circ-ERBB2通过抑制miR-136-5p维持乳腺癌中的HER2下游信号促进曲妥珠单抗耐药  

circ-ERBB2 promotes trastuzumab resistance by inhibiting miR-136-5p to maintain HER2 downstream signal in breast cancer

作　　者：王兰 董江萌 雷勇 齐义新[3] WANG Lan;DONG Jiangmeng;LEI Yong;QI Yixin(The First of Department of General Surgery,Hengshui People's Hospital,Hebei Hengshui 053099,China;Department of Glandular Vascular Surgery,Hengshui People's Hospital,Hebei Hengshui 053099,China;Department of Oncology,the Fourth Hospital of Hebei Medical University,Hebei Shijiazhuang 053011,China)

机构地区：[1]衡水市人民医院普外一科,河北衡水053099 [2]衡水市人民医院腺体血管外科,河北衡水053099 [3]河北医科大学第四医院肿瘤科,河北石家庄053011

出　　处：《现代肿瘤医学》2023年第7期1195-1201,共7页Journal of Modern Oncology

基　　金：河北省重点研发计划项目(编号:19277799D);河北省衡水市科技计划项目(编号:2021014088Z)。

摘　　要：目的:探讨circ-ERBB2/miR-136-5p轴是否参与曲妥珠单抗耐药乳腺癌的耐药性分子机制。方法:qPCR法检测曲妥珠单抗敏感和耐药乳腺癌组织,以及HER2^(-)和HER2^(+)乳腺癌组织中circ-ERBB2和miR-136-5p的表达水平。建立小鼠异种移植瘤模型,评估敲低/过表达circ-ERBB2联合过表达/敲低miR-136-5p对乳腺癌细胞成瘤性及对曲妥珠单抗耐药性的影响。生物信息学和双荧光素酶报告基因分析circ-ERBB2与miR-136-5p的序列互作位点。CCK-8细胞增殖能力测定敲低/过表达circ-ERBB2联合过表达/敲低miR-136-5p以及给予曲妥珠单抗治疗对于乳腺癌常规或曲妥珠单抗耐药BT-474(Trast-resist)细胞增殖能力的影响。Western blot法检测敲低/过表达circ-ERBB2联合过表达/敲低miR-136-5p后,对常规或BT-474(Trast-resist)细胞内HER2、p-Akt、p-ERK1/2表达水平的影响。结果:与曲妥珠单抗敏感乳腺癌组织相比,曲妥珠单抗耐药乳腺癌组织中circ-ERBB2的表达水平显著升高,miR-136-5p表达水平显著降低(P<0.01)。无论激素受体表达为阳性或是阴性,与HER2^(-)乳腺癌组织相比,在HER2^(+)乳腺癌组织中circ-ERBB2表达显著上调,miR-136-5p表达水平被显著抑制(P<0.01)。敲低circ-ERBB2的表达或过表达miR-136-5p均能够抑制BT-474(Trast-resist)细胞增殖及其胞内HER2、p-Akt和p-ERK1/2的表达水平(P<0.01)。结论:circ-ERBB2能够通过抑制miR-136-5p参与维持乳腺癌中的HER2下游信号并促进曲妥珠单抗耐药。Objective:To explore whether the circ-ERBB2/miR-136-5p axis is involved in the molecular mechanism of drug resistance in trastuzumab-resistant breast cancer.Methods:The expression levels of circ-ERBB2 and miR-136-5p in trastuzumab-sensitive and resistant breast cancer tissues,as well as in HER2-and HER2+breast cancer tissues were detected by qPCR.Xenograft mouse model was established to evaluate the effects of knockdown/overexpression of circ-ERBB2 combined with overexpression/knockdown of miR-136-5p on tumorigenicity and trastuzumab resistance of breast cancer cells.Bioinformatics and dual-luciferase reporter analysis were used to analyze the interaction site between circ-ERBB2 and miR-136-5p.CCK-8 cell proliferation assay was used to analyze the proliferation ability of BT-474 or BT-474(Trast-resist)cells,when knockdown/overexpressed circ-ERBB2 was combined with overexpressed/knockdown miR-136-5p and cells were administered with Trastuzumab.Western blot was used to detect the expression levels of HER2,p-Akt and p-ERK1/2 in BT-474 or BT-474(Trast-resist)cells after knockdown/overexpression of circ-ERBB2 was combined with overexpression/knockdown of miR-136-5p.Results:Compared with trastuzumab-sensitive breast cancer tissues,the expression level of circ-ERBB2 in trastuzumab-resistant breast cancer tissues was significantly increased,and the expression level of miR-136-5p was significantly decreased(P<0.01).Regardless of whether hormone receptor expression was positive or negative,compared with HER2^(-) breast cancer tissues,the expression of circ-ERBB2 was significantly upregulated and the expression level of miR-136-5p was significantly inhibited( P <0.01) in HER2^(+) breast cancer tissues.Knockdown of circ-ERBB2 expression or overexpression of miR-136-5p could inhibit the proliferation of BT-474(Trast-resist) cells and the expression levels of HER2,p-Akt and p-ERK1/2 in cells( P <0.01). Conclusion: circ-ERBB2 can maintain HER2 downstream signaling in breast cancer and promote trastuzumab resistance by inhibitin

关 键 词：HER2阳性乳腺癌 曲妥珠单抗耐药 circ-ERBB2 miR-136-5p 

分 类 号：R73-3[医药卫生—肿瘤]