光动力治疗抑制黑色素瘤细胞迁移和侵袭能力的作用和机制

Inhibitory effects of photodynamics therapy on migratory and invasive abilities of melanoma cells and the mechanism

目的探查光动力疗法对黑色素瘤细胞系A375和B16-F10迁移、侵袭能力的影响,并探讨相关机制。方法采用划痕愈合实验和transwell侵袭实验分别检测光动力治疗对黑色素瘤细胞系A375和B16-F10迁移和侵袭能力的影响。实时定量PCR和ELISA检测黑色素瘤细胞自分泌炎症因子白介素-1(interleukin-1,IL-1)、白介素-6(interleukin-6,IL-6)、白介素-8(interleukin-8,IL-8)、转化生长因子-β(transforming growth factor-β,TGF-β)和肿瘤坏死因子(tumor necrosis factor-α,TNF-α)mRNA及上清分泌蛋白水平变化及其对PDT抑制黑色素瘤细胞迁移侵袭能力的影响。结果划痕愈合实验和transwell侵袭实验结果显示,在0 J、2.4 J和4.8 J剂量光动力治疗黑色素瘤细胞12 h与24 h后,黑色素瘤细胞迁移和侵袭能力明显降低(P<0.05);实时定量PCR和ELISA检测结果显示,光动力治疗对在2种细胞系中的IL-1、IL-8、IL-6和TNF-α水平影响不是非常一致;但是均能抑制TGF-β的表达(P<0.05),而且回补TGF-β(10 ng/ml)能显著逆转光动力治疗对2种细胞株迁移和侵袭能力的抑制作用。结论光动力治疗可显著抑制黑色素瘤细胞系A375和B16-F10的侵袭与迁移能力,其机制可能与其抑制了黑色素瘤自分泌TGF-β有关。

This study was designed to investigate the effects of photodynamics therapy(PDT)on the migratory and invasive abilities of melanoma cell lines A375 and B16-F10 and explore the underlying mechanism.Migratory and invasive abilities of A375 and B16-F10 were assessed respectively by the wound-healing and transwell assay after PDT.The mRNA expression and protein levels of interleukin-1(IL-1),interleukin-6(IL-6),interleukin-8(IL-8)and transforming growth factor-β(TGF-β),tumor necrosis factor-α(TNF-α)were analyzed using RT-qPCR and ELISA.Co-treatment of PDT and TGF-β(10 ng/ml)was performed to investigate whether supplement of TGFβcould reverse the inhibitory effects of PDT.The results of wound-healing and transwell assay showed that after treatment with 0 J,2.4 J and 4.8 J photodynamic therapy for 12 h and 24 h,the migratory and invasive abilities of melanoma cell lines were significantly reduced(P<0.05);the results of real-time quantitative PCR and ELISA showed that the expression level of TGF-βwas significantly decreased(P<0.05),while the expression levels of IL-1,IL-8,IL-6 and TNF-αwere not significantly changed in the two cell lines.Interestingly,after PDT combined 10 ng/ml TGF-βtreament,there was no significant difference in the abilities of migration and invasion.Taken together,PDT can inhibit the migration and invasion of melanoma A375 and B16-F10 by down-regulating autocrine TGF-β.