阿帕替尼调控miR-129-5p/PBX3轴抑制食管癌细胞恶性生物学行为的研究  

Apatinib inhibits malignant biological behavior of esophageal cancer cells by modulation of miR-129-5p/PBX3 axis

作　　者：贺海发 万里新[2] 李凯 李寅[5] 王媛[3] 杜云辉 HE Haifa;WAN Lixin;LI Kai;LI Yin;WANG Yuan;DU Yunhui(Department of Pathology,Nanyang Central Hospital,Nanyang 473000,China;Department of Oncology,Nanyang Central Hospital,Nanyang 473000,China;Department of Gastrointestinal Oncology,Nanyang Central Hospital,Nanyang 473000,China;Zhang Zhongjing Institute of Chinese Medicine,Nanyang Institute of Technology,Nanyang 473004,China;Department of Pathology,Nanyang Medical College,Nanyang 473004,China)

机构地区：[1]南阳市中心医院病理科,南阳473000 [2]南阳市中心医院肿瘤内科,南阳473000 [3]南阳市中心医院消化道肿瘤内科,南阳473000 [4]南阳理工学院张仲景国医国药学院,南阳473004 [5]南阳医学高等专科学校病理学教研室,南阳473004

出　　处：《天津医科大学学报》2025年第2期126-133,共8页Journal of Tianjin Medical University

基　　金：河南省科技厅项目(242102310537)。

摘　　要：目的:探讨阿帕替尼(Apatinib)通过微小RNA(miR)-129-5p调控前B细胞白血病同源盒基因3(PBX3)对食管癌细胞恶性生物学行为的影响。方法:CCK-8法和划痕实验检测不同浓度Apatinib处理后KYSE450食管癌细胞的增殖、迁移能力;将KYSE450细胞分为KYSE450组、Apatinib-H组、Apatinib-H+miR-129-5p mimic组、Apatinib-H+miR-129-5p mimic+oe-PBX3组,qRT-PCR及免疫荧光检测miR-129-5p、PBX3表达水平,EdU、Transwell和TUNEL法检测细胞增殖、侵袭和凋亡情况,免疫印迹法检测Janus激酶2(JAK2)/信号转导及转录激活因子3(STAT3)信号通路相关蛋白水平。将293T人胚肾细胞分为293T+mimic NC组、293T+miR-129-5p mimic组,双荧光素酶报告基因实验验证miR-129-5p/PBX3的靶向关系。结果:20、40、60μmol/L的Apatinib均能抑制KYSE450细胞的增殖(t=4.416、11.331、13.121,均P<0.01)、迁移(t=2.277、4.286、6.722,均P<0.05);miR-129-5p与PBX3存在靶向关系;与KYSE450组相比,Apatinib-H组miR-129-5p mRNA水平升高(t=6.327,P<0.01),PBX3 mRNA及蛋白水平降低(t=6.098、10.403,均P<0.05);与Apatinib-H组相比,加入miR-129-5p模拟物可使KYSE450细胞的增殖率、侵袭个数及JAK2/STAT3通路关键蛋白水平降低,凋亡率升高(t=3.112、2.428、4.726、3.619、4.258,均P<0.05),过表达PBX3可逆转上述变化(t=3.698、3.199、4.082、3.563、5.840,均P<0.01)。结论:阿帕替尼可能通过上调miR-129-5p而靶向下调PBX3,抑制肿瘤细胞的恶性生物学行为。Objective:To investigate the effect of Apatinib on the malignant biological behavior of esophageal carcinoma cells by regulating the pre-B-cell leukemia homeobox gene 3(PBX3)through microRNA(miR)-129-5p.Methods:The CCK-8 assay and scratch assay were used to detect the proliferation and migration ability of KYSE450 esophageal cancer cells after treatment with different concentrations of Apatinib;KYSE450 cells were divided into the KYSE450 group,the Apatinib-H group,the Apatinib-H+miR-129-5p mimic group,the Apatinib-H+miR-129-5p mimic+oe-PBX3 group.The expression levels of miR-129-5p and PBX3 were detected by qRT-PCR and immunofluorescence.EdU,Transwell and TUNEL methods were used to detect cell proliferation,invasion and apoptosis,and the levels of proteins associated with the Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway were detected by Western blotting.The 293T cells were divided into 293T+mimic NC group and 293T+miR-129-5p mimic group,and Dual luciferase reporter gene assay was used to verifiy the miR-129-5p/PBX3 targeting relationship.Results:20,40,and 60μmol/L of Apatinib inhibited the proliferation(t=4.416,11.331,13.121,all P<0.01)and migration(t=2.277,4.286,6.722,all P<0.05)of the KYSE450 cells;there was a targeting relationship between miR-129-5p and PBX3.Compared with the KYSE450 group,miR-129-5p mRNA levels were increased(t=6.327,P<0.01),PBX3 mRNA and protein levels were decreased(t=6.098,10.403,both P<0.05)in the Apatinib-H group.Compared with Apatinib-H group,the addition of miR-129-5p mimics reduced the proliferation rate,the number of invasion and the level of key proteins of JAK2/STAT3 pathway,and increased the apoptosis rate of KYSE450 cells(t=3.112,2.428,4.726,3.619,4.258,all P<0.05),and overexpression of PBX3 reversed the above changes(t=3.698,3.199,4.082,3.563,5.840,all P<0.01).Conclusion:Apatinib may target down-regulate PBX3 by up-regulating miR-129-5p,and inhibit the malignant biological behavior of tumor cells.

关 键 词：食管癌 阿帕替尼 前B细胞白血病同源盒基因3 JANUS激酶2 信号转导及转录激活因子3 

分 类 号：R735.1[医药卫生—肿瘤]