in kidney tumor development. AIF loss may lead to decreased STK3 activity, defective apoptosis and malignant transformation.
PLoS ONE 07/2014; 9(7):e100824. DOI:10.1371/journal.pone.0100824 · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Cell adhesion on an extracellular matrix (ECM) participates in cell motility, invasion, cell signal transduction and gene expression. Many nuclear proteins regulate cell-ECM adhesion through managing the transcription of cell adhesion-related genes. Here, we identified MARVEL [MAL (The myelin and lymphocyte protein) and related proteins for vesicle trafficking and membrane link] domain containing 1 (MARVELD1) that could suppress cell spreading and complicate actin organization. Over-expression of MARVELD1 in NIH3T3 cells decreased the expression level of integrin β1 and vinculin, and further led to dephosphorylation of focal adhesion kinase (FAK) at Tyr 397. We also found that MARVELD1 partially colocalized with serine/arginine-rich splicing factor 2 (SC35) and interacted with nuclear cap binding protein subunit 2 (CBP20). Finally, we demonstrated that pre-mRNA processing of integrin β1 was affected by MARVELD1. Taken together, our studies demonstrate that MARVELD1 plays a role in pre-mRNA processing of integrin β1, and thereby regulates cell adhesion and cell motility. These studies provide a novel regulatory mechanism of cell-ECM adhesion by nuclear protein in cells.
The international journal of biochemistry & cell biology 09/2013; 45(11). DOI:10.1016/j.biocel.2013.09.006 · 4.24 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The complex space environments can influence cell structure and function. The research results on space biology have shown that the major mutagenic factors in space are microgravity and ionizing radiation. In addition, possible synergistic effects of radiation and microgravity on human cells are not well understood. In this study, human immortal lymphoblastoid cells were established from human peripheral blood lymphocytes and the cells were treated with low dose (0.1, 0.15 and 0.2 Gy) cumulative 60Co γ-irradiation and simulated weightlessness [obtained by culturing cells in the Rotating Cell Culture System (RCCS)]. The commonly used indexes of cell damage such as micronucleus rate, cell cycle and mitotic index were studied. Previous work has proved that Gadd45 (growth arrest and DNA-damage-inducible protein 45) gene increases with a dose-effect relationship, and will possibly be a new biological dosimeter to show irradiation damage. So Gadd45 expression is also detected in this study. The micronucleus rate and the expression of Gadd45α gene increased with irradiation dose and were much higher after incubation in the rotating bioreactor than that in the static irradiation group, while the cell proliferation after incubation in the rotating bioreactor decreased at the same time. These results indicate synergetic effects of simulated weightlessness and low dose irradiation in human cells. The cell damage inflicted by γ-irradiation increased under simulated weightlessness. Our results suggest that during medium- and long-term flight, the human body can be damaged by cumulative low dose radiation, and the damage will even be increased by microgravity in space.
[Show abstract][Hide abstract] ABSTRACT: This study investigated intracellular oxidative stress and its underlying mechanisms in a rotary cell culture system used to achieve a simulated microgravity (SMG) environment. Experiments were conducted with human breast cancer cell lines MCF-7 (an estrogen receptor (ER) α positive cell line) and MDA-MB-231 (an ERα negative cell line) encapsulated in alginate/collagen carriers. After 48 h, SMG led to oxidative stress and DNA damage in the MDA-MB-231 cells but a significant increase in mitochondrial activity and minimal DNA damage in the MCF-7 cells. The activity of superoxide dismutase (SOD) significantly increased in the MCF-7 cells and decreased in MDA-MB-231 cells in the SMG environment compared with a standard gravity control. Moreover, SMG promoted expression of ERα and protein kinase C (PKC) epsilon in MCF-7 cells treated with PKC inhibitor Gö6983. Overall, exposure to SMG increased mitochondrial activity in ERα positive cells but induced cellular oxidative damage in ERα negative cells. Thus, ERα may play an important role in protecting cells from oxidative stress damage under simulated microgravity.
Advances in Space Research 05/2012; 49(10):1432–1440. DOI:10.1016/j.asr.2012.02.020 · 1.35 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In the present study we identified a novel gene, Homo Sapiens Chromosome 1 ORF109 (c1orf109, GenBank ID: NM_017850.1), which encodes a substrate of CK2. We analyzed the regulation mode of the gene, the expression pattern and subcellular localization of the predicted protein in the cell, and its role involving in cell proliferation and cell cycle control.
