[目的]探究REC8(Meiotic recombination protein 8,REC8)调控PAK1对三阴性乳腺癌(Triple-negative breast cancer,TNBC)转移活性的影响。[方法]收集36例TNBC患者的癌组织及癌旁组织,免疫组化分析REC8在TNBC组织及癌旁组织中阳性表达率。...[目的]探究REC8(Meiotic recombination protein 8,REC8)调控PAK1对三阴性乳腺癌(Triple-negative breast cancer,TNBC)转移活性的影响。[方法]收集36例TNBC患者的癌组织及癌旁组织,免疫组化分析REC8在TNBC组织及癌旁组织中阳性表达率。将TNBC MDA-MB-231细胞分为3组(2.5×10^(5)个/m L):阴性对照组(转染100nmol/孔、50 nmol/L阴性对照siRNA)、pc DNA REC8组(转染2μg/孔、1μg/μL pc DNA REC8过表达质粒)、siPAK1组(转染100 nmol/孔、50 nmol/L siPAK1)。通过Ed U染色实验分析MDA-MB-231细胞增殖活性,Transwell实验分析MDA-MB-231细胞的侵袭、迁移能力,蛋白免疫印迹实验分析MDA-MB-231细胞中REC8、PAK1蛋白的表达。[结果]与癌旁组织比较,REC8在TNBC组织中表达降低(P<0.05)。与阴性对照组相比,pc DNA REC8、siPAK1组的MDA-MB-231细胞增殖能力下降(P<0.05);pc DNA REC8、siPAK1组的MDA-MB-231细胞的侵袭、迁移能力下降(P<0.05);pc DNA REC8、siPAK1组的MDA-MB-231细胞PAK1蛋白表达降低(P<0.05)。[结论]REC8在TNBC组织中表达降低。上调TNBC MDA-MB-231细胞的REC8表达能够通过抑制PAK1进而减弱MDA-MB-231细胞的增殖、侵袭、迁移活性。展开更多
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
文摘[目的]探究REC8(Meiotic recombination protein 8,REC8)调控PAK1对三阴性乳腺癌(Triple-negative breast cancer,TNBC)转移活性的影响。[方法]收集36例TNBC患者的癌组织及癌旁组织,免疫组化分析REC8在TNBC组织及癌旁组织中阳性表达率。将TNBC MDA-MB-231细胞分为3组(2.5×10^(5)个/m L):阴性对照组(转染100nmol/孔、50 nmol/L阴性对照siRNA)、pc DNA REC8组(转染2μg/孔、1μg/μL pc DNA REC8过表达质粒)、siPAK1组(转染100 nmol/孔、50 nmol/L siPAK1)。通过Ed U染色实验分析MDA-MB-231细胞增殖活性,Transwell实验分析MDA-MB-231细胞的侵袭、迁移能力,蛋白免疫印迹实验分析MDA-MB-231细胞中REC8、PAK1蛋白的表达。[结果]与癌旁组织比较,REC8在TNBC组织中表达降低(P<0.05)。与阴性对照组相比,pc DNA REC8、siPAK1组的MDA-MB-231细胞增殖能力下降(P<0.05);pc DNA REC8、siPAK1组的MDA-MB-231细胞的侵袭、迁移能力下降(P<0.05);pc DNA REC8、siPAK1组的MDA-MB-231细胞PAK1蛋白表达降低(P<0.05)。[结论]REC8在TNBC组织中表达降低。上调TNBC MDA-MB-231细胞的REC8表达能够通过抑制PAK1进而减弱MDA-MB-231细胞的增殖、侵袭、迁移活性。
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.
