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 B7 family member B7-H3 is broadly expressed in many tissue and tumor types. B7-H3 expression is induced on some immune cells;however, its immunological function remains controversial, because both immunoenhancing ...The B7 family member B7-H3 is broadly expressed in many tissue and tumor types. B7-H3 expression is induced on some immune cells;however, its immunological function remains controversial, because both immunoenhancing and immunoinhibitory effects have been reported in human and mouse systems. We have previously reported the following: 1) murine B7-H3 specifically bound to Triggering receptor expressed on myeloid cells (TREM)-like transcript 2 (TLT-2, TREML2), a member of the TREM family of receptors;and 2) the B7-H3:TLT-2 pathway up-regulated T cell responses. However, the expression and function of human TLT-2 has not yet been clarified. A recent study found no evidence to support the existence of an interacttion between human B7-H3 and TLT-2. In this study, we demonstrated that human B7-H3 binds to TLT-2 and augments T cell responses. Human and mouse B7-H3Ig chimeric proteins cross-interacted with both human and mouse species of TLT-2-transduced cells. Human TLT-2 was expressed on freshly isolated, peripheral blood B cells and monocytes, and subpopulations of CD4+ and CD8+ T cells. Human TLT-2 expression on T cells did not correlate with na?ve or memory phenotypes and was diminished after culture, despite the presence of mitogenic stimuli. Constitutive TLT-2 expression on monocytes was also down-regulated after culture. Human B7-H3 transfectants augment IL-2 production from TLT-2-transduced T cell hybridomas, and IFN-γ production from peripheral blood CD4+ and CD8+ T cells. The enhanced responses were inhibited by the addition of anti-TLT-2 mAbs, suggesting TLT-2-mediated costimulatory effect. Our results demonstrate the existence of a functional interaction between human B7-H3 and TLT-2, and the restricted expression of TLT-2 on T cells and monocytes.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。
文摘The B7 family member B7-H3 is broadly expressed in many tissue and tumor types. B7-H3 expression is induced on some immune cells;however, its immunological function remains controversial, because both immunoenhancing and immunoinhibitory effects have been reported in human and mouse systems. We have previously reported the following: 1) murine B7-H3 specifically bound to Triggering receptor expressed on myeloid cells (TREM)-like transcript 2 (TLT-2, TREML2), a member of the TREM family of receptors;and 2) the B7-H3:TLT-2 pathway up-regulated T cell responses. However, the expression and function of human TLT-2 has not yet been clarified. A recent study found no evidence to support the existence of an interacttion between human B7-H3 and TLT-2. In this study, we demonstrated that human B7-H3 binds to TLT-2 and augments T cell responses. Human and mouse B7-H3Ig chimeric proteins cross-interacted with both human and mouse species of TLT-2-transduced cells. Human TLT-2 was expressed on freshly isolated, peripheral blood B cells and monocytes, and subpopulations of CD4+ and CD8+ T cells. Human TLT-2 expression on T cells did not correlate with na?ve or memory phenotypes and was diminished after culture, despite the presence of mitogenic stimuli. Constitutive TLT-2 expression on monocytes was also down-regulated after culture. Human B7-H3 transfectants augment IL-2 production from TLT-2-transduced T cell hybridomas, and IFN-γ production from peripheral blood CD4+ and CD8+ T cells. The enhanced responses were inhibited by the addition of anti-TLT-2 mAbs, suggesting TLT-2-mediated costimulatory effect. Our results demonstrate the existence of a functional interaction between human B7-H3 and TLT-2, and the restricted expression of TLT-2 on T cells and monocytes.
