Growth-inhibitory effect of neurotrophin-3-secreting adipose tissue-derived mesenchymal stem cells on the D283-MED human medulloblastoma cell line

Abstract

Medulloblastoma (MBL), the most common malignant pediatric brain tumor, is incurable in about one-third of patients and can lead to long-term disabilities despite current multimodal treatments. The purpose of this study was to demonstrate in vitro biological effects of neurotrophins-3 (NT-3) on MBL cells and to evaluate the growth-inhibitory effect of neurotrophin-3 (NT-3)-secreting stem cells on tumor cells. We confirmed by western blotting that D283-MED cells express tyrosine kinase C, a specific receptor for NT-3. Analyzing the biological effects of NT-3 on MBL cells, we evaluated autophagy, apoptosis, senescence, and differentiation of tumor cells with NT-3. The NT-3 induced a concentration-dependent increase in apoptosis in the tumor cell line (P < 0.001). The high concentrations of NT-3 increased the expression of class III β-tubulin (P < 0.001) and decreased the expression of Nestin (P < 0.05). NT-3-secreting stem cells were produced by nucleofecting pIRES2.EGFP-NT3 into human adipose tissue-derived mesenchymal stem cells (hAT-MSCs) and their tropic property toward MBL cells was confirmed by migration assay. Double-layered co-culture experiments with the NT-3-secreting hAT-MSCs and D283-MED MBL cells were performed, and NT-3-induced cell death was studied by 3-(4,5-dimethylathiazol-2-yl)-2,5-dephenyl-tetrazolium bromide (MTT) assay. Consequently, the high concentrations of NT-3-secreting hAT-MSCs significantly (P < 0.05) increased the death of D283-MED cells in vitro. The present study demonstrated that both apoptotic cell death and neuronal differentiation of tumor cells were the mechanisms of growth-inhibitory effect of NT-3-secreting hAT-MSCs on MBL cell line.

Human-derived normal mesenchymal stem/stromal cells in anticancer therapies

“…AT-MSCs also suppress tumor growth and migrate to tumor tissues in tumor-bearing athymic mice 43. AT-MSCs have no growth-inhibitory effect on human medulloblastoma cell line D283-MED, however, MSCs transfected with neurotrophin-3 significantly inhibit the growth of D283-MED 44.…”

The leading edge: Emerging neuroprotective and neuroregenerative cell-based therapies for spinal cord injury

“…Another approach is the in vitro genetic modification of cells or the in vivo transfection of endogenous cells to secrete the necessary factors. For example, MSCs have been successfully engineered to express bFGF, 168 HGF, 169 NT3, 170 BDNF, 171 and GDNF 172 in vivo for various applications. SCs have also been transduced to overexpress BDNF and NT3 simultaneously 173 .…”

Adipose tissue stem cells: the great WAT hope

“…Similar bystander killing approaches have since been used, including use of thymidine kinase-expressing ASCs to facilitate ganciclovir-mediated tumor death [70]. Related studies have generated anti-tumorigenic ASCs that express oncolytic viruses [71], proapoptotic ligands [72–74] or anti-angiogenic effectors [73]. Importantly, these effects appear to target tumors specifically, with no adverse impact on either ASCs or normal host tissues.…”