Dual-luciferase reporter assay, chromatin immunoprecipitation and EMSA were used to analysis the basal transcriptional requirements of the predicted promoter regions. C1ORF109 expression was assessed by western blot analysis. The subcellular localization of C1ORF109 was detected by immunofluorescence and immune colloidal gold technique. Cell proliferation was evaluated using MTT assay and colony-forming assay.
We found that two cis-acting elements within the crucial region of the c1orf109 promoter, one TATA box and one CAAT box, are required for maximal transcription of the c1orf109 gene. The 5' flanking region of the c1orf109 gene could bind specific transcription factors and Sp1 may be one of them. Employing western blot analysis, we detected upregulated expression of c1orf109 in multiple cancer cell lines. The protein C1ORF109 was mainly located in the nucleus and cytoplasm. Moreover, we also found that C1ORF109 was a phosphoprotein in vivo and could be phosphorylated by the protein kinase CK2 in vitro. Exogenous expression of C1ORF109 in breast cancer Hs578T cells induced an increase in colony number and cell proliferation. A concomitant rise in levels of PCNA (proliferating cell nuclear antigen) and cyclinD1 expression was observed. Meanwhile, knockdown of c1orf109 by siRNA in breast cancer MDA-MB-231 cells confirmed the role of c1orf109 in proliferation.
Taken together, our findings suggest that C1ORF109 may be the downstream target of protein kinase CK2 and involved in the regulation of cancer cell proliferation.
[Show abstract][Hide abstract] ABSTRACT: We have previously found that expression of MARVELD1 was remarkably downregulated in multiple tumor tissues, but unclear in hepatocellular carcinoma (HCC) and its function has not been explored yet. In the present study, to uncover the underlying mechanism of MARVELD1 in the pathogenesis and development of HCC, we investigated the expression pattern of MARVELD1 and its effect on tumor proliferation in HCC. The results indicated the frequent downregulation of MARVELD1 in clinic samples and cell lines of HCC resulted from promoter methylation, as well as genetic deletion. Furthermore, treatment of MARVELD1 unexpressing Hep3B2.1-7 and PLC/PRF/5 cells with the demethylating agent 5-aza-2' deoxycytidine restored its expression. Overexpression of MARVELD1 suppressed the proliferation of HCC cells in vitro and in vivo, whereas downregulation of endogenous MARVELD1 by shRNAs significantly enhanced these characters. MARVELD1 overexpression could enhance chemosensitivity of HCC cells to epirubicin and 10-hydroxycamptothecin. Corresponding to these results, the expression of p-ERK1/2 and cyclin D1 were decreased, whereas p16 and p53 were increased in MARVELD1-transfected cells. We also demonstrated that knockdown of MARVELD1 resulted in upregulation of p-ERK1/2 and cyclin D1, and downregulation of p16 and p53. Moreover, the effect of the decreased cell growth rate was significantly reversed when MARVELD1-overexpressing cells were trasfected with p53 or p16 siRNA. Our findings suggest that MARVELD1 is a tumor suppressor by negatively regulating proliferation, tumor growth and chemosensitivity of HCC cells via increasing p53 and p16 in vitro and in vivo. MARVELD1 may be a potential target for HCC therapy.
Cancer Science 02/2012; 103(4):716-22. DOI:10.1111/j.1349-7006.2012.02220.x · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Rotary cell culture systems (RCCS) have been shown to be promising for promoting three-dimensional (3D) cell growth and assembly of cells into functional tissues. In this study, 3D tissue-like spheroids of MCF-7 cells were constructed by encapsulating the cells in the collagen-alginate hydrogel, and then cultured in a RCCS to investigate the proliferation of MCF-7 cells. The results from the MTT assay showed that the proliferation rate of MCF-7 cells cultured in the RCCS was higher than that of the static culture control group, and the results from the flow cytometry revealed that the cells in S and G2/M phase were significantly increased compared to the control group. The expression of cell proliferation antigen PCNA and cyclin D1 was also examined with the results further supporting the enhanced proliferation of MCF-7 cells by the RCCS. The results from indirect immunofluorescence revealed that the rotary culture altered neither the cytoskeleton distribution nor the assembly of mitotic spindle. By examination, it was also shown that the rotary culture induced the ERK1/2-MAPK pathway. Taken together, this study demonstrated that the rotary culture could promote the proliferation of MCF-7 cells by inducing the ERK1/2 pathway.
