Qi Jin, Zhou Yali, Jiao Zuoyi, Wang Xu, Zhao Yang, Li Yangbin, Chen Huijuan, Yang Luxi, Zhu Hongwen, Li Yumin
The Second Hospital of Lanzhou University, Lanzhou, China.
Key Laboratory of Digestive System Tumors of Gansu Province, Lanzhou, China.
Cell Physiol Biochem. 2017;42(6):2242-2254. doi: 10.1159/000479998. Epub 2017 Aug 16.
BACKGROUND/AIMS: Mesenchymal stem/stromal cells (MSCs) are known to home to sites of tumor microenvironments where they participate in the formation of the tumor microenvironment and to interplay with tumor cells. However, the potential functional effects of MSCs on tumor cell growth are controversial. Here, we, from the view of bone marrow MSC-derived exosomes, study the molecular mechanism of MSCs on the growth of human osteosarcoma and human gastric cancer cells.
MSCs derived from human bone marrow (hBMSCs) were isolated and cultured in complete DMEM/F12 supplemented with 10% exosome-depleted fetal bovine serum and 1% penicillin-streptomycin, cell culture supernatants containing exosomes were harvested and exosome purification was performed by ultracentrifugation. Osteosarcoma (MG63) and gastric cancer (SGC7901) cells, respectively, were treated with hBMSC-derived exosomes in the presence or absence of a small molecule inhibitor of Hedgehog pathway. Cell viability was measured by transwell invasion assay, scratch migration assay and CCK-8 test. The expression of the signaling molecules Smoothened, Patched-1, Gli1 and the ligand Shh were tested by western blot and RT-PCR.
In this study, we found that hBMSC-derived exosomes promoted MG63 and SGC7901 cell growth through the activation of Hedgehog signaling pathway. Inhibition of Hedgehog signaling pathway significantly suppressed the process of hBMSC-derived exosomes on tumor growth.
Our findings demonstrated the new roles of hedgehog signaling pathway in the hBMSCs-derived exosomes induced tumor progression.
背景/目的：间充质干/基质细胞（MSCs）已知会归巢至肿瘤微环境部位，在那里它们参与肿瘤微环境的形成并与肿瘤细胞相互作用。然而，MSCs对肿瘤细胞生长的潜在功能影响存在争议。在此，我们从骨髓间充质干细胞衍生的外泌体角度，研究MSCs对人骨肉瘤和人胃癌细胞生长的分子机制。
分离人骨髓来源的间充质干细胞（hBMSCs），并在补充有10%去除外泌体的胎牛血清和1%青霉素-链霉素的完全DMEM/F12培养基中培养，收集含有外泌体的细胞培养上清液，并通过超速离心进行外泌体纯化。分别在有或没有Hedgehog信号通路小分子抑制剂的情况下，用hBMSC衍生的外泌体处理骨肉瘤（MG63）和胃癌（SGC7901）细胞。通过Transwell侵袭试验、划痕迁移试验和CCK-8试验测量细胞活力。通过蛋白质印迹法和RT-PCR检测信号分子Smoothened、Patched-1、Gli1和配体Shh的表达。
在本研究中，我们发现hBMSC衍生的外泌体通过激活Hedgehog信号通路促进MG63和SGC7901细胞生长。抑制Hedgehog信号通路显著抑制了hBMSC衍生的外泌体对肿瘤生长的作用。
我们的研究结果证明了Hedgehog信号通路在hBMSCs衍生的外泌体诱导肿瘤进展中的新作用。
