| Objective: IH is the most ordinary benign tumor in infancy.The endothelial cells that predominate in the proliferation period progressively decrease during the involution phase.Conversely,adipocytes increase and then ultimately dominate in the involution period.However,the mechanisms that are conducive to adipogenesis throughout spontaneous regression are unclear.Our research group have confirmed that IGF-1 can facilitate cell proliferation of Hem SCs and the differentiation of Hem SCs into adipocytes.Additionally,studies have confirmed that mi Rs may play a crucial role in regulating IGF-1R.We found that the expression of mi R-139-5p may be negatively associated with IGF-1R in Hem SCs via a pre-experiment.Our research was designed to investigate the mechanism of mi R-139-5p in Hem SCs.The current treatments for IH are limited,and mi R-139-5p might be a novel potential therapeutic target for IH.Methods:(1)Samples from twenty patients with infantile strawberry skin hemangioma in the proliferation phase who were under 1 year old were collected,cut and digested to obtain chylous suspension,and Hem SCs were selected by CD133 immunomagnetic beads for culture and passed to the third generation.(2)Hem SCs of the corresponding group were transfected with mi R-139-5p mimics NC,mi R-139-5p mimics,mi R-139-5p inhibitor NC and mi R-139-5p inhibitor,then cultured for 24 hours.(3)The expressions of mi R-139-5p and IGF-1R in the four groups were determined respectively by RT-q PCR and WB.(4)Plasmids containing IGF-1R-WT and IGF-1R-Mut were constructed respectively,and the relationship between mi R-139-5p and IGF-1R expression in the NC,IGF-1R-WT group and IGF-1R-Mut group were determined by luciferase reporter assay.(5)After transfection and culture for 24 hours,IGF-1(100ng/ml)was added to some groups for further treatment and culture for another 24 hours,getting mi R-139-5p mimics NC,mi R-139-5p mimics,mi R-139-5p NC,mi R-139-5p inhibitor,IGF-1,IGF-1+mi R-139-5p mimics,IGF-1 + mi R-139-5p inhibitor groups.(6)CCK-8 was used to determine the growth curves of Hem SCs at different times in seven groups.(7)Transwell migration assay was used to compare the migration ability of Hem SCs in the seven groups.(8)Oil red o staining was used to determine the number and presence of lipid droplets in the blank control group,IGF-1 group,IGF-1+mi R-139-5p mimics group and IGF-1+mi R-139-5p inhibitor group.(9)The expression of adipose conversion factors(C/EBP-?,C/EBP-?,PPAR-?)in Hem SCs of seven groups was determined respectively by RT-q PCR and WB.Results:(1)Overexpression of mi R-139-5p can reduce the expression of IGF-1R,while inhibition of mi R-139-5p can increase the expression of IGF-1R.(2)Dual luciferase reporter results verified that the IGF-1R is the mi R-139-5p's target gene.(3)Mi R-139-5p overexpression suppressed Hem SCs proliferation by targeting IGF-1R,while mi R-139-5p inhibition remarkably increased Hem SCs proliferation by targeting IGF-1R.(4)Mi R-139-5p overexpression remarkably suppressed Hem SCs migration by targeting IGF-1R,and mi R-139-5p inhibition remarkably increased Hem SCs migration.(5)Oil red o staining showed that IGF?1 treatment stimulated adipogenesis and lipid accumulation in Hem SCs.Additionally,mi R-139-5p overexpression suppressed IGF?1?induced lipid accumulation,whereas mi R-139-5p inhibition strengthened IGF?1?induced lipid accumulation.(6)RT-q PCR and WB analysis confirmed that IGF-1 could promote the expression of three adipose conversion factors in Hem SCs.mi R-139-5p overexpression inhibited the induction of IGF?1 on the expression of three adipose conversion factors,mi R-139-5p inhibition further enhanced the induction of IGF?1 on the expression of three adipose conversion factors.Conclusion: Mi R-139-5p can affect the IGF-1/IGF-1R pathway by regulating IGF-1R expression,which ultimately affects the proliferation,migration and adipogenesis of Hem SCs. |
PDF Full Text Request