摘要
目的:初步探究光动力疗法(Photodynamictherapy,PDT)对两种黑素瘤细胞迁移、侵袭的影响及作用机制。方法:取生长至对数期黑素瘤细胞A375和B16-F10,胰酶消化离心后计数后接种至六孔板内,培养使细胞融合度达到80%~90%以上后孵育光敏剂1 mmol/L 5-氨基酮戊酸(5-ALA)2 h后使用635 nm红光照射治疗,根据照射能量的不同分为0 J(对照组)、2.4 J(PDT 2.4J组)、4.8 J(PDT 4.8 J组)。采用划痕愈合实验和Transwell侵袭实验分别检测不同水平的PDT对B16-F10及A375系两种细胞迁移和侵袭能力的影响。采用蛋白质印迹法检测迁移和侵袭相关蛋白[Ki-67、血管内皮生长因子(Vascular endothelial growth factor,VEGF)、基质金属蛋白酶9(Matrix metalloproteinase 9,MMP9)]的相对表达量。结果:细胞划痕和Transwell实验的结果显示,PDT处理24 h后,相比对照组,PDT 2.4 J组及PDT 4.8 J组在B16-F10及A375两种细胞系中,划痕愈合率及侵袭细胞数目均明显减少(P<0.01);相比于PDT 2.4 J组,PDT 4.8 J组的划痕愈合率及侵袭细胞数目明显减少(P<0.01)。两PDT组在Ki-67、VEGF及MMP9三种蛋白的表达水平上均明显低于对照组(P<0.01);相比于PDT 2.4 J组,PDT 4.8 J组在三种蛋白的表达水平上更低(P<0.01)。结论:光动力治疗可显著抑制黑素瘤细胞系A375和B16-F10的侵袭与迁移能力,其机制可能是光动力治疗能够抑制促肿瘤生长相关因子的释放有关。
Objective A preliminary investigation of the effects and mechanisms of photodynamic therapy(PDT)on the migration and invasion of two types of melanoma cells.Methods Melanoma cells A375 and B16-F10,which grew to the log stage,were digested and centrifuged by pancreatic enzyme,counted,and inoculated into six-well plates.After culture,cell fusion reached more than 80%to 90%,the cells were incubated with photosensitizer 1 mmol/L 5-aminoketovalerate(5-ALA)for 2 h,and then treated with 635 nm red light irradiation.According to the different irradiation energy,it was divided into 0 J(control group),2.4J(PDT 2.4J group),4.8J(PDT 4.8J group).Scratch healing test and transwell invasion test were used to detect the effects of photodynamic therapy on the migration and invasion of melanoma cell lines A375 and B16-F10.Western blotting was used to detect the relative expression of migration and invasion-related proteins[Ki-67,vascular endothelial growth factor(VEGF),matrix metalloproteinase 9(MMP9)].Results The results of cell scratches and Transwell experiments showed that after 24 hours of PDT treatment,compared with the control group,the scratch healing rate and the invasion of cells in the PDT 2.4 J group and PDT 4.8 J group were in the B16-F10 and A375 cell lines.The number was significantly reduced(P<0.01),compared with the PDT 2.4 J group,the scratch healing rate and the number of invaded cells in the PDT 4.8 J group were significantly reduced(P<0.01).Compared with the PDT 2.4 J group,the PDT 4.8 J group had the expression levels of the three proteins is lower(P<0.01).Conclusion Photodynamic therapy can significantly inhibit the invasion and migration of melanoma cell lines A375 and B16-F10.The mechanism may be related to the inhibition of the expression of migration and invasion-related proteins.
作者
黎松
张敏
李蔼谕
吴亚光
LI Song;ZHANG Min;LI Aiyu;WU Yaguang(Department of Dermatology,the First Affiliated Hospital of Army Medical University(South West Hospital),Chongqing 400038,China;Department of Dermatology,People's Hospital of Shapingba District,Chongqing 400030,China)
出处
《中国美容医学》
CAS
2024年第2期56-59,共4页
Chinese Journal of Aesthetic Medicine
关键词
光动力疗法
黑素瘤细胞
迁移
侵袭
photodynamic therapy
melanoma cells
migration
invasion
