| 1. |
- Attarha, S., et al.
(författare)
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PKN I modulates TGF beta and EGF signaling in HEC-I-A endometrial cancer cell line
- 2014
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Ingår i: Oncotargets and Therapy. - : Informa UK Limited. - 1178-6930. ; 7, s. 1397-1408
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Tidskriftsartikel (refereegranskat)abstract
- Background: The response of cells to TGF beta and EGF is mediated by a network of various intracellular regulators. The signaling crosstalk between different regulators is of key importance for tumorigenesis. The crosstalk may explain the modulation of cellular responses to the same regulator by another signaling molecule. As PKN1-a serine/threonine kinase implicated in tumorigenesis was identified as potential crosstalk node for TGF beta and EGF signaling, the cellular functions that may be affected by PKN1 in a crosstalk of TGF beta and EGF were explored. Methods: To investigate the contribution of PKN1 to TGF beta and EGF signaling, transiently PKN1-transfected HEC-1-A endometrial cancer cells were generated and subjected to treatment with TGF beta, EGF, and their combination. Proliferation, apoptosis, invasion, wound healing, and migration assays were performed. The impact of PKN1 on the expression and phosphorylation of intracellular proteins was monitored by immunoblotting. Results: It was demonstrated that PKN1 modulated the responses of HEC-A-1 endometrial cancer cells to TGF beta and EGF PKN1 had an inhibitory effect on the stimulation of cell migration, and PKN1 kinase activity was required for the inhibitory effect of TGF beta and EGF on cell proliferation and invasiveness. It was observed that phosphorylation of Smad2, FAK, and Erk1/2 correlated with responses of the cells to TGF beta and EGE Conclusion: PKN1 modulates TGF beta- and EGF-dependent regulation of cell proliferation, migration, and invasiveness, and therefore is a component of the network signaling downstream of IGO and EGE
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| 2. |
- Chavali, Pavithra Lakshminarasimhan, et al.
(författare)
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Nuclear orphan receptor TLX induces Oct-3/4 for the survival and maintenance of adult hippocampal progenitors upon hypoxia.
- 2011
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Ingår i: The Journal of biological chemistry. - 1083-351X. ; 286:11, s. 9393-9404
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Tidskriftsartikel (refereegranskat)abstract
- Hypoxia promotes neural stem cell proliferation, the mechanism of which is poorly understood. Here, we have identified the nuclear orphan receptor TLX as a mediator for proliferation and pluripotency of neural progenitors, upon hypoxia. We find an enhanced early protein expression of TLX under hypoxia potentiating sustained proliferation of neural progenitors. Moreover, TLX induction upon hypoxia in differentiating conditions leads to increased proliferation and stem cell like phenotype along with coexpression with neural stem cell markers. Following hypoxia, TLX is recruited to Oct-3/4 proximal promoter, augmenting the gene transcription and promoting progenitor proliferation and pluripotency. Knock-down of Oct-3/4 significantly reduced TLX mediated proliferation, highlighting their interdependence in regulating progenitor pool. Additionally, TLX synergizes with bFGF to sustain cell viability upon hypoxia, since the knock-down of TLX along with the withdrawal of growth factor results in cell death. This can be attributed to the activation of Akt signaling pathway by TLX, the depletion of which results in reduced proliferation of progenitor cells. Cumulatively, the data presented here demonstrate a new role for TLX in neural stem cell proliferation and pluripotency upon hypoxia.
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| 3. |
- Chavali, Pavithra Lakshminarasimhan, et al.
(författare)
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TLX activates MMP-2, promotes self-renewal of tumor spheres in neuroblastoma and correlates with poor patient survival.
- 2014
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Ingår i: Cell Death & Disease. - : Springer Science and Business Media LLC. - 2041-4889. ; 449:5
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Tidskriftsartikel (refereegranskat)abstract
- Nuclear orphan receptor TLX (Drosophila tailless homolog) is essential for the maintenance of neural stem/progenitor cell self-renewal, but its role in neuroblastoma (NB) is not well understood. Here, we show that TLX is essential for the formation of tumor spheres in three different NB cell lines, when grown in neural stem cell media. We demonstrate that the knock down of TLX in IMR-32 cells diminishes its tumor sphere-forming capacity. In tumor spheres, TLX is coexpressed with the neural progenitor markers Nestin, CD133 and Oct-4. In addition, TLX is coexpressed with the migratory neural progenitor markers CD15 and matrix metalloproteinase-2 (MMP-2) in xenografts of primary NB cells from patients. Subsequently, we show the effect of TLX on the proliferative, invasive and migratory properties of IMR-32 cells. We attribute this to the recruitment of TLX to both MMP-2 and Oct-4 gene promoters, which resulted in the respective gene activation. In support of our findings, we found that TLX expression was high in NB patient tissues when compared with normal peripheral nervous system tissues. Further, the Kaplan-Meier estimator indicated a negative correlation between TLX expression and survival in 88 NB patients. Therefore, our results point at TLX being a crucial player in progression of NB, by promoting self-renewal of NB tumor-initiating cells and altering their migratory and invasive properties.
