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
目的构建LIMK1-siRNA真核表达载体,为骨关节发病机制的研究奠定基础。方法合成3条LIMK1的小RNA分子的寡聚脱氧核苷酸链,重组质粒测序,瞬时转染人软骨细胞,采用Western blot检测LIMK1表达情况。结果 Western blot检测LY3号质粒的细胞中LI...目的构建LIMK1-siRNA真核表达载体,为骨关节发病机制的研究奠定基础。方法合成3条LIMK1的小RNA分子的寡聚脱氧核苷酸链,重组质粒测序,瞬时转染人软骨细胞,采用Western blot检测LIMK1表达情况。结果 Western blot检测LY3号质粒的细胞中LIMK1蛋白表达明显下调,经通用引物对LY3号重组质粒测序鉴定,与设计序列核对完全相符。结论成功构建LIMK1-siRNA真核表达载体,有效沉默了人软骨细胞中LIMK1蛋白表达。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.
文摘目的构建LIMK1-siRNA真核表达载体,为骨关节发病机制的研究奠定基础。方法合成3条LIMK1的小RNA分子的寡聚脱氧核苷酸链,重组质粒测序,瞬时转染人软骨细胞,采用Western blot检测LIMK1表达情况。结果 Western blot检测LY3号质粒的细胞中LIMK1蛋白表达明显下调,经通用引物对LY3号重组质粒测序鉴定,与设计序列核对完全相符。结论成功构建LIMK1-siRNA真核表达载体,有效沉默了人软骨细胞中LIMK1蛋白表达。
