The transformics assay: first steps for the development of an integrated approach to investigate the malignant cell transformation in vitro

Abstract Data from epidemiological studies suggest that consumption of red and processed meat is a factor contributing to colorectal carcinogenesis. Red meat contains high amounts of heme, which in turn can be converted to its nitrosylated form, NO-heme, when adding nitrite-containing curing salt to meat. NO-heme might contribute to colorectal cancer formation by causing gene mutations and could thereby be responsible for the association of (processed) red meat consumption with intestinal cancer. Up to now, neither in vitro nor in vivo studies characterizing the mutagenic and cell transforming potential of NO-heme have been published due to the fact that the pure compound is not readily available. Therefore, in the present study, an already existing synthesis protocol was modified to yield, for the first time, purified NO-heme. Thereafter, newly synthesized NO-heme was chemically characterized and used in various in vitro approaches at dietary concentrations to determine whether it can lead to DNA damage and malignant cell transformation. While NO-heme led to a significant dose-dependent increase in the number of DNA strand breaks in the comet assay and was mutagenic in the HPRT assay, this compound tested negative in the Ames test and failed to induce malignant cell transformation in the BALB/c 3T3 cell transformation assay. Interestingly, the non-nitrosylated heme control showed similar effects, but was additionally able to induce malignant transformation in BALB/c 3T3 murine fibroblasts. Taken together, these results suggest that it is the heme molecule rather than the NO moiety which is involved in driving red meat-associated carcinogenesis.

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Long-term exposures to low doses of silver nanoparticles enhanced in vitro malignant cell transformation in non-tumorigenic BEAS-2B cells

Toxicology in Vitro ◽

10.1016/j.tiv.2016.09.003 ◽

2016 ◽

Vol 37 ◽

pp. 41-49 ◽

Cited By ~ 28

Author(s):

Wun Hak Choo ◽

Cho Hee Park ◽

Shi Eun Jung ◽

Byeonghak Moon ◽

Huiyeon Ahn ◽

...

Keyword(s):

Silver Nanoparticles ◽

Cell Transformation ◽

Malignant Cell ◽

Low Doses ◽

Malignant Cell Transformation

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Induction of malignant cell transformation in vitro by extract and fractions of airborne particulate matter

Journal of Aerosol Science ◽

10.1016/0021-8502(86)90104-7 ◽

1986 ◽

Vol 17 (3) ◽

pp. 356-360 ◽

Cited By ~ 13

Author(s):

N.H. Seemayer ◽

N. Manojlovic ◽

R. Tomingas

Keyword(s):

Particulate Matter ◽

Cell Transformation ◽

Airborne Particulate Matter ◽

Malignant Cell ◽

Airborne Particulate ◽

Malignant Cell Transformation

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Chondroitin Sulphate Proteoglycan 4 (NG2/CSPG4) Localization in Low- and High-Grade Gliomas

Cells ◽

10.3390/cells9061538 ◽

2020 ◽

Vol 9 (6) ◽

pp. 1538 ◽

Cited By ~ 1

Author(s):

Marta Mellai ◽

Laura Annovazzi ◽

Ilaria Bisogno ◽

Cristiano Corona ◽

Paola Crociara ◽

...

Keyword(s):

Cell Transformation ◽

Chondroitin Sulphate ◽

Molecular Genetic ◽

Malignant Gliomas ◽

Malignant Cell ◽

Wild Type ◽

Sulphate Proteoglycan ◽

Chondroitin Sulphate Proteoglycan ◽

Proteoglycan 4

Background: Neuron glial antigen 2 or chondroitin sulphate proteoglycan 4 (NG2/CSPG4) is expressed by immature precursors/progenitor cells and is possibly involved in malignant cell transformation. The aim of this study was to investigate its role on the progression and survival of sixty-one adult gliomas and nine glioblastoma (GB)-derived cell lines. Methods: NG2/CSPG4 protein expression was assessed by immunohistochemistry and immunofluorescence. Genetic and epigenetic alterations were detected by molecular genetic techniques. Results: NG2/CSPG4 was frequently expressed in IDH-mutant/1p19q-codel oligodendrogliomas (59.1%) and IDH-wild type GBs (40%) and rarely expressed in IDH-mutant or IDH-wild type astrocytomas (14.3%). Besides tumor cells, NG2/CSPG4 immunoreactivity was found in the cytoplasm and/or cell membranes of reactive astrocytes and vascular pericytes/endothelial cells. In GB-derived neurospheres, it was variably detected according to the number of passages of the in vitro culture. In GB-derived adherent cells, a diffuse positivity was found in most cells. NG2/CSPG4 expression was significantly associated with EGFR gene amplification (p = 0.0005) and poor prognosis (p = 0.016) in astrocytic tumors. Conclusion: The immunoreactivity of NG2/CSPG4 provides information on the timing of the neoplastic transformation and could have prognostic and therapeutic relevance as a promising tumor-associated antigen for antibody-based immunotherapy in patients with malignant gliomas.

