Although p21-activated kinase 2(PAK2)is an essential serine/threonine protein kinase,its role in the progression of lung squamous cell carcinoma(LUSC)has yet to be fully understood.We analyzed PAK2 mRNA levels,DNA cop...Although p21-activated kinase 2(PAK2)is an essential serine/threonine protein kinase,its role in the progression of lung squamous cell carcinoma(LUSC)has yet to be fully understood.We analyzed PAK2 mRNA levels,DNA copy numbers,and protein levels by quantitative reverse transcription-PCR and immunohistochemical staining in both human LUSC tissues and adjacent normal tissues.Then,we performed colony formation assays,cell counting kit-8 assays,Matrigel invasion assays,wound healing assays,and xenograft models in nude mice to investigate the functions of PAK2 in LUSC progression.We demonstrated that PAK2 mRNA levels,DNA copy numbers,and protein levels were upregulated in human LUSC tissues,compared with adjacent normal tissues.Additionally,higher PAK2 expression was associated with poorer prognosis in LUSC patients.In the in vitro study,we found that PAK2 promoted cell growth,migration,invasion,epithelialmesenchymal transition,and cell morphology regulation in LUSC cells.Mechanistically,PAK2 promoted tumor cell proliferation,migration,and invasion by regulating actin dynamics through the LIMK1/cofilin signaling pathway.Our findings indicate that the PAK2/LIMK1/cofilin signaling pathway may serve as a potential clinical marker and therapeutic target for LUSC.展开更多
基金National Natural Science Foundation of China(Grant No.32300615)Nanjing Medical Science and Technique Development Foundation(Grant No.JQX19010)。
文摘Although p21-activated kinase 2(PAK2)is an essential serine/threonine protein kinase,its role in the progression of lung squamous cell carcinoma(LUSC)has yet to be fully understood.We analyzed PAK2 mRNA levels,DNA copy numbers,and protein levels by quantitative reverse transcription-PCR and immunohistochemical staining in both human LUSC tissues and adjacent normal tissues.Then,we performed colony formation assays,cell counting kit-8 assays,Matrigel invasion assays,wound healing assays,and xenograft models in nude mice to investigate the functions of PAK2 in LUSC progression.We demonstrated that PAK2 mRNA levels,DNA copy numbers,and protein levels were upregulated in human LUSC tissues,compared with adjacent normal tissues.Additionally,higher PAK2 expression was associated with poorer prognosis in LUSC patients.In the in vitro study,we found that PAK2 promoted cell growth,migration,invasion,epithelialmesenchymal transition,and cell morphology regulation in LUSC cells.Mechanistically,PAK2 promoted tumor cell proliferation,migration,and invasion by regulating actin dynamics through the LIMK1/cofilin signaling pathway.Our findings indicate that the PAK2/LIMK1/cofilin signaling pathway may serve as a potential clinical marker and therapeutic target for LUSC.
文摘目的探索新生大鼠缺氧缺血脑损伤模型脑组织Cofilin1与ERK1/2蛋白表达与磷酸化及其在脑组织中分布的规律。方法新生7日龄SD大鼠,手术结扎左侧颈总动脉并经低氧处理造成缺氧缺血脑损伤(HIBD)模型,以假手术组为对照,应用western Blot和免疫组化方法检测损伤后一定时间段(0h～48h)损伤侧脑组织Cofilin1与磷酸化cofilin1(p-Cofilin1)及Erk1/2和磷酸化ERK1/2(p-ERK1/2)的表达变化,同时检测其在损伤大鼠脑组织和细胞中的表达和分布。结果 Western blot检测显示,HIBD新生SD大鼠脑组织中Cofilin1在HI后的表达与对照组相比也未见明显改变,但磷酸化Cofilin1在HI后1h～12h之间降低,24h后恢复正常;ERK1/2蛋白在不同时点的表达与对照组相比未见明显差异,但可见磷酸化ERK1/2(p-ERK1/2)在缺氧缺血处理(HI)后0h略低于正常,而在1-2h内迅速升高并高于正常对照和假手术,在2h后又逐渐降低。免疫组化结果显示Cofilin1与p-cofilin1在损伤大鼠脑中主要分布于海马和大脑皮质,尤其p-Cofilin表达分布以海马的颗粒细胞为主,并且在HI后7d-14d表达明显增强,在21d则明显降低。结论 Cofilin和ERK的磷酸化修饰与缺氧缺血脑损伤的发生和发展过程有较密切的关系,且cofilin1和p-cofilin1在大脑海马区的定位分布,提示其可能与大脑的学习记忆功能的损伤与修复有一定的关系。
