TY - JOUR
T1 - Splice‐variant knock‐out of tgfβ receptors perturbates the proteome of ovarian carcinoma cells
AU - Catane, Liora Jacobs
AU - Moshel, Ofra
AU - Smith, Yoav
AU - Davidson, Ben
AU - Reich, Reuven
N1 - Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/12/1
Y1 - 2021/12/1
N2 - The aim of this study was to analyze the biological role of different transforming growth factor‐β (TGFβ) receptor splice variants in ovarian carcinoma (OC). Specific receptor variant knock-outs (KO) were prepared using the CRISPR/Cas9 genome editing system in two OC cell lines, TβRI variant 1 (TβRІv1) KO in ES‐2 cells and TβRII variant 1 (TβRІIv1) KO in OVCAR‐8 cells. Control and KO cells were compared by proteomic analysis, functional tests, analysis of epithelial–mesenchymal transition (EMT) drivers, and Western blot of signaling proteins. Proteomic analysis revealed significant changes in protein pathways in the KO cells. TβRIv1 KO resulted in a significant reduction in both cellular motility and invasion, while TβRIIv1 KO significantly reduced cellular motility and increased Reactive Oxygen Species (ROS) production. Both receptor variant KOs reduced MET protein levels. Of the EMT drivers, a significant decrease in TWIST protein expression, and increase in SNAIL protein and MALAT1 mRNA levels were observed in the TβRІIv1 KO compared to control. A significant decrease in JNK1 and JNK2 activation was found in the TβRІv1 KO compared to control cells. These findings provide new insight regarding the biological role of the TGFβ receptor variants in the biology and potentially the progression of OC.
AB - The aim of this study was to analyze the biological role of different transforming growth factor‐β (TGFβ) receptor splice variants in ovarian carcinoma (OC). Specific receptor variant knock-outs (KO) were prepared using the CRISPR/Cas9 genome editing system in two OC cell lines, TβRI variant 1 (TβRІv1) KO in ES‐2 cells and TβRII variant 1 (TβRІIv1) KO in OVCAR‐8 cells. Control and KO cells were compared by proteomic analysis, functional tests, analysis of epithelial–mesenchymal transition (EMT) drivers, and Western blot of signaling proteins. Proteomic analysis revealed significant changes in protein pathways in the KO cells. TβRIv1 KO resulted in a significant reduction in both cellular motility and invasion, while TβRIIv1 KO significantly reduced cellular motility and increased Reactive Oxygen Species (ROS) production. Both receptor variant KOs reduced MET protein levels. Of the EMT drivers, a significant decrease in TWIST protein expression, and increase in SNAIL protein and MALAT1 mRNA levels were observed in the TβRІIv1 KO compared to control. A significant decrease in JNK1 and JNK2 activation was found in the TβRІv1 KO compared to control cells. These findings provide new insight regarding the biological role of the TGFβ receptor variants in the biology and potentially the progression of OC.
KW - CRISPR/Cas9 KO’s
KW - Ovarian carcinoma
KW - Proteomics
KW - Receptor splice variants
KW - TGFβ
KW - Transforming growth factor‐β
UR - http://www.scopus.com/inward/record.url?scp=85119672900&partnerID=8YFLogxK
U2 - 10.3390/ijms222312647
DO - 10.3390/ijms222312647
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C2 - 34884451
AN - SCOPUS:85119672900
SN - 1661-6596
VL - 22
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
IS - 23
M1 - 12647
ER -