Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Molecular Characterization of ANKRD1 in Rhabdomyosarcoma Cell Lines: Expression, Localization, and Proteasomal Degradation

Emilija Milosevic
ORCID logo ,
Mirjana Novkovic
,
Vittoria Cenni
,
Alberto Bavelloni
ORCID logo ,
Snezana Kojic
* ORCID logo ,
Jovana Jasnic
* ORCID logo
Version 1 : Received: 8 September 2023 / Approved: 8 September 2023 / Online: 11 September 2023 (09:28:07 CEST)

A peer-reviewed article of this Preprint also exists.

Milosevic, E.; Novkovic, M.; Cenni, V.; Bavelloni, A.; Kojic, S.; Jasnic, J. Molecular Characterization of ANKRD1 in Rhabdomyosarcoma Cell Lines: Expression, Localization, and Proteasomal Degradation. Histochemistry and Cell Biology 2024, doi:10.1007/s00418-024-02272-2. Milosevic, E.; Novkovic, M.; Cenni, V.; Bavelloni, A.; Kojic, S.; Jasnic, J. Molecular Characterization of ANKRD1 in Rhabdomyosarcoma Cell Lines: Expression, Localization, and Proteasomal Degradation. Histochemistry and Cell Biology 2024, doi:10.1007/s00418-024-02272-2.

Abstract

Rhabdomyosarcoma (RMS) is the most common soft tissue malignancy in children and adolescents. Respecting the age of the patients and the tumor aggressiveness, investigation of the molecular mechanisms of RMS tumorigenesis is directed toward the identification of novel therapeutic targets. To contribute to a better understanding of the molecular pathology of RMS, we investigated ankyrin repeat domain 1 (ANKRD1), designated as a potential marker for differential diagnostics. In this study, we used three RMS cell lines (SJRH30, RD, and HS-729) to assess its expression profile, intracellular localization, and turnover. They express wild-type ANKRD1, as judged by the sequencing of the open reading frame. Each cell line expressed a different amount of ANKRD1 protein, although the transcript level was similar. According to Western blot analysis, ANKRD1 protein was expressed at detectable levels in the SJRH30 and RD cells (SJRH30>RD), but not in the HS-729, even after immunoprecipitation. Immunocytochemistry revealed nuclear and cytoplasmic localization of ANKRD1 in all examined cell lines. Moreover, the punctate pattern of ANKRD1 staining in the nuclei of RD and HS-729 cells overlapped with coilin, indicating its association with Cajal bodies. We have shown that RMS cells are not able to overexpress ANKRD1 protein, which can be attributed to its proteasomal degradation. The unsuccessful attempt to overexpress ANKRD1 in RMS cells indicates the possibility that its overexpression may be lethal for RMS cells and opens a window for therapeutic targeting.

Keywords

rhabdomyosarcoma; ANKRD1; Cajal bodies; proteasomal degradation

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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