scholarly journals Oncogenic transformation of NIH/3T3 cells by the overexpression of L-type amino acid transporter 1, a promising anti-cancer target

Oncotarget ◽  
2021 ◽  
Author(s):  
Natsumi Hayashi ◽  
Akitaka Yamasaki ◽  
Shiho Ueda ◽  
Shogo Okazaki ◽  
Yoshiya Ohno ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Transporter ◽  
Oncogenic Transformation ◽  
3T3 Cells ◽  
Anti Cancer ◽  
Acid Transporter ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

Oncogenic Transformation of NIH 3T3 Fibroblasts by BCR-ABL Oncoprotein Requires the IL-3 Receptor.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3370-3370
Author(s):  
Wenjing Tao ◽  
Hui Lin ◽  
Tong Sun ◽  
Ajoy K. Samanta ◽  
Ralph B. Arlinghaus
Keyword(s):  
Kinase Inhibitor ◽  
Philadelphia Chromosome ◽  
Cell System ◽  
Receptor Expression ◽  
Oncogenic Transformation ◽  
3T3 Cells ◽  
Lymphoid Leukemia ◽  
3T3 Fibroblasts ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

Abstract Bcr-Abl is a leukemia-inducing protein, which causes oncogenic transformation of myeloid progenitors in Philadelphia chromosome (Ph)-positive chronic myeloid leukemia (CML) and lymphoid progenitors in Ph+ acute lymphoid leukemia (ALL). Oncogenic transformation of hematopoietic cells by the Bcr-Abl oncoprotein directly involves the activation Jak2 tyrosine kinase and the Stat5 transcription factor. Both proteins are normally linked to the IL-3/GM-CSF receptors for growth and survival. Since fibroblastic cells are not targets of BCR-ABL induced oncogenesis, we determined whether forced expression of the IL-3 receptor would allow oncogenic transformation of NIH 3T3 fibroblasts known to be resistant to transformation by BCR-ABL. NIH 3T3 cells transduced with the human IL-3 receptor a and b chains were highly susceptible to oncogenic transformation by expression of BCR-ABL. Forced expression of both receptor chains but not either one alone allowed efficient foci formation of NIH 3T3 cells expressing BCR-ABL, and these cells formed colonies in soft agar whereas BCR-ABL positive NIH 3T3 cells lacking IL-3 receptor expression did not. The Bcr-Abl kinase inhibitor imatinib mesylate (1 mM) and the Jak kinase inhibitor AG490 (10 mM) strongly inhibited agar colony formation. A small molecule inhibitor of Jak2 kinase, 1,2,3,4,5,6-hexabromocyclohexane reported to be specific for Jak2 (Sandberg et al. J. Med. Chem, 2005)-significantly reduced the phosphorylation of Gab2 at the YxxM motif, which is needed for activation of the PI-3 kinase and Akt, two proteins that are part of the Bcr-Abl/Jak2 Network (Samanta et al. Cancer Res, 2006). These findings indicate that Bcr-Abl oncoprotein requires the IL-3 receptor/Jak2/Stat5 pathways for oncogenic transformation of NIH 3T3 fibroblasts, and may explain partially why Bcr-Abl oncogenesis is restricted to hematopoietic malignancies. Furthermore, this cell system in fibroblastic and other cell lineages will provide a model to probe the detailed steps that require IL-3 receptor and Jak2 for Bcr-Abl induced leukemia.

scholarly journals The Fn14 cytoplasmic tail binds tumour-necrosis-factor-receptor-associated factors 1, 2, 3 and 5 and mediates nuclear factor-kappaB activation

2003 ◽  
Vol 371 (2) ◽  
pp. 395-403 ◽  
Author(s):  
Sharron A.N. BROWN ◽  
Christine M. RICHARDS ◽  
Heather N. HANSCOM ◽  
Sheau-Line Y. FENG ◽  
Jeffrey A. WINKLES
Keyword(s):  
Amino Acid ◽  
Tumour Necrosis Factor ◽  
Tumour Necrosis ◽  
Cytoplasmic Tail ◽  
Luciferase Reporter ◽  
Nuclear Factor ◽  
3T3 Cells ◽  
Nih 3T3 ◽  
Necrosis Factor ◽  
Nih 3T3 Cells

