PB0781 - Prelatent and Native Antithrombin Effect on Cell Cycle and Proliferation of Glioblastoma Multiforme Cells

Monday, July 11, 2022

6:30 PM – 7:30 PM

Location: Exhibition

Presentation Format: In-Person

Co-Author(s)

JP

Julia Peñas-Martínez

PhD student
Department of Hematology and Medical Oncology, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, CIBERER, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain
Murcia, Murcia, Spain
David Zaragoza Huesca, PhD student

PhD student
Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla
Murcia, Murcia, Spain
EN

Esther Navarro Mazano

PhD Student
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Arrixaca, CIBERER, Murcia.
Murcia, Murcia, Spain
EC

Ernesto José Cuenca Zamora

PhD Student
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Arrixaca, CIBERER, Murcia.
Murcia, Murcia, Spain
SE

Salvador Espín

Technician
Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla
Murcia, Murcia, Spain
VV

Vicente Vicente

Researcher and Medical Doctor
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla, CIBERER, Murcia, Spain
Murcia, Murcia, Spain
AC

Alberto Carmona Bayonas

Medical Doctor
Department of Hematology and Medical Oncology, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, CIBERER, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain.
Murcia, Murcia, Spain

Presenting Author(s)

Irene Martínez Martínez, PhD

Principal Investigator
Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla
Murcia, Murcia, Spain

Background: Besides antithrombin’s (AT) key role on hemostasis, two AT’s conformations, native and prelatent, have the anti-tumour functions. We have recently shown that prelatent AT reduces migration and invasion of glioblastoma multiforme cells U-87 MG (U87) and inhibits the expression of VEGFA, showing a potential anti-angiogenic effect. Prelatent AT also reduces pSTAT3 (Tyr705) and pERK1/2 (Thr202/Tyr204) phosphorilation, and STAT3 levels, which have been associated with less resistance to treatment.

Aims: To investigate the effect of prelatent and native AT on glioblastoma multiforme cells.

Methods: Native and prelatent AT were purified from healthy donor’s plasma. A transcriptome analysis was carried out using Human Clariom D chip. The results were analyzed with Partek Genomics Suite software. Differentially expressed genes had >1.5 fold-change. U87 and U-251 MG (U251) cells were treated with native or prelatent AT (2.16 µM) or phosphate-buffered saline for 12 hours before each experiment. Cell cycle and proliferation were assessed by measuring 7-AAD and BrdU incorporation by flow cytometry. Differences in proteins expression were validated by western blot.

Results: After U87 treatment with prelatent AT, 2467 transcripts were overexpressed and 6738 underexpressed. Remarkably, expression of cyclins and regulatory proteins involved in cell cycle were significantly inhibited. We observed ~10% more events in G0-1 phase, a halfway inhibition of S phase, and a 40.63% cell proliferation reduction after U87 treatment with native AT. However, we did not observe significant results in U251 (Figure 1). Western blot analysis validated the inhibition of the expression of CDK4 after U87 treatment with prelatente AT, but not with the native form (p=0.020) (Figure 2).

Conclusion(s): Native and prelatent AT has versatile anti-tumor properties on U87 cells. In particular, we have demonstrated that AT is able to induces cell cycle arrest, but not inhibiting cell cycle cyclins. These results support the potential therapeutic role of AT in glioblastoma multiforme.