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Article ; Online: MOTS-c, the Most Recent Mitochondrial Derived Peptide in Human Aging and Age-Related Diseases.

Mohtashami, Zahra / Singh, Mithalesh K / Salimiaghdam, Nasim / Ozgul, Mustafa / Kenney, M Cristina

International journal of molecular sciences

2022 Volume 23, Issue 19

Abstract: MOTS-c, a 16 amino acid mitochondrial derived peptide, is encoded from the 12S rRNA region of the mitochondrial genome. Under stress conditions, MOTS-c translocates to the nucleus where it regulates a wide range of genes in response to metabolic ... ...

Abstract	MOTS-c, a 16 amino acid mitochondrial derived peptide, is encoded from the 12S rRNA region of the mitochondrial genome. Under stress conditions, MOTS-c translocates to the nucleus where it regulates a wide range of genes in response to metabolic dysfunction. It is colocalized to mitochondria in various tissues and is found in plasma, but the levels decline with age. Since MOTS-c has important cellular functions as well as a possible hormonal role, it has been shown to have beneficial effects on age-related diseases including Diabetes, Cardiovascular diseases, Osteoporosis, postmenopausal obesity and Alzheimer. Aging is characterized by gradual loss of (mitochondrial) metabolic balance, decreased muscle homeostasis and eventual diminished physical capability, which potentially can be reversed with MOTS-c treatment. This review examines the latest findings on biological effects of MOTS-c as a nuclear regulatory peptide and focuses on the role of MOTS-c in aging and age-related disorders, including mechanisms of action and therapeutic potential.
MeSH term(s)	Aging ; Amino Acids/metabolism ; Female ; Humans ; Mitochondria/metabolism ; Mitochondrial Proteins/metabolism ; Peptides/metabolism
Chemical Substances	Amino Acids ; Mitochondrial Proteins ; Peptides
Language	English
Publishing date	2022-10-09
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms231911991
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Article ; Online: Role of dual specificity phosphatases (DUSPs) in melanoma cellular plasticity and drug resistance.

Singh, Mithalesh K / Altameemi, Sarah / Lares, Marcos / Newton, Michael A / Setaluri, Vijayasaradhi

Scientific reports

2022 Volume 12, Issue 1, Page(s) 14395

Abstract: Melanoma cells exhibit phenotypic plasticity that allows transition from a proliferative and differentiated phenotype to a more invasive and undifferentiated or transdifferentiated phenotype often associated with drug resistance. The mechanisms that ... ...

Abstract	Melanoma cells exhibit phenotypic plasticity that allows transition from a proliferative and differentiated phenotype to a more invasive and undifferentiated or transdifferentiated phenotype often associated with drug resistance. The mechanisms that control melanoma phenotype plasticity and its role in drug resistance are not fully understood. We previously demonstrated that emergence of MAPK inhibitor (MAPKi)-resistance phenotype is associated with decreased expression of stem cell proliferation genes and increased expression of MAPK inactivation genes, including dual specificity phosphatases (DUSPs). Several members of the DUSP family genes, specifically DUSP1, -3, -8 and -9, are expressed in primary and metastatic melanoma cell lines and pre-and post BRAFi treated melanoma cells. Here, we show that knockdown of DUSP1 or DUSP8 or treatment with BCI, a pharmacological inhibitor of DUSP1/6 decrease the survival of MAPKi-resistant cells and sensitizes them to BRAFi and MEKi. Pharmacological inhibition of DUSP1/6 upregulated nestin, a neural crest stem cell marker, in both MAPKi-sensitive cells and cells with acquired MAPKi-resistance. In contrast, treatment with BCI resulted in upregulation of MAP2, a neuronal differentiation marker, only in MAPKi-sensitive cells but caused downregulation of both MAP2 and GFAP, a glial marker, in all MAPKi-resistant cell lines. These data suggest that DUSP proteins are involved in the regulation of cellular plasticity cells and melanoma drug resistance and are potential targets for treatment of MAPKi-resistant melanoma.
MeSH term(s)	Cell Line, Tumor ; Cell Plasticity/genetics ; Drug Resistance, Neoplasm/genetics ; Dual-Specificity Phosphatases/genetics ; Humans ; Melanoma/drug therapy ; Melanoma/genetics ; Melanoma/pathology ; Protein Kinase Inhibitors/pharmacology
Chemical Substances	Protein Kinase Inhibitors ; Dual-Specificity Phosphatases (EC 3.1.3.48)
Language	English
Publishing date	2022-08-23
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-022-18578-x
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Article ; Online: Using Human 'Personalized' Cybrids to Identify Drugs/Agents That Can Regulate Chronic Lymphoblastic Leukemia Mitochondrial Dysfunction.