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| 4. |
- Kolakowska, J., et al.
(författare)
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Progress and challenges in the proteomics of domestic pig in research on the female reproductive system
- 2016
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Ingår i: Journal of Elementology. - : Polish Society for Magnesium Research. - 1644-2296. ; 21:4, s. 1055-1069
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Tidskriftsartikel (refereegranskat)abstract
- The proteomics of pigs is developing dynamically, attracting much attention among representatives of medical and health sciences, veterinary medicine, agricultural and natural sciences. The pig has been widely studied in genetics and genomics. However, proteomic applications are still awaiting more extensive implementation, both in the use of pigs as animal models in biomedicine sciences, and in the exploration of physiological tracts important for pig production. Underdeveloped databases for identification and analysis of porcine proteins as well as the scarcity of detailed information on similarities and differences between humans and pigs at the molecular level are hampering the pig proteomics. However, the use of pigs in proteomic studies in both physiological and biomedical sciences is prevalent comparing to other farm animals. The focus of the reported pig model proteomics studies is on exploring physiology and diseases, and on improving pig breading and productivity. This species has been used as a model in proteomics studies involved in ocular, brain, nutritional and reproduction research, etc. In the present paper we discuss technologies and bioinformatic tools used in studies of a proteome to verify the peptide- and protein-based content and we summarize the current status of proteomic studies of pigs. We focus on studies of the female reproductive system because the examination and understanding of the biology of oocytes, the oviduct and the uterus could facilitate the identification of mechanisms involved in the prenatal development, and it may help to develop new treatment for infertility of farm animals. © 2016, Polish Society Magnesium Research. All rights reserved.
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| 5. |
- Kolakowska, J., et al.
(författare)
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Proteomic analysis of the endometrium during early pregnancy in the domestic pig
- 2017
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Ingår i: Reproduction Fertility and Development. - : CSIRO Publishing. - 1031-3613. ; 29:11, s. 2255-2268
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Tidskriftsartikel (refereegranskat)abstract
- Reproductive processes in domestic pigs have been studied extensively. Pigs are one of the main sources of meat for human consumption and are an established model for investigations into mammalian, including human, reproductive physiology. Studies of the uterus during early pregnancy will lead to a better understanding of mechanisms governing pregnancy. Proteomics provides the possibility to explore endometrial functions in an unbiased way. The aim of the study was to compare endometrium harvested from Days 12-13 and 15-16 of pregnancy with the corresponding days of the oestrous cycle. We identified endometrial proteins that are unique to the early stages of pregnancy (Days 12-13 and 15-16). Twenty-one proteins were identified that were uniquely expressed on the selected days of pregnancy or the oestrous cycle. Out of 21 identified proteins, 14 referred to the pregnancy periods. Systemic analysis of the identified proteins revealed cell adhesion and cytoskeletal organisation as two of the major functions, both of which are important for the establishment and maintenance of pregnancy. Thrombospondin 1 expression was validated using western blotting analysis and the results suggest its involvement in the adhesiveness of the embryo during the peri-implantation period in pigs.
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| 6. |
- Saini, Ravi Kanth Rao
(författare)
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Neural stam cells, TLX, and Neuroblastoma
- 2014
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Neuroblastoma (NB) is one of the most commonly diagnosed extracranial tumors in children below five years of age. Moreover, NB accounts for almost 12-15% of all childhood cancer related fatalities. To date, the origin of NB has been linked to the neural crest derived sympathoadrenal progenitor cells. Progression and pathogenicity of the disease has been largely correlated with the ages of children and presence of undifferentiated neuroblasts and amplification of the MYCN oncogene. However from the clinical evidence gathered so far, MYCN amplification is related to poor prognosis but it occurs only in 20% of NB cases. Henceforth, there is a need for better diagnostic and predictive markers to stratify NB patients. NB cells express a number of neural stem cell and progenitor markers such as Oct3/4, Sox2, CD133, ABCG2, and Nestin. Since self-renewal and differentiation of neural stem cells are predominantly regulated by a number of stem cell fate determinants such as Notch, Wnt, Hedgehog, PTEN, and TLX, it is speculated that deregulation of these genes may be responsible for the pathogenicity of the disease. TLX is an orphan nuclear receptor, which is predominantly expressed in the embryonic and adult forebrain, and is considered to be a crucial regulator of neurogenesis, because of its roles in neural stem cell self-renewal and maintenance. We have identified that upon hypoxia, TLX stimulates neural stem cell renewal by promoting Oct3/4 transcription in adult hippocampal progenitors. TLX is expressed at high levels in NB cell lines. In sphere forming cells generated from these cell lines, TLX was enriched and co- expressed along with the neural progenitor markers Nestin, Oct3/4, CD133, and HIF- 2α. TLX was also co-expressed with the neural progenitor markers CD15 and MMP-2 in xenografts of primary NB-tumor initiating cells (TIC) derived from patients. Thus, TLX may be involved in the tumorigenesis of NB by promoting dedifferentiation of tumor cells. NB develops through processes which may be defined as cellular “dedifferentiation”. The ability of tumors to form spheroids is one of the manifestations of dedifferentiation and transformation. To study the mechanisms of dedifferentiation, neuroblastoma cell lines will help us to identify new diagnostic markers. We generated spheroids of the NB cell line SK-N-BE2 and performed proteomics analysis to evaluate the differential expression pattern as compared with the wild-type cells. We identified 239 proteins which were affected by the dedifferentiation process. These proteins represented several regulatory processes, such as transcription, cell cycle regulation, apoptosis, cell adhesion, metabolism, intracellular transport, stress response, and angiogenesis. An extensive analysis using Cytoscape identified “DISC-1” and “DNA-PKcs”, both of which have been previously linked to dedifferentiation and cancer. The results contribute to better understanding of mechanisms involved in Neuroblastoma pathogenesis, along with identifying possible biomarkers for the disease that may be translated to the clinic.
