人脐带间充质干细胞分泌白细胞介素6促进骨肉瘤细胞增殖和迁移

hUC-MSCs promote proliferation and migration of osteosarcoma cells by secreting IL-6

目的:探讨人脐带间充质干细胞(hUC-MSCs)对骨肉瘤Saos-2细胞增殖和迁移的作用及分子机制。方法:组织块贴壁法分离培养hUC-MSCs,流式细胞术鉴定细胞表面标记物;CCK-8法和细胞计数法检测hUC-MSCs条件培养基(CM)、重组人白细胞介素6(rh IL-6)及IL-6中和抗体对Saos-2细胞增殖的作用;ELISA检测hUC-MSCs分泌IL-6的量;RT-PCR检测增殖相关基因增殖细胞核抗原(PCNA)、cyclin D1和survivin的转录水平;Transwell实验检测hUC-MSCs和Saos-2细胞迁移能力的变化。结果:hUC-MSCs可向Saos-2细胞迁移;hUC-MSCs-CM含有高浓度的IL-6,可达(1 835.5±134.1)ng/L;hUC-MSCs-CM和rhIL-6均能促进Saos-2细胞增殖和迁移,IL-6中和抗体能明显削弱hUC-MSCs-CM的促Saos-2细胞增殖和迁移作用;RT-PCR显示hUC-MSCs-CM和rhIL-6均能上调Saos-2细胞增殖相关基因PCNA、cyclin D1和survivin的表达,而IL-6中和抗体则削弱了这一作用。结论:脐带间充质干细胞能向骨肉瘤Saos-2细胞迁移,并通过分泌IL-6促进其增殖和迁移。

AIM： To investigate the effects of human umbilical cord-derived mesenchymal stem cells （hUC- MSCs） on the proliferation and migration of osteosarcoma cells （Saos-2） and the underlying molecular mechanism. METHODS： hUC-MSCs were isolated and cultured by tissue explants adherent method. The cell surface markers on hUC- MSCs were identified by flow cytometry. The effects of conditioned medium （CM） from hUC-MSCs （hUC-MSCs-CM）, re- combinant human interleukin-6 （rhIL-6） and IL-6 neutralizing antibody on the proliferation of Saos-2 cells were detected by CCK-8 assay and cell counting. IL-6 secretion of hUC-MSCs was assayed by ELISA. RT-PCR was used to assess the tran- scription level of proliferation-related genes proliferating cell nuclear antigen （PCNA）, cyclin D1 and survivin. The migra- tion potential of hUC-MSCs and Saos-2 cells was measured by Transwell assay. RESULTS ： hUC-MSCs migrated to Saos-2 cells, hUC-MSCs-CM contained a high concentration of IL-6, up to （1 835.5 -＋ 134. 1） ng/L. hUC-MSCs-CM and rhIL-6 promoted the proliferation and migration of Saos-2 cells. Addition of neutralizing antibody against IL-6 in the hUC-MSCs- CM impaired this proliferation and migration of Saos-2 ceils. The mRNA expression of PCNA, cyclin D1 and survivin was up-regulated by hUC-MSCs-CM and rhIL-6, while this effect was dramatically attenuated by treatment with IL-6 neutralizing antibody. CONCLUSION： hUC-MSCs migrate to osteosarcoma ceils and promote the proliferation and migration of osteo- sarcoma cells through secreting IL-6 in vitro.