放射性皮肤损伤中细胞干性的转变OA

OBJECTIVE:To investigate changes in cellular stemness transition in radiation-induced skin injury in cell culture.METHODS:Human immortalized keratinocytes(HaCaT)were exposed to X-ray at 0,5,10,and 20 Gy.Cell migration ability at 24 h and 48 h post-irradiation was assessed using the scratch wound healing assay and transwell migration assay.Western blot was used to detect the expression of stemness-related proteins KLF4,c-myc and LGR5 in HaCaT cells at 24 h and 48 h after irradiation.In the animal experiments,the skin of the right hind limb of SD rats was irradiated with a single dose of 20 Gy X-ray.At 15 and 35 days post-irradiation,western blot was performed to detect the expression of stemness-related proteins SOX2,Nanog,KLF4,c-myc,and LGR5 in the irradiated skin cells.A non-irradiated control group(0 Gy)was included,with 4 rats per group.RESULTS:Compared with the control group(0 Gy),the in vitro scratch wound healing assay at 48 h post-irradiation showed that cell migration ability was significantly reduced in the 5,10,and 20 Gy groups(P＜0.05).In contrast,the transwell assay demonstrated significantly enhanced migration ability in all irradiated groups(P＜0.01),with the 10 Gy group still retaining a certain level of migration capacity.At 24 h post-irradiation,KLF4,c-myc and LGR5 protein expression levels in HaCaT cells were significantly decreased in all irradiated dose groups(P＜0.01).At 48 h post-irradiation,KLF4,c-myc and LGR5 protein expression levels were significantly increased(P＜0.01).In the animal experiments,compared with the control group(0 Gy),at 15 days after 20 Gy X-ray irradiation,expression levels of stemness-related proteins SOX2,c-Myc,and LGR5 in rat skin were significantly elevated(all P＜0.01).At 35 days post-irradiation,expression levels of SOX2,Nanog,and LGR5 were significantly decreased(P＜0.05 or P＜0.01).CONCLUSION:In radiation-induced skin injury,cellular sternness transition capacity expressed dynamic changes which might cause inhibition of repair,promotion of regeneration with gradual decline in the long term.