miR-10b靶向PARK2调控乳腺癌对PD1抑制剂的敏感性  

MiR-10b targeting PARK2 regulating the sensitivity of breast cancer cells to PD1 inhibitor

作　　者：刘金发 陈亚勤 温路生[1] 禹乐[1] 孟加榕[1] 陈艺锦 LIU Jin-fa;CHEN Ya-qin;WEN Lu-sheng;YU Le;MENG Jia-rong;CHEN Yi-jin(Department of Pathology,The 909th Hospital Logistic Support Forces(Southeast Affiliated Hospital of Xiamen University),Zhangzhou 363000,China;Department of Hematology,The 909th Hospital Logistic Support Forces(Southeast Affiliated Hospital of Xiamen University),Zhangzhou 363000,China)

机构地区：[1]联勤保障部队第九〇九医院(厦门大学附属东南医院)病理科,福建漳州363000 [2]联勤保障部队第九〇九医院(厦门大学附属东南医院)血液科,福建漳州363000

出　　处：《中国现代普通外科进展》2023年第1期13-17,23,共6页Chinese Journal of Current Advances in General Surgery

基　　金：联勤保障部队第909医院青年苗圃基金(18Y017,17Y009)。

摘　　要：目的:探讨mi R-10b靶向结合E3泛素蛋白连接酶(PARK2)对乳腺癌细胞应用程序性死亡蛋白1(PD1)抑制剂敏感性的影响。方法:回顾性收集联勤保障部队第九〇九医院2010年1月—2015年12月收治的146例乳腺癌患者的病例资料。PCR实验分析癌组织和癌旁组织中mi R-10b相对表达量;根据癌组织中mi R-10b表达水平中位数将146例乳腺癌患者分为mi R-10b高表达组和mi R-10b低表达组,绘制Kaplan-Meier生存曲线,分析癌组织中mi R-10b表达水平与患者预后相关性;荧光素酶实验分析mi R-10b与PARK2的关系,Person线性回归分析癌组织中mi R-10b表达水平与PARK2表达水平相关性;根据癌组织中PARK2表达水平中位数将146例乳腺癌患者分为PARK2高表达组和PARK2低表达组,Western blot方法分析PARK2低表达组与PD1蛋白水平性相关性;培养人乳腺癌细胞系,转染mi R-10b mimic(模拟物)使细胞中PARK2处于低表达,再通过转染mi R-10b mimic(模拟物)和mi R-10b inhibitor(抑制物)将细胞分为PARK2低表达+mi R-10b mimics组和PARK2低表达+mi R-10b inhibitor组,MTT法分析调控mi R-10b表达水平对乳腺癌细胞增殖能力和对PD1抑制剂敏感性的影响。结果:mi R-10b在乳腺癌组织中表达水平高于对应的癌旁组织(1.61±0.77比1.35±0.98,P<0.01);乳腺癌组织中mi R-10b高表达提示预后不良(P<0.01);mi R-10b靶向结合PARK2,乳腺癌组织中mi R-10b表达量与PARK2表达量呈负相关(r=-0.419,P<0.01);PARK2低表达组中PD1蛋白质表达较PARK2高表达组上调(灰度比值1.44±0.79比1.05±0.98,P<0.01);mi R-10b高表达使乳腺癌细胞系增殖能力上调(P<0.01),对PD1抑制剂敏感性下调(P<0.01)。结论:高表达mi R-10b提示乳腺癌预后不良,并降低对PD1抑制剂敏感性,机制可能与PARK2信号通路相关。Objective: To investigate the effect of Mir-10b targeting PARK2 binding on PD1inhibitor sensitivity in breast cancer cells. Methods: The clinical data of 146 breast cancer patients admitted to 909th hospital logistic support forces from January 2010 to December 2015 were retrospectively collected. The relative expression levels of Mir-10b and PARK2 and PD1 genes in cancer tissues and adjacent tissues were analyzed by q RT-PCR. According to the median expression level of mi R-10b in cancer tissues, 146 breast cancer patients were divided into mi R-10b high expression group and mi R-10b low expression group.Kaplan-Meier survival curves were drawn to analyze the correlation between the expression level of mi R-10b in cancer tissues and the prognosis of patients.Luciferase assay was used to analyze the relationship between mi R-10b and PARK2.Person linear regression analysis was used to analyze the correlation between mi R-10b expression level and PARK2 expression level in cancer tissues.According to the median PARK2 expression level in cancer tissues, 146 breast cancer patients were divided into PARK2 high expression group and PARK2 low expression group.The correlation between PARK2 low expression and PD1 protein level was analyzed by Western blot. Human breast cancer cell lines were cultured and transfected with mi R-10b mimic to reduce PARK2 expression.Transfected with mi R-10b mimic(mimic) and mi R-10b inhibitor(inhibitor), the cells were divided into PARK2 low expression +mi R-10b mimics group and PARK2low expression +mi R-10b inhibitor group. MTT assay was used to analyze the effect of regulating mi R-10b expression level on the proliferation ability and sensitivity to PD1 inhibitor of breast cancer cells. Results: Mir-10b expression level in breast cancer tissues was higher than that in adjacent tissues(1.61±0.77 vs 1.35±0.98, P<0.01). High expression of Mir-10b in breast cancer suggests poor prognosis(P<0.01). Mir-10b targets PARK2, and there is a negative correlation between Mir-10b expression and PARK2 ex

关 键 词：MIR-10B 乳腺肿瘤 E3泛素蛋白连接酶 程序性死亡受体1 预后 

分 类 号：R737.9[医药卫生—肿瘤]