Effect of TTLL12 on tubulin tyrosine nitration as a novel target for screening anticancer drugs<em> in&nbsp;vitro</em>

Abstract

Nitrotyrosine, a structural analogue of tyrosine, is present in cells in pathological conditions and is incorporated into tubulin to form tubulin tyrosine nitration, which disrupts the normal function of microtubules. There is limited research on the functional aspects of tubulin tyrosine nitration in different types of tumor. In the present study, the effect of tubulin tyrosine nitration and tubulin tyrosine ligase like 12 (TTLL12) on the proliferation of SCC-25 cells was investigated. TTLL12-overexpressing cell lines were constructed and used to assess the effect of tubulin tyrosine nitration and TTLL12 on the proliferation of SCC-25 cells via western blotting, immunofluorescent and MTT assays. An TTLL12-stably overexpressing SCC-25 cell line and the enzyme-linked immunosorbent assay were used to establish a novel experiment in vitro for screening anticancer drugs targeting tubulin tyrosine nitration by assessing its sensitivity, specificity and repeatability, and using it to find an effective drug. The results demonstrated that the proliferative rate of the control cells was notably inhibited in the presence of nitrotyrosine compared with that of TTLL12-overexpressing cells. The results of the MTT assay revealed that the proliferation of TTLL12-silenced cells was significantly inhibited compared with that of the control group. The sensitivity, specificity and repeatability of the experiment were positive. It was found that nocodazole could have better anticancer effect than paclitaxel. Taken together, the results of the present study suggest that TTLL12 enhances SCC-25 cell survival in the presence of nitrotyrosine by disrupting nitration of the tyrosine residues of tubulin, and tubulin tyrosine nitration may be developed for the basic research of anticancer drugs.

Inhibition of Nitrosative Stress Attenuates Myocardial Injury and Improves Outcomes after Cardiac Arrest and Resuscitation

“…ONOO À can nitrate protein tyrosine to produce 3nitrotyrosine (3-NT), which is a specific biomarker of nitrosative stress. The nitration modification of proteins can regulate protein function by affecting the biological activity, expression, subcellular localization, and intermolecular interactions of the protein and then affects various pathophysiological processes of the cell (16,17). These results suggest that suppression of nitrosative stress is a very important way to protect the myocardium from ischemia/reperfusion injury.…”

TTLL12 has a potential oncogenic activity, suppression of ligation of nitrotyrosine to the C-terminus of detyrosinated α-tubulin, that can be overcome by molecules identified by screening a compound library

“…The data is this manuscript are original and were first reported in a thesis in 2009 [48]. They have since been replicated by a previous member of the laboratory who was aware of these results and worked on other TTLL12 related project in our laboratory [59,60]. BW is an Emeritus Research Director in the CNRS, affiliated to the IGBMC.…”

Novel inhibitors of TTLL12’s oncogenic potential that overcome suppression of ligation of nitrotyrosine to the C-terminus of detyrosinated α-tubulin

“…The data is this manuscript are original and were first reported in a thesis in 2009 (47). They have since been replicated by a previous member of the laboratory who was aware of these results and worked on other TTLL12 related project in our laboratory (58,59). BW is an Emeritus Research Director in the CNRS, affiliated to the IGBMC.…”