Singh, Lata / Atilano, Shari / Chwa, Marilyn / Singh, Mithalesh K / Ozgul, Mustafa / Nesburn, Anthony / Kenney, M Cristina

International journal of molecular sciences

2023 Volume 24, Issue 13

Abstract: This study uses personalized chronic lymphoblastic leukemia (CLL) cybrid cells to test various drugs/agents designed to improve mitochondrial function and cell longevity. Age-matched control (NL) and CLL cybrids were created. The NL and CLL cybrids were ... ...

Abstract	This study uses personalized chronic lymphoblastic leukemia (CLL) cybrid cells to test various drugs/agents designed to improve mitochondrial function and cell longevity. Age-matched control (NL) and CLL cybrids were created. The NL and CLL cybrids were treated with ibrutinib (Ibr-10 μM), mitochondrial-targeted nutraceuticals such as alpha lipoic acid (ALA-1 mM), amla (Aml-300 μg), melatonin (Mel-1 mM), resveratrol (Res-100 μM) alone, or a combination of ibrutinib with nutraceuticals (Ibr + ALA, Ibr + Aml, Ibr + Mel, or Ibr + Res) for 48 h. MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliumbromide), H2DCFDA(2',7' Dichlorodihydrofluorescein diacetate), and JC1 assays were used to measure the cellular metabolism, intracellular ROS levels, and mitochondrial membrane potential (∆ψm), respectively. The expression levels of genes associated with antioxidant enzymes (
MeSH term(s)	Humans ; Antioxidants/metabolism ; Antioxidants/pharmacology ; Antioxidants/therapeutic use ; Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy ; Leukemia, Lymphocytic, Chronic, B-Cell/metabolism ; Leukemia, Myeloid, Acute/drug therapy ; Leukemia, Myeloid, Acute/metabolism ; Mitochondria/drug effects ; Mitochondria/metabolism ; Mitochondria/pathology ; Reactive Oxygen Species/metabolism ; Drug Resistance, Neoplasm/drug effects ; Hybrid Cells ; Dietary Supplements ; Membrane Potential, Mitochondrial/drug effects ; Gene Expression/drug effects
Chemical Substances	Antioxidants ; Reactive Oxygen Species ; ibrutinib (1X70OSD4VX)
Language	English
Publishing date	2023-07-03
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms241311025
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Article ; Online: Hypoxia-induced transcriptional differences in African and Asian versus European diabetic cybrids.

Dolinko, Andrew H / Chwa, Marilyn / Schneider, Kevin / Singh, Mithalesh K / Atilano, Shari / Wu, Jie / Kenney, M Cristina

Scientific reports

2023 Volume 13, Issue 1, Page(s) 3818

Abstract: Diabetic retinopathy (DR) is the most common diabetic microvascular complication and cause of blindness in adults under the age of 65. Our results suggest that, when comparing transcriptomes of cultures grown in hypoxic conditions versus room-air, ... ...

Abstract	Diabetic retinopathy (DR) is the most common diabetic microvascular complication and cause of blindness in adults under the age of 65. Our results suggest that, when comparing transcriptomes of cultures grown in hypoxic conditions versus room-air, cybrids containing mitochondria from African and Asian diabetic subjects ([Afr + Asi]/DM) have some uniquely different transcriptome profiles compared to European/diabetic (Euro/DM) cybrids (e.g., fatty acid metabolism: EnrichR rank 10 in [Afr + Asi]/DM, rank 85 in Euro/DM; Endocytosis: rank 25 in [Afr + Asi]/DM, rank 5 in Euro/DM; Ubiquitin Mediated Proteolysis: rank 34 in [Afr + Asi]/DM, rank 7 in Euro/DM). As determined by both RNA-seq and qRT-PCR results, transcription of the gene encoding oleoyl-ACP hydrolase (OLAH) was significantly increased in [Afr + Asi]/DM cybrids compared to Euro/DM cybrids in hypoxic conditions. Additionally, our results show that in hypoxic conditions, Euro/DM cybrids and [Afr + Asi]/DM cybrids show similar decreases in ROS production. All cybrids showed decreased ZO1-minus protein levels, but their phagocytic functions were not significantly altered in hypoxic conditions. In conclusion, our findings suggest that the "molecular memory" imparted by [Afr + Asi]/DM mtDNA may act through one of the molecular pathways seen in transcriptome analysis, such as fatty acid metabolism, without significantly changing essential RPE functions.
MeSH term(s)	Adult ; Humans ; Diabetic Retinopathy/genetics ; Asian ; Black People ; Hypoxia/genetics ; Fatty Acids ; Diabetes Mellitus/genetics
Chemical Substances	Fatty Acids
Language	English
Publishing date	2023-03-07
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-30518-x
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Article ; Online: Tubercular DNA PCR of ocular fluids and blood in cases of presumed ocular tuberculosis: a pilot study.

