希佩尔-林道基因3996位腺苷到肌苷编辑与人表皮生长因子受体2阳性乳腺癌预后的相关分析  

Association between adenosine-to-inosine editing at 3996 position of von hippel lindau and prognosis of human epidermal growth factor receptor 2 positive breast cancer

作　　者：欧阳婷 陈仕桢 万舰[2] 刘丽 Ouyang Ting;Chen Shizhen;Wan Jian;Liu Li(Institute of Public Health of Guangzhou Medical University,Guangzhou 511436,China;Department of Galactophore,Guangdong Women and Children’s Hospital,Guangzhou 511400,China)

机构地区：[1]广州医科大学公共卫生学院,广州511436 [2]广东省妇幼保健院乳腺科,广州511400

出　　处：《中华实验外科杂志》2023年第4期626-629,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81871876)。

摘　　要：目的探讨希佩尔-林道(VHL)腺苷到肌苷编辑(ATIE)与人表皮生长因子受体2阳性(Her-2+)乳腺癌患者预后的关系及其机制。方法基于癌症基因组图谱数据库筛选与Her-2+乳腺癌患者预后关联的VHL ATIE。随机选取2014年至2019年广东省妇幼保健院相关患者81例,直接测序法检测ATIE位点的水平。采用荧光定量聚合酶链反应和蛋白印记法检测VHL表达。细胞计数试剂盒检测乳腺癌细胞耐药的半数抑制浓度(IC50)。生存分析采用Cox回归,组间比较采用t检验和方差分析,相关分析采用Spearman等级相关检验。结果数据库分析显示VHL转录本3996位腺苷到肌苷编辑(c.A3996I;HR=3.612,95%CI=1.093~11.940,P<0.05)、c.A3914I(HR=4.985,95%CI=1.442~17.230,P<0.05)与Her-2^(+)乳腺癌患者预后显著相关。直接测序法证实c.A3996I真实存在,低编辑患者组的死亡风险高于高编辑组(HR=3.980,95%CI=1.075~14.740,P<0.05),且淋巴结转移2/3期患者组c.A3996I的编辑水平显著低于淋巴结转移1期和无转移患者组[(9.923±5.741)%比(13.980±7.532)%和(17.820±9.424)%,F=5.140,P<0.01]。该位点编辑水平与增殖细胞核抗原表达(r=-0.282,P<0.05)呈负相关。c.A3996I编辑水平与VHL信使RNA(r=0.406,P<0.05)和蛋白(r=0.467,P<0.01)表达水平呈显著正相关,且在c.A3996I低编辑组,miR-143-5p与VHL的表达呈显著负相关(r=-0.145,P<0.05),但在高编辑组中两者无相关(r=0.051,P>0.05)。并且,耐曲妥珠单抗乳腺癌细胞VHL的表达显著低于正常细胞(0.540±0.116比1.011±0.195,t=4.159,P<0.01),未敲低VHL的乳腺癌细胞IC50低于敲低组(5.489μg/ml比22.120μg/ml,F=22.440,P<0.01),且敲低VHL的乳腺癌细胞迁移能力显著上升(1634±204比1140±252,t=3.049,P<0.05)。结论c.A3996I编辑可抑制VHL表达及其功能而影响Her-2+乳腺癌患者预后。Objective To explore the association of adenosine-inosine editing(ATIE)in von hippel lindau(VHL)with human epidermal growth factor receptor 2 positive(Her-2+)breast cancer prognosis and mechanism.Methods The cancer genome atlas database was used to sort the ATIE sites with effect on prognosis of Her-2+breast cancer patients.A total of 81 related patients were randomly recruited between 2014 to 2019 from the Guangdong Women and Children’s Hospital,and were used to determine the editing level of associated ATIE sites by direct sequencing.The fluorescent quantitative polymerase chain reaction and Western blotting were used to test the expression of VHL in breast cancer tissues.The cell counting kit-8 was applied for determining the half inhibitory concentration of cell resistance(IC50).Cox regression,t test and variance analysis,and Spearman rank correlation test were used for survival analysis,inter-group comparison,and correlation analysis,respectively.Results Database analysis showed that ATIEs at 3996 position of VHL transcript(c.A3996I;HR=3.612,95%CI=1.093-11.940,P<0.05),c.A3914I were significantly associated with prognosis of Her-2^(+)breast cancer patients(HR=4.985,95%CI=1.442-17.230,P<0.05).The c.A3996I was confirmed to be bona fide ATIE by direct sequencing,and the death rate in patients with hypo-editing of c.A3996I was higher than that in patients with hyper-editing(HR=3.980,95%CI=1.075-14.740,P<0.05).The cancer tissues from patients with stage 2-or 3-lymphatic metastasis exerted significantly lower editing of c.A3996I than those from patients with stage 1-lymphatic metastasis and without lymphatic metastasis[(9.923±5.741)%vs.(13.980±7.532)%and(17.820±9.424)%,F=5.140,P<0.01].The editing level was also correlated with Ki67 level(r=-0.282,P<0.05).Meanwhile,the editing level of c.A3996I was negatively correlated with mRNA and protein expression of VHL(r=0.406,P<0.05;r=0.467,P<0.01).In cancer tissues with low editing level,there was a negative correlation between microRNA-143-5p and VHL expression(r=-0.1

关 键 词：人表皮生长因子受体2 乳腺癌 腺苷 肌苷 

分 类 号：R737.9[医药卫生—肿瘤]