过表达DNALI1抑制低氧环境中鼻咽癌细胞株CNE-2z侵袭与迁移

DNALI1 inhibits invasion and migration of nasopharyngeal carcinoma CNE-2z cell in a hypoxic environment

目的探讨模拟肿瘤内低氧微环境下,动力蛋白轴突轻链中间体1(DNALI1)对鼻咽癌细胞株CNE-2z增殖、迁移、侵袭和凋亡的影响。方法R语言软件分析GEO数据库中鼻咽癌组织的3个数据集,分析其差异基因(DEGs)。低氧是肿瘤微环境基本特征,体外细胞研究均在低氧工作站(1﹪O_(2))中进行,以模拟体内低氧微环境。将DNALI1过表达质粒和空载体质粒包装成慢病毒后感染CNE-2z细胞,构建DNALI1基因稳定过表达的CNE-2z细胞株。将CNE-2z细胞分别进行正常培养(空白对照,BC),感染空载体慢病毒(阴性对照,NC)和感染过表达DNALI1慢病毒(过表达DNALI1,DNALI1-OE)。实时荧光定量聚合酶链式反应(RT-qPCR)和Western blot检测DNALI1 mRNA和蛋白表达水平;集落形成实验检测细胞增殖能力;Transwell实验检测细胞迁移和侵袭能力;流式细胞术检测细胞凋亡水平。多组间比较采用单因素方差分析,组间两两比较采用LSD-t检验。结果生物信息学分析发现,与健康人鼻咽组织相比,鼻咽癌组织中DNALI1基因低表达。在低氧环境中,BC组与NC组细胞增殖、凋亡、迁移和侵袭比较,差异均无统计学意义;与BC组和NC组比较,DNALI1-OE组细胞增殖能力和凋亡水平比较,差异均无统计学意义(P均>0.05)。与BC组和NC组比较,DNALI1-OE组细胞迁移能力[(198.67±3.22),(199.00±3.00)比(75.00±7.55)个]和侵袭能力[(240.67±16.01),(259.67±3.06)比(83.33±9.50)个]降低,差异具有统计学意义(P均<0.001)。结论低氧环境中,DNALI1过表达可抑制CNE-2z细胞侵袭与迁移能力,但对细胞增殖、凋亡无明显影响。

Objective To investigate the effects of dynein axonemal light intermediate chain 1(DNALI1)on proliferation,invasion,migration and apoptosis of CNE-2z cells in a hypoxic environment.Methods Three data sets about nasopharyngeal carcinoma tissues in Gene Expression Omnibus(GEO)database were analyzed to find differential expressedgenes(DEGs)through R language software.Hypoxia is the characteristic of tumor microenvironment.This study was carried out in a hypoxia work station(1﹪O_(2))to simulate the hypoxic microenvironment in vivo.Stable overexpressed DNALI1CNE-2z cells were established by infecting with lentivirus packaged with DNALI1 overexpressed plasmid.CNE-2z cells were cultured normally(blank control,BC),infected with empty vector virus(negative control,NC)and DNALI1 overexpression vector virus(DNALI1-OE)respectively,and then exposed to hypoxic environment(1﹪O2).The mRNA and protein expression levels of DNALI1 gene were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blot.The proliferation of CNE-2z cells was measured by colony formation assay.Transwell assay was used to detect the migration and invasion of CNE-2z cells.The apoptotic cells of CNE-2z cells were detected by flow cytometry.One-way ANOVA was used for comparison among multiple groups.LSD-t test was used for pairwise comparison between groups.P values of less than 0.05 were considered statistically significant.Results Through bioinformatics analysis,it was found that the expression of DNALI1 was downregulated in nasopharyngeal carcinoma compared with normal nasopharyngeal tissue.After DNALI1 overexpression,the proliferation ability and apoptotic cells of CNE-2z cells remained unchanged in the hypoxic environment compared with BC and NC groups(all P>0.05).However,compared with BC group and NC group,the cell migration ability(198.67±3.22),(199.00±3.00)vs(75.00±7.55)and invasion ability(240.67±16.01),(259.67±3.06)vs(83.33±9.50)were significantly inhibited in DNALI1-OE group(all P<0.001).Conclusion DNALI1 overexpression inhibits the invasion and migration of CNE-2z cells in hypoxic environment,but has no effect on proliferation and apoptosis of CNE-2z cells.