[Show abstract][Hide abstract] ABSTRACT: MARVEL domain-containing 1 (MARVELD1) is a newly identified nuclear protein; however its function has not been clear until now. Here, we report that mouse MARVELD1 (mMARVELD1), which is highly conserved between mice and humans, exhibits cell cycle-dependent cellular localization. In NIH3T3 cells, MARVELD1 was observed in the nucleus and at the perinuclear region during interphase, but was localized at the mitotic spindle and midbody at metaphase, and a significant fraction of mMARVELD1 translocated to the plasma membrane during anaphase. In addition, treatment of cells with colchicine, a microtubule-depolymerizing agent, resulted in translocation of mMARVELD1 to the plasma membrane, and association of mMARVELD1 and α-tubulin was confirmed by co-immunoprecipitation. Finally, overexpression of mMARVELD1 resulted in a remarkable inhibition of cell proliferation, G1-phase arrest, and reduced cell migration. These findings indicate that mMARVELD1 is a microtubule-associated protein that plays an important role in cell cycle progression and migration.
[Show abstract][Hide abstract] ABSTRACT: The main induce factors of the space environment are sustaining microgravity and ionizing radiation which can influence cell structure and function. In this study, for investigating the damage to human proliferation cells under microgravity effects, human osteosarcoma cell lines (MG-63 and U-2 OS) and peripheral blood lymphocytes were analyzed after ground-based simulated microgravity effect with clinostat. The present results showed that under the simu-lated microgravity effect for 72 hours the cytoskeleton both in human osteosarcoma cell lines and peripheral blood lymphocytes were in disorder and the spindle structure changed. The multiple polar spindle rates in human osteosarcoma cell lines MG-63 and U-2 OS increased in a linear relationship with simulated microgravity hours (0 to 96 hours). The mitotic index increased in human osteosarcoma cell lines MG-63 and U-2 OS, while decreased in human pe-ripheral blood lymphocytes. And the cell cycle of MG-63 arrest at G2/M phase. The mode number of chromosomes also varied in MG-63, while the chromosome number of human periph-eral blood lymphocytes didn't show significant difference. The Real-time quantitative reverse transcription PCR (qRT-PCR) and Western blot showed that the spindle assembly checkpoint protein MAD2 and BUB1 in MG-63 and U-2 OS changed with the spindle structure change and chromosome number change.
[Show abstract][Hide abstract] ABSTRACT: Grb10 gene expression was analyzed in fetus and four organs during gestation process using the real-time RT-PCR and in situ hybridization. The real-time RT-PCR analysis result showed that the expression of Grb10 in whole embryos is throughout E8.5 to E19.5. The Grb10 gene expression level was gradually increased from E8.5 to E13.5, and then gradually reduced. The tissues specific expression levels were decreased in brain, heart, and lung from E12.5 to E19.5, but a peak expression was observed in liver at E18.5. Grb10 gene expression was also investigated at E13.5, E15.5, E16.5 and E18.5 sections by using in situ hybridization analysis. Strong signals of Grb10 were detected in brain, spine, kidney, and muscle tissues. These results suggest that Grb10 may play an important role during the development process in mouse.
[Show abstract][Hide abstract] ABSTRACT: MARVELD1 (MARVEL domain-containing 1) is a member of MARVEL domain-containing proteins and located on human chromosome 10q24.2. MARVELD1 has no significant similarity with other members of MARVEL domain family at amino acid level. Gene expression arrays demonstrated that MARVELD1 is widely expressed in normal human tissues and is down-regulated in primary multiple tumors derived from ovary, vulva, uterus, cervix, breast, testis, kidney, bladder and liver. The down-regulation of MARVELD1 was further identified by real-time PCR and immunohistochemical staining in primary breast cancer. In addition, we identified the reduced expression of MARVELD1 is owing to DNA methylation and could be reversed by pharmacologic demethylation. Finally, our results showed that MARVELD1 protein is located in nucleus instead of cell membrane.
Cancer letters 05/2009; 282(1):77-86. DOI:10.1016/j.canlet.2009.03.008 · 5.62 Impact Factor