Department of Pediatric Surgery, Nihon University School of Medicine, 30-1 Oyaguchi-Kamicho, Tokyo, 173-8610, Japan.
Department of Functional Morphology, Division of Cell Regeneration and Transplantation, Nihon University School of Medicine, Tokyo, 173-8610, Japan.
Pediatr Surg Int. 2022 Dec 22;39(1):58. doi: 10.1007/s00383-022-05304-x.
Mesenchymal stem cells (MSCs) can induce differentiation of neuroblastoma (NB) cells. Properties of dedifferentiated fat cells (DFATs) are similar to those of MSCs. Here, we investigated whether DFATs can induce NB cell differentiation and suppress cell proliferation.
DFATs were obtained from mature adipocytes isolated from adipose tissue from a ceiling culture. NB cells were cultured in a medium with or without DFATs and, subsequently, cultured in a DFAT-conditioned medium (CM) with or without phosphatidylinositol 3-kinase (PI3K) inhibitor. The neurite lengths were measured, and mRNA expression levels of the neurofilament (NF) and tubulin beta III (TUBβ3) were assessed using quantitative real-time RT-PCR. Cell viability was assessed using the WST-1 assay.
NB cells cultured with DFATs caused elongation of the neurites and upregulated the expression of NF and Tubβ3. NB cells cultured in DFAT-CM demonstrated increased cell viability. NB cells cultured with DFAT-CM and PI3K inhibitors suppressed cell viability. NB cells cultured with DFAT-CM and PI3K inhibitor demonstrated increased neurite length and expression, and upregulation of Tubβ3.
The combined use of DFAT-CM and PI3K inhibitors suppresses the proliferation of NB cells and induces their differentiation. Thus, DFAT may offer new insights into therapeutic approaches in neuroblastoma.
间充质干细胞(MSCs)可诱导神经母细胞瘤(NB)细胞分化。去分化脂肪细胞(DFATs)的特性与 MSCs 相似。本研究旨在探讨 DFATs 是否能诱导 NB 细胞分化并抑制细胞增殖。
DFATs 从脂肪组织中分离的成熟脂肪细胞经天花板培养获得。NB 细胞在含有或不含有 DFATs 的培养基中培养,然后在含有或不含有磷脂酰肌醇 3-激酶(PI3K)抑制剂的 DFAT 条件培养基(CM)中培养。测量神经突的长度,并通过定量实时 RT-PCR 评估神经丝(NF)和微管β III(TUBβ3)的 mRNA 表达水平。使用 WST-1 测定法评估细胞活力。
与 DFAT 共培养的 NB 细胞导致神经突伸长,并上调 NF 和 Tubβ3 的表达。在 DFAT-CM 中培养的 NB 细胞表现出更高的细胞活力。与 PI3K 抑制剂共培养的 NB 细胞在 DFAT-CM 中抑制细胞活力。在 DFAT-CM 和 PI3K 抑制剂中培养的 NB 细胞表现出更长的神经突长度和表达增加,以及 Tubβ3 的上调。
DFAT-CM 和 PI3K 抑制剂的联合使用可抑制 NB 细胞的增殖并诱导其分化。因此,DFAT 可能为神经母细胞瘤的治疗方法提供新的思路。