Changes in Protein Expression in Two Cholangiocarcinoma Cell Lines Undergoing Formation of Multicellular Tumor Spheroids In Vitro

Mischiati, Carlo; Ura, Blendi; Roncoroni, Leda; Elli, Luca; Cervellati, Carlo; Squerzanti, Monica; Dario, Conte; Doneda, Luisa; Laureto, Patrizia Polverino de; Franceschi, Giorgia De; Calza, Roberta; Barrero, Carlos; Merali, Salim; Ferrari, Carlo; Bergamini, Carlo M.; Agostinelli, Enzo

doi:10.1371/journal.pone.0118906

Cited by 17 publications

(21 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Western blot analysis was performed as previously described (16). An equal amount of protein to that used for 2-DE analysis (30 µg) was separated by 12% SDS-PAGE and then transferred to a nitrocellulose membrane.…”

Section: Trypsin Digestion and Matrix-assisted Laser Desorption/ionizmentioning

confidence: 99%

“…Ingenuity Pathway Analysis (IPA). Changes in protein abundance identified by MS in leiomyoma IF were analyzed by IPA (Qiagen GmbH, Hilden, Germany) as previously described (16). Selected genes were incorporated in canonical pathways and bio-functions, and were used to generate biological networks.…”

Section: Trypsin Digestion and Matrix-assisted Laser Desorption/ionizmentioning

confidence: 99%

See 1 more Smart Citation

Identification of proteins with different abundance associated with cell migration and proliferation in leiomyoma interstitial fluid by proteomics

et al. 2017

Self Cite

View full text Add to dashboard Cite

Abstract. Uterine leiomyoma is the most common female reproductive tract benign tumor. Little is known about protein composition and changes in the leiomyoma interstitial fluid (IF). The present study focused on changes in protein abundance in the IF of leiomyoma. Leiomyoma IFs and adjacent myometrial IFs were obtained and analyzed by two-dimensional electrophoresis (2-DE) coupled with mass spectrometry and western blotting for 2-DE data validation. A total of 25 unique proteins were observed to change significantly (P<0.05). Of these proteins with different abundance, 22 had not been previously identified in leiomyoma IF. In silico analysis predicted that three of these proteins were secreted via classical mechanisms, while 22 were secreted via non-classical mechanisms. Ingenuity Pathway Analysis identified 17 proteins associated with cellular migration and proliferation. Among these, phosphoglycerate mutase 1 had not been previously associated with leiomyoma. The abundance of seven proteins was further validated by western blotting. A comparative proteomic approach identified a number of proteins associated with cellular migration and proliferation, with changes in abundance in IF likely to be involved in tumor development. Further studies will be required to investigate the role of these proteins in leiomyoma IF and their possible association with tumor development and growth.

show abstract

Section: Trypsin Digestion and Matrix-assisted Laser Desorption/ionizmentioning

confidence: 99%

Section: Trypsin Digestion and Matrix-assisted Laser Desorption/ionizmentioning

confidence: 99%

Identification of proteins with different abundance associated with cell migration and proliferation in leiomyoma interstitial fluid by proteomics

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…Western blotting. Western blot analysis was performed as previously described (12). Briefly, an equal amount of protein (30 µg) used for 2-DE analysis was separated by 12% SDS-PAGE and then transferred to a nitrocellulose membrane.…”

Section: Dysregulated Chaperones Associated With Cell Proliferation Amentioning

confidence: 99%

Dysregulated chaperones associated with cell proliferation and negative apoptosis regulation in the uterine leiomyoma

et al. 2018

Self Cite

View full text Add to dashboard Cite

Uterine leiomyomas are benign smooth muscle cell tumors that originate from the myometrium. In this study we focus on dysregulated chaperones associated with cell proliferation and apoptosis. Paired tissue samples of 15 leiomyomas and adjacent myometria were obtained and analyzed by two-dimensional gel electrophoresis (2-DE). Mass spectrometry was used for protein identification and western blotting for 2-DE data validation. The values of 6 chaperones were found to be significantly different in the leiomyoma when compared with the myometrium. A total of 4 proteins were upregulated in the leiomyoma and 2 proteins were downregulated. Calreticulin and 78 kDa glucose-regulated protein were further validated by western blotting because the first is considered a marker of cell proliferation, while the second protects against apoptotic cell death. In addition, we also validated the two downregulated proteins heat shock protein β-1 and heat shock 70 kDa protein 1A. Our study shows the existence of a dysregulation of chaperone proteins associated with leiomyoma development. Functional studies are needed to ascertain the role of these chaperones in the leiomyoma. This may be crucial for the further development of specific inhibitors against the activity of these proteins in order to block the growth of the leiomyoma.

show abstract

“…Western blotting. Western blot analysis was performed as previously described (12). Protein extracts (30 µg) used for 2-DE were separated by 10% SDS-PAGE, and then transferred to a nitrocellulose membrane in a blotting chamber.…”

Section: String 90 Network Analysis and Biological Functionsmentioning

confidence: 99%

Abnormal expression of leiomyoma cytoskeletal proteins involved in cell migration

et al. 2016

Self Cite

View full text Add to dashboard Cite

Uterine leiomyomas are monoclonal tumors. Several factors are involved in the neoplastic transformation of the myometrium. In our study we focused on dysregulated cytoskeletal proteins in the leiomyoma as compared to the myometrium. Paired tissue samples of ten leiomyomas and adjacent myometria were obtained and analyzed by two‑dimensional gel electrophoresis (2-DE). Mass spectrometry was used for protein identification, and western blotting for 2-DE data validation. The values of ten cytoskeletal proteins were found to be significantly different: eight proteins were upregulated in the leiomyoma and two proteins were downregulated. Three of the upregulated proteins (myosin regulatory light polypeptide 9, four and a half LIM domains protein 1 and LIM and SH3 domain protein 1) are involved in cell migration, while downregulated protein transgelin is involved in replicative senescence. Myosin regulatory light polypeptide 9 (MYL9) was further validated by western blotting because it is considered to be a cell migration marker in several cancers and could play a key role in leiomyoma development. Our data demonstrate significant alterations in the expression of cytoskeletal proteins involved in leiomyoma growth. A better understanding of the involvement of cytoskeletal proteins in leiomyoma pathogenesis may contribute to the identification of new therapeutic targets and the development of new pharmacological approaches.

show abstract

Changes in Protein Expression in Two Cholangiocarcinoma Cell Lines Undergoing Formation of Multicellular Tumor Spheroids In Vitro

Cited by 17 publications

References 55 publications

Identification of proteins with different abundance associated with cell migration and proliferation in leiomyoma interstitial fluid by proteomics

Identification of proteins with different abundance associated with cell migration and proliferation in leiomyoma interstitial fluid by proteomics

Dysregulated chaperones associated with cell proliferation and negative apoptosis regulation in the uterine leiomyoma

Abnormal expression of leiomyoma cytoskeletal proteins involved in cell migration

Contact Info

Product

Resources

About