Parathyroid Hormone Stimulates Colony Formation of Chick Embryo Chondrocytes in Soft Agar

1989 ◽  
pp. 429-433
Author(s):  
Tatsuya Koike ◽  
Yukio Kato ◽  
Masahiro Iwamoto ◽  
Fujio Suzuki
Keyword(s):  
Parathyroid Hormone ◽  
Chick Embryo ◽  
Colony Formation ◽  
Soft Agar

1α,25-Dihydroxyvitamin D3 stimulates colony formation of chick embryo chondrocytes in soft agar

1990 ◽  
Vol 187 (2) ◽  
pp. 335-338 ◽  
Author(s):  
Katsuhiko Sato ◽  
Masahiro Iwamoto ◽  
Kazuhisa Nakashima ◽  
Fujio Suzuki ◽  
Yukio Kato
Keyword(s):  
Chick Embryo ◽  
Colony Formation ◽  
Soft Agar ◽  
Dihydroxyvitamin D3

scholarly journals Functional consequences of the first reported mutations of the proto-oncogene PTTG1IP/PBF

2017 ◽  
Vol 24 (9) ◽  
pp. 459-474 ◽  
Author(s):  
W Imruetaicharoenchoke ◽  
A Fletcher ◽  
W Lu ◽  
R J Watkins ◽  
B Modasia ◽  
...  
Keyword(s):  
Colony Formation ◽  
Human Cancer ◽  
Soft Agar ◽  
Cellular Migration ◽  
3T3 Fibroblasts ◽  
Wide Range ◽  
Functional Consequences ◽  
Altered Cell ◽  
Proliferation Assays ◽  
Nih 3T3

Pituitary tumor-transforming gene 1-binding factor (PTTG1IP; PBF) is a multifunctional glycoprotein, which is overexpressed in a wide range of tumours, and significantly associated with poorer oncological outcomes, such as early tumour recurrence, distant metastasis, extramural vascular invasion and decreased disease-specific survival. PBF transforms NIH 3T3 fibroblasts and induces tumours in nude mice, while mice harbouring transgenic thyroidal PBF expression show hyperplasia and macrofollicular lesions. Our assumption that PBF becomes an oncogene purely through increased expression has been challenged by the recent report of mutations in PBF within the Catalogue of Somatic Mutations in Cancer (COSMIC) database. We therefore sought to determine whether the first 10 PBF missense substitutions in human cancer might be oncogenic. Anisomycin half-life studies revealed that most mutations were associated with reduced protein stability compared to wild-type (WT) PBF. Proliferation assays narrowed our interest to two mutational events which significantly altered cell turnover: C51R and R140W. C51R was mainly confined to the endoplasmic reticulum while R140W was apparent in the Golgi apparatus. Both C51R and R140W lost the capacity to induce cellular migration and significantly reduced cell invasion. Colony formation and soft agar assays demonstrated that, in contrast to WT PBF, both mutants were unable to elicit significant colony formation or anchorage-independent growth. However, C51R and R140W retained the ability to repress radioiodide uptake, a functional hallmark of PBF. Our data reveal new insight into PBF function and confirm that, rather than being oncogenic, mutations in PBF are likely to be passenger effects, with overexpression of PBF the more important aetiological event in human cancer.

The Src family kinase Fgr is a transforming oncoprotein that functions independently of SH3-SH2 domain regulation

2018 ◽  
Vol 11 (553) ◽  
pp. eaat5916 ◽  
Author(s):  
Kexin Shen ◽  
Jamie A. Moroco ◽  
Ravi K. Patel ◽  
Haibin Shi ◽  
John R. Engen ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Colony Formation ◽  
Family Members ◽  
Cellular Transformation ◽  
Soft Agar ◽  
Kinase Domain ◽  
Sh2 Domain ◽  
Mass Spectrometry Data ◽  
Activation Loop ◽  
Exchange Mass Spectrometry

Fgr is a member of the Src family of nonreceptor tyrosine kinases, which are overexpressed and constitutively active in many human cancers. Fgr expression is restricted to myeloid hematopoietic cells and is markedly increased in a subset of bone marrow samples from patients with acute myeloid leukemia (AML). Here, we investigated the oncogenic potential of Fgr using Rat-2 fibroblasts that do not express the kinase. Expression of either wild-type or regulatory tail-mutant constructs of Fgr promoted cellular transformation (inferred from colony formation in soft agar), which was accompanied by phosphorylation of the Fgr activation loop, suggesting that the kinase domain of Fgr functions independently of regulation by its noncatalytic SH3-SH2 region. Unlike other family members, recombinant Fgr was not activated by SH3-SH2 domain ligands. However, hydrogen-deuterium exchange mass spectrometry data suggested that the regulatory SH3 and SH2 domains packed against the back of the kinase domain in a Src-like manner. Sequence alignment showed that the activation loop of Fgr was distinct from that of all other Src family members, with proline rather than alanine at the +2 position relative to the activation loop tyrosine. Substitution of the activation loop of Fgr with the sequence from Src partially inhibited kinase activity and suppressed colony formation. Last, Fgr expression enhanced the sensitivity of human myeloid progenitor cells to the cytokine GM-CSF. Because its kinase domain is not sensitive to SH3-SH2–mediated control, simple overexpression of Fgr without mutation may contribute to oncogenic transformation in AML and other blood cancers.

scholarly journals The Soft Agar Colony Formation Assay

10.3791/51998 ◽  
2014 ◽  
Author(s):  
Stanley Borowicz ◽  
Michelle Van Scoyk ◽  
Sreedevi Avasarala ◽  
Manoj Kumar Karuppusamy Rathinam ◽  
Jordi Tauler ◽  
...  
Keyword(s):  
Colony Formation ◽  
Soft Agar ◽  
Colony Formation Assay

scholarly journals Three New Steroid Biglycosides, Plancisides A, B, and C, from the Starfish Acanthaster planci

