Nanoencapsulation of Hirudo medicinalis proteins in liposomes as a nanocarrier for inhibiting angiogenesis through targeting VEGFA in the Breast cancer cell line (MCF-7)

Nanoencapsulation of Hirudo medicinalis proteins in liposomes as a nanocarrier for inhibiting angiogenesis through targeting VEGFA in the Breast cancer cell line (MCF-7) Amir Shakouri Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Department of Molecular Medicine, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran http://orcid.org/0000-0002-5893-2340 Houman Kahroba Department of Molecular Medicine, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran Hamed Hamishekar Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Jalal Abdolalizadeh Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Paramedical Faculty, Tabriz University of Medical Sciences, Tabriz, Iran http://orcid.org/0000-0002-8283-5057

Introduction: Breast cancer is the most serious cause of women’s death throughout the world. Using nanocarrier vehicles to the exact site of cancer upgrades the therapeutic efficiency of the drugs. Capsulation of active proteins in the vesicular liposomes’ hydrophilic core is essential to develop a therapeutic protein carrier system. We aimed to encapsulate the medicinal leech saliva extract (LSE) and assess the inhibition of angiogenesis of breast cancer cells by targeting vascular endothelial growth factor A (VEGFA). Methods: In this research, enhanced formulation of liposomal protein was determined by zeta potential analysis, droplet size, drug release assay, and transmission electron microscopy (TEM). Furthermore, a cytotoxicity assay of liposomal LSE was performed to determine the cytotoxic activity of components. For assessing the expression of VEGFA, P53, and hypoxia-inducible factor subunit alpha (HIF1a) genes, Real-Time PCR was applied. Results: Nano liposome was chosen as an enhanced formulation due to its much smaller size (46.23 nm). Liposomal LSE had more practical actions on the MCF-7 cells. As noticed by DAPI staining, apoptosis was extensively greater in treated MCF-7 cells. Wound healing assay demonstrated that MCF-7 cells could not sustain growth at the presence of liposomal LSE and expression of the VEGFA gene was declined in treated cells. Downregulation of VEGFA was evaluated with western blotting technique. Conclusion: It can be concluded that our investigation of the tests confirmed the fact that nano liposomal LSE is a novel promising formulation for anticancer drugs and can significantly improve the penetration of protein drugs to cancer cells.
08 09 2021 08 09 2021 115 126 1 10.34172/crossmark_policy bi.tbzmed.ac.ir false Tabriz University of Medical Sciences Tabriz University of Medical Sciences 2020-07-20 2020-12-09 2021-08-09 10.34172/bi.2021.39 20220328130832 https://bi.tbzmed.ac.ir/Article/bi-23253 https://bi.tbzmed.ac.ir/PDF/bi-12-115.pdf https://bi.tbzmed.ac.ir/PDF/bi-12-115.pdf https://bi.tbzmed.ac.ir/PDF/bi-12-115.pdf https://bi.tbzmed.ac.ir/PDF/bi-12-115.pdf https://bi.tbzmed.ac.ir/PDF/bi-12-115.pdf 10.3390/molecules23040907 Nano Research Nanoscale chemical imaging of individual chemotherapeutic cytarabine-loaded liposomal nanocarriers Wieland K 12 197 2019 10.1007/s12274-018-2202-x 10.1016/j.jconrel.2019.12.023 10.1016/j.ijpharm.2020.119269 10.1002/jcp.25961 10.2217/nnm-2016-0393 10.1016/j.jconrel.2018.02.040 10.2147/IJN.S203330 10.1016/j.jconrel.2019.05.018 10.1016/j.ejps.2017.03.003 10.1016/j.ejpb.2017.02.001 10.1016/j.ejpb.2017.06.016 Chen MJ, Hui H-W, Lee T, Kurtulik P, Surapaneni S. Nanosuspension of a poorly soluble drug via microfluidization process. Google Patents; 2017. 10.1021/acs.molpharmaceut.7b00051 10.1002/jcp.27121 10.1016/B978-1-4831-9937-5.50008-9 Bangham AD. Physical Structure and Behavior of Lipids and Lipid Enzymes. In: Paoletti R, D Kritchevsky, editors. Advances in Lipid Research. Elsevier; 1963. p. 65-104. 10.1016/j.xphs.2017.05.022 10.1186/1475-2859-3-11 10.1002/jcp.25859 10.1016/j.ultsonch.2019.03.018 10.1016/j.bioorg.2017.02.011 10.5681/bi.2014.011 10.4103/0022-3859.74297 Altern Med Rev Bugs as drugs, part two: worms, leeches, scorpions, snails, ticks, centipedes, and spiders Cherniack EP 16 50 2011 10.34172/apb.2021.038 10.1016/j.jaim.2018.09.003 Pharm Anal Acta S Anticancer effects of medical Malaysian leech saliva extract (LSE) Merzouk A 15 2 2012 10.1158/1538-7445.AM2017-107 Ammar A, Guns E, Kucuk O, Abdualkader A, Alaama M, Uddin AH, et al. Mechanism of anticancer activity of BPS-001 (lyophilized leech saliva extract). AACR; 2017. 10.1016/j.bioorg.2019.01.019 J Immunol Recombinant human hyaluronidase PH20 does not stimulate an acute inflammatory response and inhibits lipopolysaccharide-induced neutrophil recruitment in the air pouch model of inflammation Huang Z 192 5285 2014 10.4049/jimmunol.1303060 10.1158/0008-5472.CAN-09-4687 Huang Z, Zhao C, Radi A. Characterization of hyaluronan, hyaluronidase PH20, and HA synthase HAS2 in inflammation and cancer. Inflammation and Cell Signaling 2014; 1: e306.. Cancers Therapeutic targeting of hyaluronan in the tumor stroma Kultti A 4 873 2012 10.3390/cancers4030873 10.1021/cr050247k 10.1016/j.ctcp.2017.12.001 IIUM Engineering Journal Isolation and analytical characterization of local Malaysian leech saliva Alaama M 12 51 2011 10.1016/j.chemphyslip.2010.01.005 10.1002/jcp.27622 10.1016/j.colsurfb.2019.04.035 10.1002/jcb.29651 10.1002/jcp.27027 10.1039/c6nr03940e 10.1016/j.ultsonch.2017.08.014 10.1016/j.ultsonch.2018.06.003 10.1080/1061186X.2019.1579819 10.1002/adhm.201701257 10.1080/1061186X.2017.1365873 10.1186/s12934-019-1103-3 10.1080/1061186X.2019.1693578 10.1080/1061186X.2019.1656725 10.1002/adhm.201900564 Pharmaceutics Moyá ML, López-López M, Lebrón JA, Ostos FJ, Pérez D, Camacho V, et alPreparation and characterization of new liposomesBactericidal activity of cefepime encapsulated into cationic liposomes Moyá ML 11 69 2019 10.3390/pharmaceutics11020069 10.1063/1.4962992 10.1016/j.colsurfb.2017.06.022 10.1016/j.ejps.2020.105213 10.1016/j.jconrel.2019.03.015 10.1016/j.ijpharm.2020.119191