Chawla, Rohan / Singh, Mithalesh K / Singh, Lata / Shah, Pooja / Kashyap, Seema / Azad, Shorya / Venkatesh, Pradeep / Sen, Seema

Therapeutic advances in ophthalmology

2022 Volume 14, Page(s) 25158414221123522

Abstract: Background: The definitive diagnosing of ocular tuberculosis (TB) is difficult; therefore, there is a need of better understanding of investigating TB DNA in presumed ocular TB patients.: Objectives: The aim of this study is to correlate tubercular ... ...

Abstract	Background: The definitive diagnosing of ocular tuberculosis (TB) is difficult; therefore, there is a need of better understanding of investigating TB DNA in presumed ocular TB patients. Objectives: The aim of this study is to correlate tubercular DNA PCR of aqueous/vitreous and blood in cases of presumed ocular TB. Design: A prospective study. Methods: DNA was extracted from aqueous of cases of choroidal tuberculoma (group 1) and serpiginous choroiditis (group 2) and from vitreous of cases of vasculitis (group 3) and macular hole/retinal detachment (group 4). Gel-based PCR and real-time PCR amplification were performed using IS6110 primer on ocular fluids. The same was also performed on the blood samples of cases in which tubercular DNA was detected in the ocular fluids. Results: Overall, 31 cases were analysed in our study. Tubercular DNA was detected in ocular fluids of seven cases: group 1, two cases (67%); group 2, one case (17%); group 3, four cases (27%); and no case of group 4. Blood samples of six of these seven patients were positive for tubercular DNA. Of these six patients, four had evidence of systemic TB and were on ATT. Two cases had no evidence of active systemic TB, yet PCR was positive from blood and ocular fluids. Conclusion: Tubercular DNA detected from ocular fluids may possibly be due to bystander DNA and may not indicate primary ocular tubercular infection. Thus, caution must be exercised prior to labelling a case of uveitis as being tubercular based on the results of molecular assays on ocular fluids alone. The results of PCR on ocular fluids should be correlated with PCR on blood and systemic findings.
Language	English
Publishing date	2022-09-18
Publishing country	United States
Document type	Journal Article
ISSN	2515-8414
ISSN (online)	2515-8414
DOI	10.1177/25158414221123522
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Article ; Online: Differential Epigenetic Status and Responses to Stressors between Retinal Cybrids Cells with African versus European Mitochondrial DNA: Insights into Disease Susceptibilities.

Atilano, Shari R / Abedi, Sina / Ianopol, Narcisa V / Singh, Mithalesh K / Norman, J Lucas / Malik, Deepika / Falatoonzadeh, Payam / Chwa, Marilyn / Nesburn, Anthony B / Kuppermann, Baruch D / Kenney, M Cristina

Cells

2022 Volume 11, Issue 17

Abstract: Mitochondrial (mt) DNA can be classified into haplogroups, which represent populations with different geographic origins. Individuals of maternal African backgrounds (L haplogroup) are more prone to develop specific diseases compared those with maternal ... ...

