ShujaaEdin Hareth Y, Al-Haj Nagi A, Rasedee Abdullah, Alitheen Noorjahan Banu, Kadir Arifah Abdul, How Chee Wun, Rahman Heshu Sulaiman, Abdullah Al-Shwyeh Hussah
Institute of Bioscience, Universiti Putra Malaysia, Malaysia.
Faculty of Medicine and Health Sciences, Sana'a University, Yemen.
Saudi J Biol Sci. 2021 Apr;28(4):2549-2557. doi: 10.1016/j.sjbs.2021.01.059. Epub 2021 Feb 11.
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.
重组人促红细胞生成素（rHuEPO）是一种促红细胞生成激素，目前在癌症贫血治疗中与化疗药物联合使用。rHuEPO对三维（3D）培养的癌细胞的影响尚不清楚。本研究的目的是确定rHuEPO对二维（2D）和3D细胞培养的MCF-7乳腺癌细胞活力的影响。用0、0.1、10、100或200 IU/mL的rHuEPO处理二维培养的单层MCF-7细胞和通过超低粘附微孔板技术生成的三维培养的MCF-7细胞24、48或72小时。使用（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四氮唑（MTT）法、中性红保留时间（NRRT）、台盼蓝排斥试验（TBE）、DNA片段化、吖啶橙/碘化丙啶染色（AO/PI）试验来测定rHuEPO对MCF-7细胞活力和增殖的影响。对用于三维培养的MCF-7细胞也进行了半胱天冬酶试验和流式细胞术细胞周期分析。rHuEPO对降低三维培养的MCF-7细胞活力的作用似乎比二维培养的更大。rHuEPO以剂量和时间依赖性方式显著（p<0.05）降低了三维MCF-7细胞的活力并下调了其半胱天冬酶活性。细胞周期分析表明，rHuEPO使MCF-7细胞进入亚G0/G1期。因此，该研究表明rHuEPO对三维培养的MCF-7乳腺癌细胞具有细胞生长抑制作用。
