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Recombinant Human erythropoietin reduces viability of MCF-7 breast cancer cells from 3D culture without caspase activation
Authors:Hareth Y ShujaaEdin  Nagi A AL-Haj  Abdullah Rasedee  Noorjahan Banu Alitheen  Arifah Abdul Kadir  Chee Wun How  Heshu Sulaiman Rahman  Al-Shwyeh Hussah Abdullah
Institution:1. Institute of Bioscience, Universiti Putra Malaysia, Malaysia;2. Faculty of Medicine and Health Sciences, Sana’a University, Yemen;3. Faculty of Veterinary Medicine, Universiti Putra Malaysia, Malaysia;4. Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, Malaysia;5. School of Pharmacy, Monash University Malaysia, Bandar Sunway, Selangor, Malaysia;6. College of Medicine, University of Sulaimani, Kurdistan Region, Iraq;7. College of Science, Imam Abdul Rahman bin Faisal University, Saudi Arabia
Abstract:Recombinant human erythropoietin (rHuEPO) is the erythropoiesis-stimulating hormone that is being used concurrently with chemotherapeutic drugs in the treatment of anemia of cancer. The effect of rHuEPO on cancer cells in 3-dimensional (3D) cultures is not known. The objective of the study was to determine the effect of rHuEPO on the viability of MCF-7 breast cancer cells from 2-dimensional (2D) and 3D cell cultures. The monolayer MCF-7 cells from 2D culture and MCF-7 cell from 3D culture generated by ultra-low adhesive microplate technique, were treated with 0, 0.1, 10, 100 or 200 IU/mL rHuEPO for 24, 48 or 72 h. The effects of rHuEPO on MCF-7 cell viability and proliferation were determined using the (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay (MTT), neutral red retention time (NRRT), trypan blue exclusion assay (TBE), DNA fragmentation, acridine orange/propidium iodide staining (AO/PI) assays. The MCF-7 cells for 3D culture were also subjected to caspase assays and cell cycle analysis using flow cytometry. rHuEPO appeared to have greater effect at lowering the viability of MCF-7 cells from 3D than 2D cultures.rHuEPO significantly (p < 0.05) decreased viability and down-regulated the caspase activities of 3D MCF-7 cells in dose- and time-dependent manner. The cell cycle analysis showed that rHuEPO caused MCF-7 cells to enter the subG0/G1 phase. Thus, the study suggests that rHuEPO has a cytostatic effect on the MCF-7 breast cancer cells from 3D culture.
Keywords:Recombinant human erythropoietin  MCF-7 cell  2-dimensional MCF-7 cell culture  3-dimensional MCF-7 cell culture  rHuEPO"}  {"#name":"keyword"  "$":{"id":"k0030"}  "$$":[{"#name":"text"  "_":"recombinant human erythropoietin  MTT"}  {"#name":"keyword"  "$":{"id":"k0040"}  "$$":[{"#name":"text"  "_":"(4  5-dimethylthiazol-2-yl)-2  5-diphenyl tetrazolium bromide assay  NRR"}  {"#name":"keyword"  "$":{"id":"k0050"}  "$$":[{"#name":"text"  "_":"neutral red retention  TBE"}  {"#name":"keyword"  "$":{"id":"k0060"}  "$$":[{"#name":"text"  "_":"trypan blue exclusion  AO/PI"}  {"#name":"keyword"  "$":{"id":"k0070"}  "$$":[{"#name":"text"  "_":"acridine orange/propidium iodide staining  ULAT"}  {"#name":"keyword"  "$":{"id":"k0080"}  "$$":[{"#name":"text"  "_":"ultra-low adhesion technique  FBS"}  {"#name":"keyword"  "$":{"id":"k0090"}  "$$":[{"#name":"text"  "_":"fetal bovine serum
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