胚胎干细胞多能因子NANOG通过AMPK/mTOR通路介导乳腺癌细胞活性与侵袭  

Embryonic stem cell pluripotent factor NANOG mediates the activity and invasion of breast cancer cells via AMPK/mTOR pathway

作　　者：张开通[1] 张冰[1] 关山[1] Zhang Kaitong;Zhang Bing;Guan Shan(Breast Center,Beijing Tongren Hospital,Capital Medical University,Beijing 100176,China)

机构地区：[1]首都医科大学附属北京同仁医院乳腺中心,北京100176

出　　处：《中华内分泌外科杂志》2023年第2期156-161,共6页Chinese Journal of Endocrine Surgery

基　　金：首都医科大学附属北京同仁医院基金(2021-YJJ-PY-006)。

摘　　要：目的探讨胚胎干细胞多能因子NANOG通过AMPK/mTOR通路介导乳腺癌细胞活性与侵袭。方法2019年7月至2020年8月期间收集首都医科大学附属北京同仁医院58例乳腺癌患者,将每例患者入院时的临床资料进行对比分析。qRT-PCR检测癌旁组织与癌组织中NANOG的表达水平,Western blot验证NANOG调控AMPK/mTOR通路。使用敲减NANOG的特异性质粒或AMPK的抑制剂Compound C处理细胞,MTT检测细胞活力,Transwell检测细胞侵袭能力。结果qRT-PCR检测发现,与癌旁组织(1.00±0.31)相比,癌组织中NANOG表达水平(1.45±0.27)升高(t=8.34,P<0.001),与正常乳腺上皮细胞MCF-10A(1.00±0.12)相比,乳腺癌细胞系BT474(2.64±0.25,t=10.24,P=0.001)、MCF-7(1.56±0.13,t=5.48,P=0.005)、ZR-75-30(1.84±0.16,t=7.28,P=0.002)中NANOG表达水平均升高,在BT474细胞中表达最高,可作为预测乳腺癌的特异性生物分子。NANOG的表达水平可能与淋巴结转移、组织学分级、病理类型有关,与非淋巴结转移患者(1.36±0.23)或非浸润性患者(1.35±0.25)相比,淋巴结转移患者(1.54±0.27,t=2.61,P=0.012)或浸润性患者(1.53±0.26,t=2.60,P=0.012)中NANOG表达水平升高。敲减NANOG后AMPK蛋白及磷酸化水平升高,mTOR、p70S6K蛋白及磷酸化水平降低(均P<0.05)。细胞中敲减NANOG能抑制乳腺癌细胞活性(si-RNA:100.00±8.65,si-NANOG:58.36±4.58)(t=7.37,P=0.002)与侵袭能力(si-RNA:121.41±10.34,si-NANOG:58.34±8.41)(t=8.20,P=0.001),使用AMPK抑制剂处理细胞后,敲减NANOG产生的作用被解除(均P<0.05)。结论胚胎干细胞多能因子NANOG抑制AMPK/mTOR通路的激活可促进乳腺癌细胞活性与侵袭能力,NANOG可作为预测乳腺癌的有效生物分子。Objective To investigate the role of embryonic stem cell pluripotent factor NANOG in mediating the activity and invasion of breast cancer cells via AMPK/mTOR pathway.Methods A total of 58 breast cancer patients were collected from Jul.2019 to Aug.2020,and the clinical data of each patient at admission were collected for comparative analysis.qRT-PCR was used to detect the expression level of NANOG in adjacent tissues and cancer tissues,and Western blot was used to verify the regulation of AMPK/mTOR pathway by NANOG.Cells were treated with NANOG specific plasmid or AMPK inhibitor Compound C.Cell viability was detected by MTT and invasion ability was detected by Transwell.Results The expression of NANOG was increased in breast cancer tissues(adjacent to cancer tissue:1.00±0.31,cancer tissue:1.45±0.27,t=8.34,P<0.004)and cell lines(MCF-10A:1.00±0.12,BT474:2.64±0.25,t=10.24,P=0.001;MCF-7:1.56±0.13,t=5.48,P=0.005;ZR-75-30:1.84±0.16,t=7.28,P=0.002),which could be used as a specific biomolecule for predicting breast cancer(all P<0.05).The expression level of NANOG may be related to lymph node metastasis,histological grade and pathological type.Compared with patients with non-lymph node metastasis(1.36±0.23)or non-invasive patients(1.35±0.25),patients with lymph node metastasis(1.54±0.27,t=2.61,P=0.012)or invasive patients(1.53±0.26,t=2.60,P=0.012)had higher expression of NANOG.After NANOG knockdown,AMPK protein and phosphorylation levels were increased,while mTOR and p70S6K protein and phosphorylation levels were decreased(all P<0.05).Knockdown of NANOG in cells inhibited the activity and invasion of breast cancer cells(activity:si-RNA:100±8.65,si-NANOG:58.36±4.58,t=7.37,P=0.002;invasion:si-RNA:121.41±10.34,si-NANOG:58.34±8.41,t=8.20,P=0.001),and the effect of knockdown of NANOG was relieved after AMPK inhibitor was used in cells(all P<0.05).Conclusions Embryonic stem cell pluripotent factor NANOG promotes the activity and invasion of breast cancer cells by inhibiting the activation of AMPK/mTOR pathway.NANO

关 键 词：NANOG AMPK/mTOR 乳腺癌 细胞侵袭 

分 类 号：R737.9[医药卫生—肿瘤]