Abstract	Mitochondrial (mt) DNA can be classified into haplogroups, which represent populations with different geographic origins. Individuals of maternal African backgrounds (L haplogroup) are more prone to develop specific diseases compared those with maternal European-H haplogroups. Using a cybrid model, effects of amyloid-β (Amyβ), sub-lethal ultraviolet (UV) radiation, and 5-Aza-2'-deoxycytidine (5-aza-dC), a methylation inhibitor, were investigated. Amyβ treatment decreased cell metabolism and increased levels of reactive oxygen species in European-H and African-L cybrids, but lower mitochondrial membrane potential (ΔΨM) was found only in African-L cybrids. Sub-lethal UV radiation induced higher expression levels of
MeSH term(s)	DNA, Mitochondrial/genetics ; DNA, Mitochondrial/metabolism ; Disease Susceptibility/metabolism ; Epigenesis, Genetic ; Extracellular Matrix Proteins/metabolism ; Humans ; Mitochondria/genetics ; Mitochondria/metabolism ; Polymorphism, Single Nucleotide ; Ubiquitin Thiolesterase/metabolism
Chemical Substances	DNA, Mitochondrial ; EFEMP1 protein, human ; Extracellular Matrix Proteins ; USP25 protein, human ; Ubiquitin Thiolesterase (EC 3.4.19.12)
Language	English
Publishing date	2022-08-26
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells11172655
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Article ; Online: Impacts of Bacteriostatic and Bactericidal Antibiotics on the Mitochondria of the Age-Related Macular Degeneration Cybrid Cell Lines.

Salimiaghdam, Nasim / Singh, Lata / Singh, Mithalesh K / Chwa, Marilyn / Atilano, Shari R / Mohtashami, Zahra / Nesburn, Anthony B / Kuppermann, Baruch D / Lu, Stephanie Y / Kenney, M Cristina

Biomolecules

2022 Volume 12, Issue 5

Abstract: We assessed the potential negative effects of bacteriostatic and bactericidal antibiotics on the AMD cybrid cell lines (K, U and J haplogroups). AMD cybrid cells were created and cultured in 96-well plates and treated with tetracycline (TETRA) and ... ...

Abstract	We assessed the potential negative effects of bacteriostatic and bactericidal antibiotics on the AMD cybrid cell lines (K, U and J haplogroups). AMD cybrid cells were created and cultured in 96-well plates and treated with tetracycline (TETRA) and ciprofloxacin (CPFX) for 24 h. Reactive oxygen species (ROS) levels, mitochondrial membrane potential (ΔψM), cellular metabolism and ratio of apoptotic cells were measured using H2DCFDA, JC1, MTT and flow cytometry assays, respectively. Expression of genes of antioxidant enzymes, and pro-inflammatory and pro-apoptotic pathways were evaluated by quantitative real-time PCR (qRT-PCR). Higher ROS levels were found in U haplogroup cybrids when treated with CPFX 60 µg/mL concentrations, lower ΔψM of all haplogroups by CPFX 120 µg/mL, diminished cellular metabolism in all cybrids with CPFX 120 µg/mL, and higher ratio of dead cells in K and J cybrids. CPFX 120 µg/mL induced overexpression of
MeSH term(s)	Anti-Bacterial Agents/metabolism ; Anti-Bacterial Agents/pharmacology ; Cell Line ; Ciprofloxacin/pharmacology ; Humans ; Interleukin-6/metabolism ; Macular Degeneration/drug therapy ; Macular Degeneration/genetics ; Macular Degeneration/metabolism ; Mitochondria/metabolism ; Reactive Oxygen Species/metabolism ; Tetracyclines
Chemical Substances	Anti-Bacterial Agents ; Interleukin-6 ; Reactive Oxygen Species ; Tetracyclines ; Ciprofloxacin (5E8K9I0O4U)
Language	English
Publishing date	2022-05-07
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2701262-1
ISSN	2218-273X ; 2218-273X
ISSN (online)	2218-273X
ISSN	2218-273X
DOI	10.3390/biom12050675
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Article ; Online: Altered Retrograde Signaling Patterns in Breast Cancer Cells Cybrids with H and J Mitochondrial DNA Haplogroups.

Chang, Steven / Singh, Lata / Thaker, Kunal / Abedi, Sina / Singh, Mithalesh K / Patel, Tej H / Chwa, Marilyn / Atilano, Shari R / Udar, Nitin / Bota, Daniela / Kenney, Maria Cristina

International journal of molecular sciences

2022 Volume 23, Issue 12

Abstract: The aim of this study was to determine the role of retrograde signaling (mitochondria to nucleus) in MCF7 breast cancer cells. Therefore, in the present study, MCF7-H and MCF7-J cybrids were produced using the mitochondria from the same H and J ... ...

