摘要
目的 研究长链非编码RNA含有WW和PDZ结构域2的膜相关鸟苷酸激酶反义RNA 3(LncRNA MAGI2-AS3)在鼻咽癌组织中的表达及与患者放射治疗(放疗)敏感性的关系,并探讨LncRNA MAGI2-AS3对鼻咽癌细胞放疗抵抗的影响及作用机制。方法 采用反转录荧光定量聚合酶链反应(RT-qPCR)检测LncRNA MAGI2-AS3在鼻咽癌组织中的表达水平,分析LncRNA MAGI2-AS3与鼻咽癌患者放疗敏感性的关系。常规培养鼻咽癌细胞CNE2,转染LncRNA MAGI2-AS3小干扰RNA(siRNA),根据是否转染siRNA分为si-NC组和si-LncRNA MAGI2-AS3组。采用CCK8实验检测各组细胞的放疗敏感性;放射处理后,采用平板克隆实验、流式细胞实验、裸鼠实验分别检测各组细胞克隆形成能力、细胞周期、凋亡情况和细胞体内生长能力;采用Western blot试验检测各组细胞周期蛋白D1(cyclin D1)、细胞周期蛋白依赖性激酶4(CDK4)、凋亡蛋白活化的含半胱氨酸的天冬氨酸蛋白水解酶(caspase)3、caspase9表达。结果 RT-qPCR检测结果显示,LncRNA MAGI2-AS3在鼻咽癌组织中表达上调(P<0.05)。LncRNA MAGI2-AS3高表达与患者放疗抵抗有关(P<0.05)。与si-NC组相比,si-LncRNA MAGI2-AS3组鼻咽癌细胞中LncRNA MAGI2-AS3表达减少、对放疗敏感性增加(P<0.05)。放射处理后,si-LncRNA MAGI2-AS3组鼻咽癌细胞克隆形成能力降低(P<0.05),细胞周期阻滞在G/G期(P<0.05),细胞凋亡增加(P<0.05),细胞体内生长能力显著降低(P<0.05),cyclin D1、CDK4表达降低,caspase3、caspase9表达增加(P<0.05)。结论 LncRNA MAGI2-AS3通过调控细胞周期和凋亡促进鼻咽癌细胞放疗抵抗,是逆转鼻咽癌放疗抵抗的潜在分子靶点。
Objective To study the expression of long chain noncoding RNA membrane associated guanylate kinase, WW and PDZ domain-containing 2 antisense RNA 3(LncRNA MAGI2-AS3) in nasopharyngeal carcinoma tissues and its relationship with the radiotherapeutic sensitivity of the patients, and to investigate the effect of LncRNA MAGI2-AS3 on nasopharyngeal carcinoma cell radiotherapeutic resistance and its action mechanism.Methods The reverse transcription fluorescent quantitative polymerase chain reaction(RT-qPCR) was used to detect the expression level of LncRNA MAGI2-AS3 in nasopharyngeal carcinoma tissues, and the relationship between the expression of LncRNA MAGI2-AS3 and the radiotherapeutic sensitivity in the patients with nasopharyngeal carcinoma.The nasopharyngeal carcinoma CNE2 cells were routinely cultured, transfected with LncRNA MAGI2-AS3 siRNA,and divided into the si-NC group and si-LncRNA MAGI2-AS3 group according to whether transfecting siRNA.The CCK8 experiment was used to detect the effect of cells in each group on the sensitivity of radiotherapy;after radiation treatment, the plate cloning experiment, flow cytometry experiment and nude mouse experiment were used to detect the cloning ability, cell cycle, apoptosis and growth ability of cells in vivo in each group;Western blot was used to detect the expression levels of cell cycle-related proteins D1( cyclin D1),CDK4,apoptosis-related proteins activated caspase3 and caspase9 in each group.Results The RT-qPCR detection results showed that the LncRNA MAGI2-AS3 expression was up-regulated in nasopharyngeal carcinoma tissues(P<0.05).The high expression of LncRNA MAGI2-AS3 was related with the radiotherapeutic resistance in the patients(P<0.05).Compared with the si-NC group, the expression level of LncRNA MAGI2-AS3 in nasopharyngeal carcinoma cells in the si-LncRNA MAGI2-AS3 group was decreased(P<0.05),and the radiotherapeutic sensitivity was increased(P<0.05).After radiation treatment, nasopharyngeal carcinoma cell clone formation ability in the si-LncRNA MAGI2-A
作者
戴润芝
徐莲
彭先兵
DAI Runzhi;XU Lian;PENG Xianbing(Department of ENT,Shiyan Municipal People's Hospital/Affiliated People's Hospital of Hubei Medical College,Shiyan,Hubei 442000,China)
出处
《国际检验医学杂志》
CAS
2022年第18期2232-2237,共6页
International Journal of Laboratory Medicine
基金
湖北省卫生健康委员会联合基金项目(WJ2019H437)。
关键词
鼻咽癌
长链非编码RNA含有WW和PDZ结构域2的膜相关鸟苷酸激酶反义RNA3
放疗抵抗
增殖
凋亡
nasopharyngeal carcinoma
long chain noncoding RNA membrane associated guanylate kinase
WW and PDZ domain-containing 2 antisense RNA 3
radiotherapeutic resistance
proliferation
apoptosis