Fn14 is a growth-factor-inducible immediate-early-response gene encoding a 102-amino-acid type I transmembrane protein. The human Fn14 protein was recently identified as a cell-surface receptor for the tumour necrosis factor (TNF) superfamily member named TWEAK (TNF-like weak inducer of apoptosis). In the present paper, we report that the human TWEAK extracellular domain can also bind the murine Fn14 protein. Furthermore, site-specific mutagenesis and directed yeast two-hybrid interaction assays revealed that the TNFR-associated factor (TRAF) 1, 2, 3 and 5 adaptor molecules bind the murine Fn14 cytoplasmic tail at an overlapping, but non-identical, amino acid sequence motif. We also found that TWEAK treatment of quiescent NIH 3T3 cells stimulates inhibitory κBα phosphorylation and transcriptional activation of a nuclear factor-κB (NF-κB) enhancer/luciferase reporter construct. Fn14 overexpression in transiently transfected NIH 3T3 cells also promotes NF-κB activation, and this cellular response requires an intact TRAF binding site. These results indicate that Fn14 is a functional TWEAK receptor that can associate with four distinct TRAF family members and stimulate the NF-κB transcription factor signalling pathway.

scholarly journals Inhibition of v-src-induced transformation by a GTPase-activating protein

1991 ◽  
Vol 11 (5) ◽  
pp. 2812-2818 ◽  
Author(s):  
M Nori ◽  
U S Vogel ◽  
J B Gibbs ◽  
M J Weber
Keyword(s):  
Active Form ◽  
Signalling Pathway ◽  
Oncogenic Transformation ◽  
Gtpase Activating Protein ◽  
3T3 Cells ◽  
Inactive Form ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

Previous work has shown that microinjection into cells of antibodies against p21ras blocks transformation by src, suggesting that oncogenic transformation by pp60v-src is dependent on p21ras. The activity of p21ras itself is regulated by its cyclic association with GDP-GTP, where p21ras-GTP is the active form and p21ras-GDP is the inactive form. A GTPase-activating protein (GAP) mediates the inactivation of p21ras by facilitating the conversion of the active p21ras-GTP to the inactive p21ras-GDP. This predicts that overexpression of GAP would inactivate p21ras and block transformation of cells by src. In this paper, we confirm this prediction. We report that overexpression of GAP in NIH 3T3 cells blocks transformation by pp60v-src but not by v-ras. Susceptibility to transformation by v-src is restored when GAP expression is lowered to levels comparable to that in control cells. These results support the suggestion that p21ras plays a central role in the signalling pathway used by pp60v-src.

Cell-dependent efficiency of reiterated nuclear signals in a mutant simian virus 40 oncoprotein targeted to the nucleus

1988 ◽  
Vol 8 (12) ◽  
pp. 5495-5503
Author(s):  
L Fischer-Fantuzzi ◽  
C Vesco
Keyword(s):  
Amino Acid ◽  
Nuclear Transport ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Large T Antigen ◽  
3T3 Cells ◽  
Rat Embryo ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

We investigated the requisites for, and functional consequences of, the relocation to the nucleus of a transforming nonkaryophilic mutant of the simian virus 40 large T antigen (a natural deletion mutant lacking an internal large-T-antigen domain that includes the signal for nuclear transport). Synthetic oligonucleotides were used to obtain gene variants with one or more copies of the signal-specifying sequence inserted near the gene 3' end, in a region dispensable for the main large-T-antigen functions. The analysis of stable transfectant populations showed that mouse NIH 3T3 cells, rat embryo fibroblasts, and simian CS cells (a subclone of CV1 cells) differed considerably in their ability to localize some variant molecules into the nucleus. CS cells were always the most efficient, and NIH 3T3 cells were the least efficient. The nuclear localization improved either with reiteration of the signal or with a left-flank modification of the signal amino acid context. Three signals appeared to be necessary and sufficient, even in NIH 3T3 cells, to obtain a nuclear accumulation comparable to that of wild-type simian virus 40 large T antigen; other signal-cell combinations caused a large variability in subcellular localization among cells of the same population, as if the nuclear uptake of some molecules depended on individual cell states. The effect of the modified location on the competence of the protein to alter cell growth was examined by comparing the activity of variants containing either the normal signal or a signal with a mutation (corresponding to large-T-antigen amino acid 128) that prevented nuclear transport. It was found that the nuclear variant was slightly more active than the cytoplasmic variants in rat embryo fibroblasts and NIH 3T3 cells and was notably less active in CS cells.

Volume regulation in NIH/3T3 cells not expressing P-glycoprotein. II. Chloride and amino acid fluxes

1997 ◽  
Vol 272 (6) ◽  
pp. C1804-C1809 ◽  
Author(s):  
J. Moran ◽  
D. Miranda ◽  
C. Pena-Segura ◽  
H. Pasantes-Morales
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Regulatory Volume Decrease ◽  
Disulfonic Acid ◽  
Channel Blockers ◽  
3T3 Cells ◽  
P Glycoprotein ◽  
Arachidonic Acids ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