Abstract	The aim of this study was to determine the role of retrograde signaling (mitochondria to nucleus) in MCF7 breast cancer cells. Therefore, in the present study, MCF7-H and MCF7-J cybrids were produced using the mitochondria from the same H and J individuals that were already used in our non-diseased retinal pigment epithelium (ARPE19) cybrids. MCF7 cybrids were treated with cisplatin and analyzed for cell viability, mitochondrial membrane potential, ROS, and expression levels of genes associated with the cGAS-STING and cancer-related pathways. Results showed that unlike the ARPE19-H and ARPE19-J cybrids, the untreated MCF7-H and MCF7-J cybrids had similar levels of ATP, lactate, and OCR: ECAR ratios. After cisplatin treatment, MCF7-H and MCF7-J cybrids showed similar (
MeSH term(s)	Breast Neoplasms/genetics ; Breast Neoplasms/metabolism ; Cisplatin/metabolism ; Cisplatin/pharmacology ; DNA, Mitochondrial/genetics ; DNA, Mitochondrial/metabolism ; Female ; Humans ; Mitochondria/genetics ; Mitochondria/metabolism ; Nucleotidyltransferases/metabolism ; Reactive Oxygen Species/metabolism
Chemical Substances	DNA, Mitochondrial ; Reactive Oxygen Species ; Nucleotidyltransferases (EC 2.7.7.-) ; Cisplatin (Q20Q21Q62J)
Language	English
Publishing date	2022-06-15
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms23126687
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Article ; Online: EPAC Regulates Melanoma Growth by Stimulating mTORC1 Signaling and Loss of EPAC Signaling Dependence Correlates with Melanoma Progression.

Krishnan, Aishwarya / Bhasker, Aishwarya I / Singh, Mithalesh K / Rodriguez, Carlos I / Pérez, Edgardo Castro / Altameemi, Sarah / Lares, Marcos / Khan, Hamidullah / Ndiaye, Mary / Ahmad, Nihal / Schieke, Stefan M / Setaluri, Vijayasaradhi

Molecular cancer research : MCR

2022 Volume 20, Issue 10, Page(s) 1548–1560

Abstract: Exchange proteins directly activated by cAMP (EPAC) belong to a family of RAP guanine nucleotide exchange factors (RAPGEF). EPAC1/2 (RAPGEF3/4) activates RAP1 and the alternative cAMP signaling pathway. We previously showed that the differential growth ... ...

Abstract	Exchange proteins directly activated by cAMP (EPAC) belong to a family of RAP guanine nucleotide exchange factors (RAPGEF). EPAC1/2 (RAPGEF3/4) activates RAP1 and the alternative cAMP signaling pathway. We previously showed that the differential growth response of primary and metastatic melanoma cells to cAMP is mediated by EPAC. However, the mechanisms responsible for this differential response to EPAC signaling are not understood. In this study, we show that pharmacologic inhibition or siRNA-mediated knockdown of EPAC selectively inhibits the growth and survival of primary melanoma cells by downregulation of cell-cycle proteins and inhibiting the cell-cycle progression independent of ERK1/2 phosphorylation. EPAC inhibition results in upregulation of AKT phosphorylation but a downregulation of mTORC1 activity and its downstream effectors. We also show that EPAC regulates both glycolysis and oxidative phosphorylation, and production of mitochondrial reactive oxygen species, preferentially in primary melanoma cells. Employing a series of genetically matched primary and lymph node metastatic (LNM) melanoma cells, and distant organ metastatic melanoma cells, we show that the LNM and metastatic melanoma cells become progressively less responsive and refractory to EPAC inhibition suggesting loss of dependency on EPAC signaling correlates with melanoma progression. Analysis of The Cancer Genome Atlas dataset showed that lower RAPGEF3, RAPGEF4 mRNA expression in primary tumor is a predictor of better disease-free survival of patients diagnosed with primary melanoma suggesting that EPAC signaling facilitates tumor progression and EPAC is a useful prognostic marker. These data highlight EPAC signaling as a potential target for prevention of melanoma progression. Implications: This study establishes loss of dependency on EPAC-mTORC1 signaling as hallmark of primary melanoma evolution and targeting this escape mechanism is a promising strategy for metastatic melanoma.
MeSH term(s)	Guanine Nucleotide Exchange Factors/genetics ; Guanine Nucleotide Exchange Factors/metabolism ; Humans ; Mechanistic Target of Rapamycin Complex 1/genetics ; Mechanistic Target of Rapamycin Complex 1/metabolism ; Melanoma/pathology ; Proto-Oncogene Proteins c-akt/metabolism ; RNA, Messenger/genetics ; RNA, Small Interfering ; Reactive Oxygen Species ; Signal Transduction
Chemical Substances	Guanine Nucleotide Exchange Factors ; RAPGEF4 protein, human ; RNA, Messenger ; RNA, Small Interfering ; Reactive Oxygen Species ; Mechanistic Target of Rapamycin Complex 1 (EC 2.7.11.1) ; Proto-Oncogene Proteins c-akt (EC 2.7.11.1)
Language	English
Publishing date	2022-07-14
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2098788-2
ISSN	1557-3125 ; 1541-7786
ISSN (online)	1557-3125
ISSN	1541-7786
DOI	10.1158/1541-7786.MCR-22-0026
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Article ; Online: Prognostic significance of polo-like kinases in retinoblastoma: correlation with patient outcome, clinical and histopathological parameters.

