AMIGO2通过激活PI3K/AKT/mTOR信号通路促进鼻咽癌细胞的增殖

AMIGO2 promotes proliferation of nasopharyngeal carcinoma cells by activating the PI3K/AKT/mTOR signaling pathway

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。

Objective:To explore the role of adhesion molecule with Ig like domain 2(AMIGO2)in the proliferation of nasopharyngeal carcinoma(NPC)cells and its mechanisms.Methods:A total of 10 NPC tissue samples and 10 normal nasopharyngeal epithelial tissue samples collected at Fujian Cancer Hospital during September 2017 and November 2017 were used for this study;in addition,NPC cell lines(CNE-1,CNE-2,SUNE-1,6-10B,C666-1)and human immobilized nasopharyngeal epithelial cell line NP69 were also collected.The relative expression ofAMIGO2 mRNAin above mentioned tissues and cell lines was detected by qPCR.Lentivirus vectors were constructed to interfere AMIGO2 mRNA expression,and qPCR was used to verify its interference efficiency.CCK-8 method,Clonal formation and Flow cytometry were performed to evaluate the effect of AMIGO2 interference on proliferation,clone formation and apoptosis of NPC cells.The influence of AMIGO2 interference on PI3K/AKT/mTOR signaling pathway and proliferation related molecular markers in NPC cells was assessed by Western blotting.Results:The results of qPCR showed that AMIGO2 was highly expressed in NPC tissues,CNE-2,and SUNE-1 cells(all P<0.01).The interference efficiency of AMIGO2 in CNE-2 and SUNE-1 cells could reach over 50%.The interfering of AMIGO2 expression significantly inhibited the proliferation and clone formation of CNE-2 and SUNE-1 cells(all P<0.01),promoted cell apoptosis(all P<0.01),reduced the phosphorylated protein expression levels of PI3K,AKT and mTOR in SUNE-1 cells(all P<0.01),as well as down-regulated the protein expressions of survivin and PCNA(all P<0.01).Conclusion:AMIGO2 may promote the proliferation as well as inhibit apoptosis of NPC cells by activating the PI3K/AKT/mTOR signaling pathway,suggesting that AMIGO2 may be a potential target for NPC therapy.