Shams Azar, Shabani Ronak, Najafi Mohammad, Karimi Mahdi, Pirhajati Vahid, Asghari Jafarabadi Mohammad, Asgari Hamid Reza, Maki Chad B, Razavi Seyed Mohsen, Koruji Morteza
Stem Cell and Regenerative Medicine Research Center, Iran University of Medical Sciences, Tehran, Iran.
Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
Cell J. 2021 Oct;23(5):544-551. doi: 10.22074/cellj.2021.7606. Epub 2021 Oct 30.
In cancer treatments, smart gene delivery via nanoparticles (NPs) can be targeted for cancer cells, while concurrently minimizing damage to healthy cells. This study assessed the efficiency of poly lactic-co-glycolic acid (PLGA)-miR 143/206 transfection on apoptosis in mouse leukemia cancer cells (El4) and spermatogonial stem cells (SSCs).
In this experimental study, neonatal mouse spermatogonia cells and EL4 cancer cell lines were used. MicroRNA-PLGA NPs were prepared, characterized, and targeted with folate. Several doses were evaluated to obtain a suitable miR dose that can induce appropriate apoptosis in EL4 cells, while not harming SSCs. Cells were treated separately at 3 doses of each miR (for miR 143, doses of 25, 50 and 75 nmol and for miR 206, doses of 50, 100 and 150 nmol). The experiments were performed at 24, 48 and 72 hours. Viability and apoptosis were investigated by MTT and Annexin Kits.
Based on MTT assay results, the optimal dose of miR 143 was 75 nmol (59.87 ± 2.85 % SSC and 35.3 ± 0.78 % EL4) (P≤0.05), and for miR 206, the optimal dose was 150 nmol (54.82 ± 6.7 % SSC and 33.92 ± 3.01% EL4) (P≤0.05). The optimal time was 48 hours. At these doses, the survival rate of the EL4 cells was below the half maximal inhibitory concentration (IC) and SSC survival was above 50%. Annexin V staining also confirmed the selected doses (for miR 143 total apoptosis was 6.62% ± 1.8 SSC and 37.4% ± 4.2 EL4 (P≤0.05), and miR 206 was (10.98% ± 1.5 SSC and 36.4% ± 3.7 EL4, P≤0.05).
Using intelligent transfection by NPs, we were able to induce apoptosis on EL4 cells and maintain acceptable SSC survival rates.
在癌症治疗中，通过纳米颗粒（NPs）进行的智能基因传递可靶向癌细胞，同时将对健康细胞的损伤降至最低。本研究评估了聚乳酸-乙醇酸共聚物（PLGA）-miR 143/206转染对小鼠白血病癌细胞（El4）和精原干细胞（SSCs）凋亡的影响。
在本实验研究中，使用了新生小鼠精原细胞和EL4癌细胞系。制备、表征了微小RNA-PLGA纳米颗粒并用叶酸进行靶向修饰。评估了几种剂量以获得合适的miR剂量，该剂量可诱导El4细胞发生适当凋亡，同时不损害精原干细胞。分别用每种miR的3种剂量处理细胞（对于miR 143，剂量为25、50和75 nmol；对于miR 206，剂量为50、100和150 nmol）。实验在24、48和72小时进行。通过MTT和膜联蛋白试剂盒研究细胞活力和凋亡情况。
根据MTT分析结果，miR 143的最佳剂量为75 nmol（精原干细胞为59.87±2.85%，EL4细胞为35.3±0.78%）（P≤0.05），对于miR 206，最佳剂量为150 nmol（精原干细胞为54.82±6.7%，EL4细胞为33.92±3.01%）（P≤0.05）。最佳时间为48小时。在这些剂量下，EL4细胞的存活率低于半数最大抑制浓度（IC），精原干细胞的存活率高于50%。膜联蛋白V染色也证实了所选剂量（对于miR 143，精原干细胞的总凋亡率为6.62%±1.8，EL4细胞为37.4%±4.2（P≤0.05），miR 206为（10.98%±1.5精原干细胞和36.4%±3.7 EL4细胞，P≤0.05）。
通过纳米颗粒进行智能转染，我们能够诱导EL4细胞凋亡并维持可接受的精原干细胞存活率。