Singh, Lata / Pushker, Neelam / Sen, Seema / Singh, Mithalesh K / Chauhan, Feeroj A / Kashyap, Seema

Clinical & experimental ophthalmology

2015 Volume 43, Issue 6, Page(s) 550–557

Abstract: Background: Retinoblastoma is evolving, but it is still a therapeutic challenge for pediatric oncologists. Polo-like kinases (PLKs) plays an important role in cell cycle events. They play a crucial role in cell proliferation which may lead to tumour ... ...

Abstract	Background: Retinoblastoma is evolving, but it is still a therapeutic challenge for pediatric oncologists. Polo-like kinases (PLKs) plays an important role in cell cycle events. They play a crucial role in cell proliferation which may lead to tumour formation. The objective of this study is to investigate the role of PLK1 and PLK3 proteins in human retinoblastoma tissues. Design: Non-randomized, prospective study was performed in the Dr R. P. Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India. Participants: This study included 74 primary enucleated retinoblastoma tissues. Methods: Expression of PLK1 and PLK3 protein were assessed in primary enucleated retinoblastoma tissues by immunohistochemistry and western blotting. Main outcome measures: Expression of PLK1 and PLK3 protein were correlated with clinical and histopathological parameters, tumour staging and overall survival of patients. Results: Immunohistochemical results revealed expression of PLK1 in 47/74 (63.51%) cases and PLK3 in 31/74 (41.89%) cases. Western blotting confirmed the immunoreactivity results. Expression of PLK1 showed correlation with poor differentiation and tumour invasion. In addition, PLK1 was statistically significant with massive choroidal invasion, whereas PLK3 did not correlate with any of the clinical or histopathological parameters. There was no statistical correlation in the overall survival of patients with PLK1 and PLK3 expression. Conclusions: PLK1 expression was associated with poor tumour differentiation and histopathological high-risk factors. These proteins may be involved in tumorigenesis and progression of disease. These results suggest that PLK1 may act as a potential therapeutic target and a promising marker for developing potent small molecule inhibitors of PLK isoforms in retinoblastoma.
MeSH term(s)	Blotting, Western ; Cell Cycle Proteins/metabolism ; Child ; Child, Preschool ; Eye Enucleation ; Female ; Humans ; Immunoenzyme Techniques ; Infant ; Male ; Prognosis ; Prospective Studies ; Protein Serine-Threonine Kinases/metabolism ; Proto-Oncogene Proteins/metabolism ; Retinal Neoplasms/enzymology ; Retinal Neoplasms/pathology ; Retinoblastoma/enzymology ; Retinoblastoma/pathology ; Tumor Suppressor Proteins ; Polo-Like Kinase 1
Chemical Substances	Cell Cycle Proteins ; Proto-Oncogene Proteins ; Tumor Suppressor Proteins ; PLK3 protein, human (EC 2.7.1.-) ; Protein Serine-Threonine Kinases (EC 2.7.11.1)
Language	English
Publishing date	2015-05-13
Publishing country	Australia
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2014008-3
ISSN	1442-9071 ; 1442-6404
ISSN (online)	1442-9071
ISSN	1442-6404
DOI	10.1111/ceo.12517
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