The osmolyte function of amino acids and Cl in native NIH/3T3 cells not expressing the P-glycoprotein was examined by investigating the free amino acid concentration and the swelling-activated efflux of [3H]taurine, as representative of amino acids, and of 125I, as a tracer for Cl. Taurine and 125I efflux was activated by 20 and 30% hyposmotic solutions. At 50% hyposmotic solutions, the osmolyte pool was essentially depleted. The Cl channel blockers 5-nitro-2-(3-phenylpropyl-amino)benzoic acid, 1,9-dideoxyforskolin, dipyridamole, and niflumic acid inhibited the release of the two osmolytes by 80-95%. 4,4'-Diisothiocyanostilbene-2,2'-disulfonic acid (400 microM) decreased the efflux of taurine 80% without affecting that of 125I. Linolenic and arachidonic acids (5-20 microM) showed a concentration-dependent inhibitory effect on taurine and 125I fluxes. Omission of Ca decreased osmolyte fluxes by 16%. Verapamil inhibited the osmolyte release only at 500 microM. Nimodipine at 25 and 50 microM decreased the release of [3H]taurine and 125I by approximately 60 and 80%, respectively, but this effect was independent of the presence of extracellular Ca. These results indicate that amino acids and Cl function as osmolytes during regulatory volume decrease in native NIH/ 3T3 cells.

scholarly journals JPH203, a newly developed anti-cancer drug, shows a preincubation inhibitory effect on L-type amino acid transporter 1 function

2020 ◽  
Vol 144 (1) ◽  
pp. 16-22 ◽  
Author(s):  
Kentaro Okunushi ◽  
Tomomi Furihata ◽  
Hanae Morio ◽  
Yasuhide Muto ◽  
Kosuke Higuchi ◽  
...  
Keyword(s):  
Amino Acid ◽  
Inhibitory Effect ◽  
Amino Acid Transporter ◽  
Cancer Drug ◽  
Anti Cancer ◽  
Acid Transporter

scholarly journals Inhibition of v-src-induced transformation by a GTPase-activating protein.

1991 ◽  
Vol 11 (5) ◽  
pp. 2812-2818 ◽  
Author(s):  
M Nori ◽  
U S Vogel ◽  
J B Gibbs ◽  
M J Weber
Keyword(s):  
Active Form ◽  
Signalling Pathway ◽  
Oncogenic Transformation ◽  
Gtpase Activating Protein ◽  
3T3 Cells ◽  
Inactive Form ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

Previous work has shown that microinjection into cells of antibodies against p21ras blocks transformation by src, suggesting that oncogenic transformation by pp60v-src is dependent on p21ras. The activity of p21ras itself is regulated by its cyclic association with GDP-GTP, where p21ras-GTP is the active form and p21ras-GDP is the inactive form. A GTPase-activating protein (GAP) mediates the inactivation of p21ras by facilitating the conversion of the active p21ras-GTP to the inactive p21ras-GDP. This predicts that overexpression of GAP would inactivate p21ras and block transformation of cells by src. In this paper, we confirm this prediction. We report that overexpression of GAP in NIH 3T3 cells blocks transformation by pp60v-src but not by v-ras. Susceptibility to transformation by v-src is restored when GAP expression is lowered to levels comparable to that in control cells. These results support the suggestion that p21ras plays a central role in the signalling pathway used by pp60v-src.

scholarly journals Resistance to oncogenic transformation in revertant R1 of human ras-transformed NIH 3T3 cells.

1989 ◽  
Vol 9 (5) ◽  
pp. 2258-2263 ◽  
Author(s):  
N Kuzumaki ◽  
Y Ogiso ◽  
A Oda ◽  
H Fujita ◽  
H Suzuki ◽  
...  
Keyword(s):  
Simian Virus 40 ◽  
Soft Agar ◽  
Simian Virus ◽  
T Antigen ◽  
Oncogenic Transformation ◽  
3T3 Cells ◽  
Cellular Mechanisms ◽  
Ras Gene ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

A flat revertant, R1, was isolated from human activated c-Ha-ras-1 (hu-ac-Ha-ras) gene-transformed NIH 3T3 cells (EJ-NIH 3T3) treated with mutagens. R1 contained unchanged transfected hu-ac-Ha-ras DNA and expressed high levels of hu-ac-Ha-ras-specific mRNA and p21 protein. Transfection experiments revealed that NIH 3T3 cells could be transformed by DNA from R1 cells but R1 cells could not be retransformed by Kirsten sarcoma virus, DNA from EJ-NIH 3T3 cells, hu-ac-Ha-ras, v-src, v-mos, simian virus 40 large T antigen, or polyomavirus middle T antigen. Somatic cell hybridization studies showed that R1 was not retransformed by fusion with NIH 3T3 cells and suppressed anchorage independence of EJ-NIH 3T3 and hu-ac-Ha-ras gene-transformed rat W31 cells in soft agar. These results suggest that the reversion and resistance to several oncogenes in R1 is due not to cellular defects in the production of the transformed phenotype but rather to enhancement of cellular mechanisms that suppress oncogenic transformation.

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