2014 ◽  
Vol 9 (9) ◽  
pp. 1934578X1400900 ◽  
Author(s):  
Alla A. Kicha ◽  
Thi H. Dinh ◽  
Natalia V. Ivanchina ◽  
Timofey V. Malyarenko ◽  
Anatoly I. Kalinovsky ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Lines ◽  
Colony Formation ◽  
Clonogenic Assay ◽  
Ethanolic Extract ◽  
Soft Agar ◽  
Steroid Glycoside ◽  
Acanthaster Planci ◽  
Hct 116 ◽  
Soft Agar Clonogenic Assay

Three new steroid biglycosides, plancisides A-C (1-3), were isolated from the ethanolic extract of the starfish Acanthaster planci. The structures of 1-3 were determined by extensive NMR and ESI-MS techniques, as (24 S)-28- O-[β-D-galactofuranosyl-(1→5)-α-L-arabinofuranosyl]-24-methyl-5α-cholestane-3β,4β,6α,8,15β,16β,28-heptol (1), (24 S)-28- O-[α-L-fucopyranosyl-(1→2)-3- O-methyl-β-D-xylopyranosyl]–24-methyl-5α-cholestane-3β,4β,6α,8,15β,16β,28-heptol (2) and (24 S)-28- O-[2,4-di- O-methyl-β-D-xylopyranosyl-(1→2)-α-L-arabinofuranosyl]-24-methyl-5α-cholestane-3β,4β,6α,8,15β,16β,28-heptol 6- O-sulfate (3), respectively. Compound 2 is the first steroid glycoside containing an α-fucopyranose unit found from starfish. Compound 1 slightly inhibits cell proliferation of HCT-116, T-47D, and RPMI-7951 cancer cell lines, but has no effect on colony formation of these cells in a soft agar clonogenic assay.

scholarly journals Human myeloid progenitor cells expressing HLA antigens

Blood ◽  
1979 ◽  
Vol 53 (4) ◽  
pp. 794-797 ◽  
Author(s):  
JH Fitchen ◽  
MJ Cline
Keyword(s):  
Progenitor Cells ◽  
Indirect Effect ◽  
Colony Formation ◽  
Hla Antigens ◽  
Soft Agar ◽  
Myeloid Progenitor ◽  
Hla Type ◽  
Myeloid Progenitor Cells ◽  
Marrow Cells

Abstract Marrow cells of known HLA type were incubated with HLA antiserum plus complement and then plated in soft agar. Colony formation was consistently inhibited by appropriate HLA antisera. Mixing experiments excluded an indirect effect on CFU-C by lysis of mature leukocytes. We conclude that human CFU-C express HLA antigens.

scholarly journals NR4A1 Promotes PDGF-BB-Induced Cell Colony Formation in Soft Agar

PLoS ONE ◽  
2014 ◽  
Vol 9 (9) ◽  
pp. e109047 ◽  
Author(s):  
Glenda Eger ◽  
Natalia Papadopoulos ◽  
Johan Lennartsson ◽  
Carl-Henrik Heldin
Keyword(s):  
Colony Formation ◽  
Soft Agar ◽  
Cell Colony

scholarly journals Onconase Restores Cytotoxicity in Dabrafenib-Resistant A375 Human Melanoma Cells and Affects Cell Migration, Invasion and Colony Formation Capability

2019 ◽  
Vol 20 (23) ◽  
pp. 5980 ◽  
Author(s):  
Raineri ◽  
Fasoli ◽  
Campagnari ◽  
Gotte ◽  
Menegazzi
Keyword(s):  
Colony Formation ◽  
Apoptotic Cell ◽  
Gelatin Zymography ◽  
Metastatic Potential ◽  
Soft Agar ◽  
Cell Subpopulation ◽  
Migration And Invasion ◽  
Dependent Manner ◽  
Cell Subpopulations ◽  
Resistant Cells

Melanoma is a lethal tumor because of its severe metastatic potential, and serine/threonine-protein kinase B-raf inhibitors (BRAFi) are used in patients harboring BRAF-mutation. Unfortunately, BRAFi induce resistance. Therefore, we tested the activity of onconase (ONC), a cytotoxic RNase variant, against BRAFi-resistant cells to re-establish the efficacy of the chemotherapy. To do so, an A375 dabrafenib-resistant (A375DR) melanoma cell subpopulation was selected and its behavior compared with that of parental (A375P) cells by crystal violet, 5-Bromo-2’-deoxyuridine incorporation, and cleaved poly(ADP-ribose) polymerase 1 (PARP1) western blot measurements. Then, nuclear p65 Nuclear Factor kappaB (NF-κB) and IκB kinases-α/β (IKK) phosphorylation levels were measured. Gelatin zymography was performed to evaluate metalloproteinase 2 (MMP2) activity. In addition, assays to measure migration, invasion and soft agar colony formation were performed to examine the tumor cell dissemination propensity. ONC affected the total viability and the proliferation rate of both A375P and A375DR cell subpopulations in a dose-dependent manner and also induced apoptotic cell death. Among its pleiotropic effects, ONC reduced nuclear p65 NF-κB amount and IKK phosphorylation level, as well as MMP2 activity in both cell subpopulations. ONC decreased cell colony formation, migration, and invasion capability. Notably, it induced apoptosis and inhibited colony formation and invasiveness more extensively in A375DR than in A375P cells. In conclusion, ONC successfully counteracts melanoma malignancy especially in BRAFi-resistant cells and could become a tool against melanoma recurrence.

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