摘要
目的探讨长链非编码RNA(lncRNA)DANCR调节微小RNA(miR)-33b/Zeste基因增强子同源物2(EZH2)轴对鼻咽癌细胞增殖和转移的影响。方法选取2023年5月至2024年10月在新乡医学院第一附属医院接受治疗的78例鼻咽癌患者为研究对象,经手术取鼻咽癌组织和癌旁组织样本,采用荧光定量聚合酶链反应(PCR)方法分析组织中lncRNA DANCR和miR-33b的表达水平;采用慢病毒转染建立lncRNA DANCR敲低和miR-33b过表达鼻咽癌细胞系,分别为lncRNA NC组、DANCR KD组、miRNA NC组、miR-33b OE组,采用细胞计数试剂盒(CCK-8)法检测鼻咽癌细胞的增殖能力;采用划痕实验和Transwell分析鼻咽癌细胞的迁移和侵袭能力。通过通过生物信息学软件预测、双荧光素酶报告基因和蛋白质免疫印迹技术分析lncRNA DANCR、miR-33b和EZH2的关系。组间比较采用t检验。结果癌旁组织lncRNA DANCR表达水平(1.07±0.17)显著低于鼻咽癌组织(1.65±0.20),差异有统计学意义(t=18.790,P<0.05)。癌旁组织miR-33b表达水平(0.95±0.22)显著高于鼻咽癌组织表达水平(0.60±0.15),差异有统计学意义(t=11.810,P<0.05)。lncRNA NC组细胞吸光度值、迁移数量和侵袭数量[1.25±0.10、(211.33±20.55)个、(86.67±8.6)个]明显高于DANCR KD组[0.83±0.18、(139.33±11.50)个、(49.67±8.02)个],差异有统计学意义(t=6.615、5.295、5.442,P<0.05)。miRNA NC组细胞吸光度值、迁移数量和侵袭数量[1.55±0.08、(203.67±17.01)个、(95.67±7.64)个]明显高于miR-33b OE组[1.04±0.09、(122.00±15.52)个、(54.00±6.56)个],差异有统计学意义(t=6.142、7.333、7.169,P<0.05)。lncRNA DANCR-WT/miR-33b模拟物转染组荧光素酶活性(0.44±0.05)显著低于lncRNA DANCR-Mut/miR-33b模拟物转染组(0.91±0.13),差异有统计学意义(t=5.728,P<0.05)。EZH2-WT/miR-33b模拟物转染组荧光素酶活性(0.67±0.06)显著低于EZH2-Mut/miR-33b模拟物转染组(1.01±0.10),差异有统计学意义(t=5.163,P<0.05)。lncRNA NC组细胞EZH2蛋白表达水平(1.11±0.16)显著高于DANCR KD组细胞(0.62±0.07),差异有统计学意义(t=4.748,P<0.05)。miRNA NC组细胞EZH2蛋白表达水平(1.00±0.12)显著高于miR-33b OE组细胞(0.52±0.08),差异有统计学意义(t=5.926,P<0.05)。结论lncRNA DANCR在鼻咽癌组织中呈高表达,通过调节miR-33b/EZH2轴,促进鼻咽癌细胞的增殖和转移等细胞生物学行为。
Objective To investigate the effect of long non-coding RNA(lncRNA)DANCR regulating the microRNA(miR)-33b/enhancer of zeste homolog 2(EZH2)axis on the proliferation and metastasis of nasopharyngeal carcinoma cells.Methods Totally,78 patients with nasopharyngeal carcinoma who received treatment in the First Affiliated Hospital of Xinxiang Medical University from May 2023 to October 2024 were selected as the research subjects.Nasopharyngeal carcinoma tissue and adjacent tissue samples were collected through surgery,and the expression levels of lncRNA DANCR and miR-33b in the tissues were analyzed using fluorescence quantitative polymerase chain reaction(PCR)method.The lentivirus transfection was done to establish lncRNA DANCR knockdown and miR-33b overexpression nasopharyngeal carcinoma cell lines,including lncRNA NC group,DANCR KD group,miRNA NC group,and miR-33b OE group,respectively.The proliferation ability of nasopharyngeal carcinoma cells was detected by cell counting kit-8(CCK-8)method.The scratch assay and Transwell analysis were used to investigate the migration and invasion ability of nasopharyngeal carcinoma cells.The relationship between lncRNA DANCR,miR-33b,and EZH2 was analyzed using bioinformatics software prediction,dual luciferase reporter genes,and protein immunoblotting techniques.The comparison of inter group measurement data was conducted using t-test.Results The expression level of lncRNA DANCR in adjacent cancer tissues(1.07±0.17)was significantly lower than that in nasopharyngeal carcinoma tissues(1.65±0.20)(t=18.790,P<0.05).The expression level of miR-33b in adjacent tissues(0.95±0.22)was significantly higher than that in nasopharyngeal carcinoma tissues(0.60±0.15)(t=11.810,P<0.05).The absorbance values,number of migrating cells,and number of invasive cells in lncRNA NC group[1.25±0.10,(211.33±20.55)cells,and(86.67±8.6)cells]were significantly higher than those in DANCR KD group[0.83±0.18,(139.33±11.50)cells,and(49.67±8.02)cells](t=6.615,5.295,5.442,P<0.05).The absorbance values,number of migrating cells,and number of invasive cells in the miRNA NC group were significantly higher than those in the miR-33b OE group[1.55±0.08,(203.67±17.01)cells,and(95.67±7.64)cells](t=6.142,7.333,7.169,P<0.05).The luciferase activity in the lncRNA DANCR-WT/miR-33b mimic transfection group(0.44±0.05)was significantly lower than that in the lncRNA DANCR-Mut/miR-33b mimic transfection group(0.91±0.13)(t=5.728,P<0.05).The luciferase activity in the EZH2-WT/miR-33b mimic transfection group(0.67±0.06)was significantly lower than that in the EZH2-Mut/miR-33b mimic transfection group(1.01±0.10)(t=5.163,P<0.05).The expression level of EZH2 protein in the lncRNA NC group(1.11±0.16)was significantly higher than that in the DANCR KD group(0.62±0.07)(t=4.748,P<0.05).The expression level of EZH2 protein in miRNA NC group(1.00±0.12)was significantly higher than that in miR-33b OE group(0.52±0.08)(t=5.926,P<0.05).Conclusion LncRNA DANCR is highly expressed in nasopharyngeal carcinoma tissues and promotes cell biology behaviors such as proliferation and metastasis by regulating the miR-33b/EZH2 axis.
作者
周航
王慧敏
余文发
崔粲
连荣
鲁保才
王银鑫
Zhou Hang;Wang Huimin;Yu Wenfa;Cui Can;Lian Rong;Lu Baocai;Wang Yinxin(Department of Otorhinolaryngology Head and Neck Surgery,the First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453100,China)
出处
《中华实验外科杂志》
2025年第7期1323-1326,共4页
Chinese Journal of Experimental Surgery
基金
2023年度河南省医学科技攻关计划项目(LHGJ20230516)。