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| 7. |
- Saini, Ravi Kanth Rao, et al.
(författare)
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Proteomics of dedifferentiation of SK-N-BE2 neuroblastoma cells
- 2014
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Ingår i: Biochemical and Biophysical Research Communications. - : Elsevier BV. - 0006-291X .- 1090-2104. ; 454:1, s. 202-209
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Tidskriftsartikel (refereegranskat)abstract
- Neuroblastoma develops through processes which include cellular dedifferentiation. Ability of tumors to form spheroids is one of the manifestations of dedifferentiation and carcinogenic transformation. To study mechanisms of dedifferentiation of neuroblastoma cells, we generated spheroids and performed a proteomics study to compare the spheroids with parental SK-N-BE2 cells. We observed that dedifferentiation induced extensive changes in the proteome profiles of the cells, which affected more than 30% of detected cellular proteins. Using mass spectrometry, we identified 239 proteins affected by dedifferentiation into spheroids as compared to the parental cells. These proteins represented such regulatory processes as transcription, cell cycle regulation, apoptosis, cell adhesion, metabolism, intracellular transport, stress response, and angiogenesis. A number of potent regulators of stemness, differentiation and cancer were detected as subnetworks formed by the identified proteins. Our validation tissue microarray study of 30 neuroblastoma cases confirmed that two of the identified proteins, DISC1 and DNA-PKcs, had their expression increased in advanced malignancies. Thus, our report unveiled extensive changes of the cellular proteome upon dedifferentiation of neuroblastoma cells, indicated top subnetworks and clusters of molecular mechanisms involved in dedifferentiation, and provided candidate biomarkers for clinical studies. (C) 2014 Elsevier Inc. All rights reserved.
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| 8. |
- Woksepp, H., et al.
(författare)
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Proteomics of transforming growth factorβ1(TGFβ1) signaling in 184A1 human breast epithelial cells suggests the involvement of casein kinase2α inTGFβ1-dependent p53 phosphorylation at Ser392
- 2019
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Ingår i: Experimental Oncology. - 0204-3564. ; 41:4, s. 304-311
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Tidskriftsartikel (refereegranskat)abstract
- Aim: Transforming growth factor β1 (TGF β1) is a potent regulator of breast tumorigenesis. It inhibits proliferation of carcinoma cells, but the strength of its inhibitory action varies for cells from benigh, non-metastatic or metastatic tumors. The aim of this work was to generate a proteome profile of TGF β1 action on non-tumorigenic human breast epithelial cells 184A1, and validate predicted involvement of casein kinase 2α (CK2α), p53 and structure-specific recognition protein-1 (SSRP1). Materials and Methods: Two-dimensional gel electrophoresis and mass spectrometry were used to identify TGF β1-regulated proteins in 184A1 human breast immortalized non-tumorigenic cells. 184A1 cells may serve as a model of benign breast neoplasia. These cells were obtained from normal mammary tissue, were immortalized but are not malignant, and were obtained from the American Type Culture Collection. The systemic analysis was performed by using the Cytoscape tool. Transfection of cells with CK2α construct and small interfering RNAs to CK2α and SSRP1 were used to assess an impact of CK2α and SSRP1 on phosphorylation of the p53 and cell proliferation. Results: Proliferation of 184A1 cells was transiently inhibited by TGF β1. We identified 100 and 47 unique proteins which changed their expression and/or 35S-incorporation, respectively, upon treatment with TGF β1 for 2 h, 8 h or 24 h. Cell proliferation, death, migration, and metabolism were among the biological regulatory processes retrieved by the network analysis as affected by the identified proteins. The network analysis suggested that TGF β1 may affect the phosphorylation of p53 at Ser392 by engaging CK2α. This was confirmed by the immunoblotting and cell proliferation assays. Conclusion: We report here the list of 147 TGF β1-regulated proteins in immortalized non-tumorigenic human breast epithelial cells, and show involvement of CK2α in the regulation of p53 Ser392 phosphorylation.
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