选定低氟浓度对人胶质瘤U-87MG细胞系迁移能力影响的测定。

Wojciech Żwierełło, Marta Skórka-Majewicz, Justyna Antoniewicz, Konrad Grzeszczak, Karolina Rogulska, Agnieszka Maruszewska
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引用次数: 0

摘要

氟(F)是一种属于卤素族的元素。少量的氟化物对于骨骼和牙齿的正常发育是必要的。然而,氟摄入量的增加和持续接触对人体机体有负面影响。最近的一些研究表明,氟化物影响了许多代谢途径,从理论上讲,这些代谢途径可能与许多类型癌症(包括脑肿瘤)的侵袭性发展有关。根据最近的研究,氟化物对癌细胞侵袭性的影响似乎很有可能,但实际上尚未探索。方法“伤口愈合”实验:在适当的NaF浓度(0.1-10µM)中传代72小时或3个月后,U-87MG细胞在6孔板(对照组+ NaF浓度)中生长。汇合(~ 80%)后,用200µl移液管尖划伤细胞层,并用PBS洗涤去除细胞碎片。在每孔中加入不含血清的新鲜培养基,在0,3,6,12和24小时用显微镜观察伤口愈合情况。细胞迁移试验在适当NaF浓度(0.1-10µM)下传代72小时或传代3个月后,将1 × 105个U-87MG细胞(对照+ 0.1-10µM NaF浓度,在含1%血清白蛋白牛种的无血清EMEM中)接种于孔径为8.0µM的24孔Transwell系统的上腔。将含有10%胎牛血清的EMEM加入下腔。孵育后,用棉签去除上表面的非迁移和非侵袭性细胞,用4%多聚甲醛固定下表面的细胞,并用吉氏染色法染色。在显微镜下拍照并计数细胞。结果在氟化钠的存在下,无论是短期培养还是长期培养,胶质母细胞瘤细胞的流动性都显著增加。重要的是,这种效果在最低浓度(0.1µM)下可见,并且在较高浓度(1-10µM)下增强。结论这些研究的结果可以为脑肿瘤患者的治疗方法提供新的思路,并引起人们对氟化物等环境因素的关注,这些因素可能已经阻碍了低剂量患者的治疗。考虑到在氟化物的影响下大脑中发生的许多过程,研究这种环境毒素对脑肿瘤进展和发展的影响似乎非常重要。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Determination of the effect of selected low fluoride concentrations on migratory abilities of human glioma U-87MG cell line.
IntroductionFluorine (F) is an element that belongs to the group of halogens. Small amounts of fluoride are necessary for the proper development of bones and teeth. However, increased intake of fluorine and continuous exposure has negative effects on the human organism. Some recent works have shown that fluoride affects many metabolic pathways that can theoretically be involved in the development of invasive potential in many types of cancers, including brain neoplasms. In light of recent studies, the influence of fluoride on the invasiveness of cancer cells seems highly probable but is practically unexplored. Methods„Wound healing” assayAfter 72 hours or three months of passaging in appropriate NaF concentrations (0.1-10 µM), U-87MG cells were grown in 6-well plates (controls + NaF concentrations indicated). After reaching confluence (~ 80%), the cell layers were scratched with 200 µl pipette tips and washed with PBS to remove cell debris. Fresh medium without serum was added to each well and the wound closure was visualized at 0, 3, 6, 12, and 24 hours using a microscope.Cell migration testAfter 72 hours or three months of passage in appropriate NaF concentrations (0.1-10 µM), a total of 1 × 105 U-87MG cells (controls + 0.1-10 µM concentration of NaF, in serum-free EMEM containing 1% serum albumin bovine species ) were inoculated in the upper chamber of a 24-well Transwell system with a pore size of 8.0 µm. EMEM containing 10% FBS was added to the lower chamber. After incubation, non-migrating and non-invasive cells on the upper surface were removed with a cotton swab and cells on the lower surface were fixed with 4% paraformaldehyde and stained with Giemsa. Photographs were taken and cells were counted under the microscope. ResultsOur observations showed that both in the case of short-term and long-term culture in the presence of sodium fluoride, the mobility of glioblastoma cells significantly increased. Importantly, the effect was visible at the lowest concentration (0.1 µM ) and increased at higher concentrations (1-10 µM) of NaF. ConclusionsThe results of these studies can shed new light on the therapeutic approach in people with brain tumors and draw attention to environmental factors such as fluoride, which may already hamper the treatment of patients at low doses. Considering the numerous processes taking place in the brain under the influence of fluoride, it seems extremely important to investigate the influence of this environmental toxin on the progression and development of brain tumors.
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