TY - JOUR
T1 - Human breast tumor derived endothelial cells exhibit distinct biological properties
AU - Hegde, Mangala
AU - Bhat, Sharath Mohan
AU - Guruprasad, Kanive Parashiva
AU - Moka, Rajasekhar
AU - Ramachandra, Lingadakai
AU - Satyamoorthy, Kapaettu
AU - Joshi, Manjunath B.
N1 - Funding Information:
Authors thank TIFAC‐Core, Government of India and Manipal Academy for Higher Education, Manipal for infrastructure. We gratefully acknowledge the gift of sorafenib from Cyno Pharmaceuticals, New Delhi. We thank Ms. Jyothi, Ms. Soumya, and Mr. Ganesh for technical assistance. The project was funded by Cancer biology‐Pilot studies grant from Department of Biotechnology, Government of India (6242‐P81/RGCB/PMD/DBT/MNBJ/2015). Mangala Hegde was financially supported by Dr. TMA Pai Fellowship, MAHE, Manipal and Senior Research Fellowship, CSIR, Government of India. Sharath M Bhat is financially supported by Dr. TMA Pai Fellowship, MAHE, Manipal.
Funding Information:
Authors thank TIFAC-Core, Government of India and Manipal Academy for Higher Education, Manipal for infrastructure. We gratefully acknowledge the gift of sorafenib from Cyno Pharmaceuticals, New Delhi. We thank Ms. Jyothi, Ms. Soumya, and Mr. Ganesh for technical assistance. The project was funded by Cancer biology-Pilot studies grant from Department of Biotechnology, Government of India (6242-P81/RGCB/PMD/DBT/MNBJ/2015). Mangala Hegde was financially supported by Dr. TMA Pai Fellowship, MAHE, Manipal and Senior Research Fellowship, CSIR, Government of India. Sharath M Bhat is financially supported by Dr. TMA Pai Fellowship, MAHE, Manipal.
Publisher Copyright:
© 2021 Société Française des Microscopies and Société de Biologie Cellulaire de France. Published by John Wiley & Sons Ltd
PY - 2022/2
Y1 - 2022/2
N2 - Background Information: Excessive angiogenesis characterized by leaky, tortuous, and chaotic vasculature is one of the hallmarks of cancers and is significantly correlated to poor prognosis. Disorganized angiogenesis leads to poor perfusion of anti-cancer drugs and limits access to immune cells. Hence, impeding angiogenesis is one of the attractive therapeutic targets to inhibit progression and metastasis in several solid tumors including breast. Results: We have developed a robust and reproducible method for isolating and ex vivo culture of endothelial cells (EC) derived from non-malignant (Endo-N) and malignant (Endo-T) part from clinically characterized human breast tumors. RT-PCR and immunoblotting analysis indicated that these cells exhibited expression of endothelial specific genes such as PECAM-1 (CD31), Endoglin (CD105), eNOS, VE-cadherin, VCAM1, and MCAM. Vasculogenic mimicry and contamination of progenitor EC recruited in tumors was ruled out by absence of CD133 expression and normal karyotype. Both the cell types showed stable expression of CD31 and CD105 up to seven passages. Furthermore, compared to Endo-N cells, Endo-T cells showed (a) constitutively increased proliferation marked by nearly 36% of cells in mitotic phase, (b) requirement of glutamine for cell survival, (c) pro-migratory phenotype, (d) produced increased number of sprouts in 3D cultures, and (e) resistance to sorafenib. Conclusion: Tumor derived EC showed distinct biological properties compared to normal breast EC. Significance: Our method for isolating endothelial cell types from human breast tumors may be explored to (a) understand cellular and molecular mechanisms, (b) screen anti-angiogenic molecules, and (c) formulate organoid cultures to develop personalized medicine facilitating better clinical management of breast cancers.
AB - Background Information: Excessive angiogenesis characterized by leaky, tortuous, and chaotic vasculature is one of the hallmarks of cancers and is significantly correlated to poor prognosis. Disorganized angiogenesis leads to poor perfusion of anti-cancer drugs and limits access to immune cells. Hence, impeding angiogenesis is one of the attractive therapeutic targets to inhibit progression and metastasis in several solid tumors including breast. Results: We have developed a robust and reproducible method for isolating and ex vivo culture of endothelial cells (EC) derived from non-malignant (Endo-N) and malignant (Endo-T) part from clinically characterized human breast tumors. RT-PCR and immunoblotting analysis indicated that these cells exhibited expression of endothelial specific genes such as PECAM-1 (CD31), Endoglin (CD105), eNOS, VE-cadherin, VCAM1, and MCAM. Vasculogenic mimicry and contamination of progenitor EC recruited in tumors was ruled out by absence of CD133 expression and normal karyotype. Both the cell types showed stable expression of CD31 and CD105 up to seven passages. Furthermore, compared to Endo-N cells, Endo-T cells showed (a) constitutively increased proliferation marked by nearly 36% of cells in mitotic phase, (b) requirement of glutamine for cell survival, (c) pro-migratory phenotype, (d) produced increased number of sprouts in 3D cultures, and (e) resistance to sorafenib. Conclusion: Tumor derived EC showed distinct biological properties compared to normal breast EC. Significance: Our method for isolating endothelial cell types from human breast tumors may be explored to (a) understand cellular and molecular mechanisms, (b) screen anti-angiogenic molecules, and (c) formulate organoid cultures to develop personalized medicine facilitating better clinical management of breast cancers.
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U2 - 10.1111/boc.202100015
DO - 10.1111/boc.202100015
M3 - Article
AN - SCOPUS:85119659622
SN - 0248-4900
VL - 114
SP - 73
EP - 85
JO - Biology of the Cell
JF - Biology of the Cell
IS - 2
ER -