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Effect and mechanism of YB-1 knockdown on glioma cell growth, migration, and apoptosis

Acta Biochimica et Biophysica Sinica ◽

10.1093/abbs/gmz161 ◽

2020 ◽

Vol 52 (2) ◽

pp. 168-179 ◽

Cited By ~ 1

Author(s):

Huilin Gong ◽

Shan Gao ◽

Chenghuan Yu ◽

Meihe Li ◽

Ping Liu ◽

...

Keyword(s):

Cell Cycle ◽

Cell Proliferation ◽

Cell Growth ◽

Glioma Cell ◽

Cell Transformation ◽

Malignant Cell ◽

Protein Levels ◽

Glioma Progression

Abstract Y-box binding protein 1 (YB-1) is manifested as its involvement in cell proliferation and differentiation and malignant cell transformation. Overexpression of YB-1 is associated with glioma progression and patient survival. The aim of this study is to investigate the influence of YB-1 knockdown on glioma cell progression and reveal the mechanisms of YB-1 knockdown on glioma cell growth, migration, and apoptosis. It was found that the knockdown of YB-1 decreased the mRNA and protein levels of YB-1 in U251 glioma cells. The knockdown of YB-1 significantly inhibited cell proliferation, colony formation, and migration in vitro and tumor growth in vivo. Proteome and phosphoproteome data revealed that YB-1 is involved in glioma progression through regulating the expression and phosphorylation of major proteins involved in cell cycle, adhesion, and apoptosis. The main regulated proteins included CCNB1, CCNDBP1, CDK2, CDK3, ADGRG1, CDH-2, MMP14, AIFM1, HO-1, and BAX. Furthermore, it was also found that YB-1 knockdown is associated with the hypo-phosphorylation of ErbB, mTOR, HIF-1, cGMP-PKG, and insulin signaling pathways, and proteoglycans in cancer. Our findings indicated that YB-1 plays a key role in glioma progression in multiple ways, including regulating the expression and phosphorylation of major proteins associated with cell cycle, adhesion, and apoptosis.

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Inhibition of the aurora kinases suppresses in vitro NT2-D1 cell growth and tumorigenicity

Journal of Endocrinology ◽

10.1677/joe-09-0257 ◽

2009 ◽

Vol 204 (2) ◽

pp. 135-142 ◽

Cited By ~ 8

Author(s):

Salvatore Ulisse ◽

Yannick Arlot-Bonnemains ◽

Enke Baldini ◽

Stefania Morrone ◽

Silvia Carocci ◽

...

Keyword(s):

Cell Cycle ◽

Cell Proliferation ◽

Cell Transformation ◽

Germ Cell Tumors ◽

Aurora Kinase ◽

Malignant Cell ◽

Dependent Manner ◽

Testicular Germ Cell ◽

Aurora Kinases

The aurora kinase family members, Aurora-A, -B, and -C (listed as AURKA, AURKB and AURKC respectively in the HUGO Database), are serine/threonine kinases involved in the regulation of chromosome segregation and cytokinesis, and alterations in their expression are associated with malignant cell transformation and genomic instability. Deregulation of the expression of the aurora kinases has been shown to occur also in testicular germ cell tumors (TGCTs) identifying them as putative anticancer therapeutic targets. We here evaluated the in vitro effects of MK-0457, an aurora kinases inhibitor, on cell proliferation, cell cycle, ploidy, apoptosis, and tumorigenicity on the TGCT-derived cell line NT2-D1. Treatment with MK-0457 inhibited cell proliferation in a time- and dose-dependent manner, with IC50=17.2±3.3 nM. MK-0457 did not affect the expression of the three aurora kinases, but prevented their ability to phosphorylate substrates relevant to the mitotic progression. Time-lapse experiments demonstrated that MK-0457-treated cells entered mitosis but were unable to complete it, presenting after short time the typical features of apoptotic cells. Cytofluorimetric analysis confirmed that the treatment with MK-0457 for 6 h induced NT2-D1 cells accumulation in the G2/M phase of the cell cycle and the subsequent appearance of sub-G0 nuclei. The latter result was further supported by the detection of caspase-3 activation following 24-h treatment with the inhibitor. Finally, MK-0457 prevented the capability of the NT2-D1 cells to form colonies in soft agar. In conclusion, the above findings demonstrate that inhibition of aurora kinase activity is effective in reducing in vitro growth and tumorigenicity of NT2-D1 cells, and indicate its potential therapeutic value for TGCT treatment.

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