目的:探讨Ig样结构域2黏附分子(adhesion molecule with Ig like domain 2,AMIGO2)在鼻咽癌(nasopharyngeal carcinoma,NPC)细胞增殖中的作用及其机制。方法:选用2017年9月至11月福建省肿瘤医院收集的10例NPC组织和10例正常鼻咽黏膜上...目的:探讨Ig样结构域2黏附分子(adhesion molecule with Ig like domain 2,AMIGO2)在鼻咽癌(nasopharyngeal carcinoma,NPC)细胞增殖中的作用及其机制。方法:选用2017年9月至11月福建省肿瘤医院收集的10例NPC组织和10例正常鼻咽黏膜上皮组织标本,以及NPC细胞系CNE-1、CNE-2、SUNE-1、6-10B、C666-1和人永生化鼻咽黏膜上皮细胞株NP69,用qPCR法检测NPC组织和细胞中AMIGO2 mRNA的表达。构建慢病毒载体干扰AMIGO2表达,用qPCR法验证其干扰效率;用CCK-8法、克隆形成及流式细胞术检测干扰AMIGO2表达对NPC细胞增殖、克隆形成和凋亡的影响,用Western blotting检测干扰AMIGO2表达对NPC细胞增殖及PI3K/AKT/mTOR信号通路相关标志蛋白表达的影响。结果:AMIGO2在NPC组织和CNE-2和SUNE-1细胞中高表达(均P<0.01)。慢病毒AMIGO2感染后,CNE-2和SUNE-1细胞的AMIGO2干扰效率均达50%以上。干扰AMIGO2表达,显著降低CNE-2和SUNE-1细胞增殖及克隆形成能力(均P<0.01)、明显提高细胞的凋亡率(均P<0.01);降低SUNE-1细胞中PI3K、AKT和mTOR磷酸化蛋白的表达水平(均P<0.01)、下调survivin和PCNA蛋白的表达水平(均P<0.01)。结论:AMIGO2通过激活PI3K/AKT/mTOR信号通路促进NPC细胞增殖并抑制其凋亡,提示AMIGO2可能是NPC治疗的潜在靶点。展开更多
The stiffness and nanotopographical characteristics of the extracellular matrix （ECM） influence numerous developmental, physiological, and pathological processes in vivo. These biophysical cues have therefore been a...The stiffness and nanotopographical characteristics of the extracellular matrix （ECM） influence numerous developmental, physiological, and pathological processes in vivo. These biophysical cues have therefore been applied to modulate almost all aspects of cell behavior, from cell adhesion and spreading to proliferation and differentiation. Delineation of the biophysical modulation of cell behavior is critical to the rational design of new biomaterials, implants, and medical devices. The effects of stiffness and topographical cues on cell behavior have previously been reviewed, respectively; however, the interwoven effects of stiffness and nanotopographical cues on cell behavior have not been well described, despite similarities in phenotypic manifestations. Herein, we first review the effects of substrate stiffness and nanotopography on cell behavior, and then focus on intracellular transmission of the biophysical signals from integrins to nucleus. Attempts are made to connect extracellular regulation of cell behavior with the biophysical cues. We then discuss the challenges in dissecting the biophysical regulation of cell behavior and in translating the mechanistic understanding of these cues to tissue engineering and regenerative medicine.展开更多
文摘目的:探讨Ig样结构域2黏附分子(adhesion molecule with Ig like domain 2,AMIGO2)在鼻咽癌(nasopharyngeal carcinoma,NPC)细胞增殖中的作用及其机制。方法:选用2017年9月至11月福建省肿瘤医院收集的10例NPC组织和10例正常鼻咽黏膜上皮组织标本,以及NPC细胞系CNE-1、CNE-2、SUNE-1、6-10B、C666-1和人永生化鼻咽黏膜上皮细胞株NP69,用qPCR法检测NPC组织和细胞中AMIGO2 mRNA的表达。构建慢病毒载体干扰AMIGO2表达,用qPCR法验证其干扰效率;用CCK-8法、克隆形成及流式细胞术检测干扰AMIGO2表达对NPC细胞增殖、克隆形成和凋亡的影响,用Western blotting检测干扰AMIGO2表达对NPC细胞增殖及PI3K/AKT/mTOR信号通路相关标志蛋白表达的影响。结果:AMIGO2在NPC组织和CNE-2和SUNE-1细胞中高表达(均P<0.01)。慢病毒AMIGO2感染后,CNE-2和SUNE-1细胞的AMIGO2干扰效率均达50%以上。干扰AMIGO2表达,显著降低CNE-2和SUNE-1细胞增殖及克隆形成能力(均P<0.01)、明显提高细胞的凋亡率(均P<0.01);降低SUNE-1细胞中PI3K、AKT和mTOR磷酸化蛋白的表达水平(均P<0.01)、下调survivin和PCNA蛋白的表达水平(均P<0.01)。结论:AMIGO2通过激活PI3K/AKT/mTOR信号通路促进NPC细胞增殖并抑制其凋亡,提示AMIGO2可能是NPC治疗的潜在靶点。
基金The authors would like to acknowledge funding support for Yong Yang from the National Science Foundation （CBET 1511759） and the National Institute of Health （NIH） （R15GM122953）, and for Kam W. Leong from NIH （HL109442, AI096305, GMl10494, and UH3 TR000505）, Guangdong Innovative and Entrepreneurial Research Team Program （2013S086）, and the Global Research Laboratory Program （Korean NSF GRL 2015032163）.
文摘The stiffness and nanotopographical characteristics of the extracellular matrix （ECM） influence numerous developmental, physiological, and pathological processes in vivo. These biophysical cues have therefore been applied to modulate almost all aspects of cell behavior, from cell adhesion and spreading to proliferation and differentiation. Delineation of the biophysical modulation of cell behavior is critical to the rational design of new biomaterials, implants, and medical devices. The effects of stiffness and topographical cues on cell behavior have previously been reviewed, respectively; however, the interwoven effects of stiffness and nanotopographical cues on cell behavior have not been well described, despite similarities in phenotypic manifestations. Herein, we first review the effects of substrate stiffness and nanotopography on cell behavior, and then focus on intracellular transmission of the biophysical signals from integrins to nucleus. Attempts are made to connect extracellular regulation of cell behavior with the biophysical cues. We then discuss the challenges in dissecting the biophysical regulation of cell behavior and in translating the mechanistic understanding of these cues to tissue engineering and regenerative medicine.
