Rab27A蛋白调控转录因子E盒结合锌指蛋白1对白细胞介素-6/信号转导与转录激活因子3信号通路的影响  

Mechanism of Rab27A regulating zinc finger E-box binding protein 1 mediated interleukin-6/signal transducer and activators of transcription 3 signaling pathway to promote the progression of triple-negative breast cancer

作　　者：杨树 范静婧 马斌林[1] Yang Shu;Fan Jingjing;Ma Binlin(Department of Breast and Thyroid Surgery,Xinjiang Medical University Afiliated Tumor Hospital,Urumqi 830011,China)

机构地区：[1]新疆医科大学附属肿瘤医院乳腺甲状腺外科,乌鲁木齐830011

出　　处：《中华实验外科杂志》2023年第4期611-614,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81960478)。

摘　　要：目的探索三阴性乳腺癌中Rab27A蛋白调控转录因子E盒结合锌指蛋白1(ZEB1)对白细胞介素(IL)-6/信号转导与转录激活因子3(STAT3)信号通路的影响。方法通过生信分析Rab27A与ZEB1在三阴性乳腺癌中表达,在体外培养三阴性乳腺癌细胞(MDA-MB-231、MDA-MB-468)使用慢病毒转染技术进行Rab27A、ZEB1过表达及敲减,并通过回复实验构建Rab27A过表达ZEB1敲减、Rab27A敲减ZEB1过表达的三阴性乳腺癌细胞系。运用蛋白质印迹法(Western blot)验证其蛋白表达。通过免疫印迹技术研究Rab27A调控ZEB1对IL-6/STAT3信号通路的影响,Shapiro-Wilk检测实验数据均服从正态分布。结果在三阴性乳腺癌MDA-MB-231细胞系中,蛋白质印迹法(Western blot)结果显示,ZEB1过表达组高于阴性对照组,差异有统计学意义(0.607±0.019比0.460±0.073,F=124.111,P<0.05),Rab27A敲减组低于阴性对照组,差异有统计学意义(0.118±0.003比0.460±0.073,F=124.111,P<0.05),ZEB1敲减组低于阴性对照组且有统计学意义(0.156±0.048比0.460±0.073,F=124.111,P<0.05)。在三阴性乳腺癌MDA-MB-468细胞系中,蛋白质印迹法(Western blot)结果显示Rab27A过表达组高于阴性对照组,差异有统计学意义(0.735±0.007比0.667±0.006,F=1859.845,P<0.05),ZEB1过表达组高于阴性对照组,差异有统计学意义(0.905±0.014比0.667±0.006,F=1859.845,P<0.05),Rab27A敲减组低于阴性对照组,差异有统计学意义(0.310±0.002比0.667±0.006,F=1859.845,P<0.05),ZEB1敲减组低于阴性对照组,差异有统计学意义(0.278±0.003比0.667±0.006,F=1859.845,P<0.05)。Rab27A过表达时ZEB1表达量高于ZEB1阴性对照组表达量,差异有统计学意义(0.735±0.007比0.905±0.014,F=1859.845,P<0.05),Rab27A敲减时ZEB1表达量低于ZEB1阴性对照组,差异有统计学意义(0.310±0.002比0.278±0.003,F=1859.845,P<0.05)。在三阴性乳腺癌MDA-MB-231细胞系中,Rab27A过表达时IL-6/STAT3信号通路被显著激活(2.010±0.092比1.440±0.089,F=196.692,P<0.05),三Objective To explore the effect of Rab27A protein regulating transcription factor zinc finger E-box binding protein 1(ZEB1)on interleukin(IL)-6/signal transducer and activators of transcription 3(STAT3)signaling pathway in triple-negative breast cancer.Methods The expression of Rab27A and ZEB1 in triple-negative breast cancer was analyzed by biogenic analysis.In the in vitro culture of triple-negative breast cancer cells(MDA-MB-231,MDA-MB-468),the overexpression and knockdown of Rab27A and ZEB1 were performed by lentivirus transfection technique.In addition,triple-negative breast cancer cell lines with Rab27A overexpression+ZEB1 knockdown and Rab27A knockdown+ZEB1 overexpression were constructed by response experiment.The protein expression was verified by Western blotting.The effect of Rab27A regulation on ZEB1 on IL-6/STAT3 signaling pathway was studied by Western blotting.Results In triple negative breast cancer MDA-MB-231 cell line,Western blotting results showed that ZEB1 overexpression group was higher than negative control group,with statistical significance(0.607±0.019 vs.0.460±0.073,F=124.111,P<0.05).Rab27A knockout group was lower than negative control group.The results were statistically significant(0.118±0.003 vs.0.460±0.073,F=124.111,P<0.05),and the ZEB1 knockout group was significantly lower than the negative control group(0.156±0.048 vs.0.460±0.073,F=124.111,P<0.05).In triple negative breast cancer cell line MDA-MB-468,Western blotting results showed that Rab27A overexpression group was higher than negative control group with statistical significance(0.735±0.007 vs.0.667±0.006,F=1859.845,P<0.05),and ZEB1 overexpression group was higher than negative control group.The Rab27A knockout group was significantly lower than the negative control group(0.310±0.002 vs.0.667±0.006,F=1859.845,P<0.05).ZEB1 knockout group was significantly lower than negative control group(0.278±0.003 vs.0.667±0.006,F=1859.845,P<0.05).The expression level of ZEB1 in Rab27A overexpression was higher than that in ZEB1

关 键 词：乳腺癌 白细胞介素-6 信号转导与转录激活因子3信号通路 

分 类 号：R737.9[医药卫生—肿瘤]