环状RNA SLC41A2在非小细胞肺癌中的表达及其对细胞增殖、迁移和侵袭的影响

Expression of circSLC41A2 in non-small cell lung cancer and its influence on lung cancer cell proliferation,migration and invasion

目的探讨环状RNA SLC41A2(circSLC41A2/hsa_circ_0006260)在非小细胞肺癌(NSCLC)中的表达及临床意义, 探讨其对人NSCLC增殖、迁移和侵袭能力的影响。方法使用高通量环状RNA(circRNA)测序鉴定NSCLC组织中差异表达的circRNA, 并通过反转录实时荧光定量聚合酶链反应(RT-qPCR)进行验证, LSD-t检验分析其表达差异。收集临床数据, 采用χ^(2)检验分析circSLC41A2表达与各种临床病理特征的相关性。绘制受试者工作特征曲线(ROC)评价其诊断价值, 绘制Kaplan-Meier(K-M)生存曲线评价其预后评价价值。RT-qPCR检测人NSCLC细胞系(H1299、A549、HCC827、H1650、H359)和人正常肺上皮细胞系BASE-2B中circSLC41A2的表达, 采用单因素方差分析其表达差异。沉默circSLC41A2后, 采用细胞计数试剂盒(CCK-8)和集落形成实验检测增殖和集落形成, 流式细胞术检测细胞凋亡, 细胞划痕实验和Transwell实验评估迁移和侵袭能力, 采用LSD-t检验进行统计分析。结果 circSLC41A2在NSCLC中高表达[倍数差异(FC)为103.240, P<0.05]。癌组织circSLC41A2相对表达量高于癌旁组织(2.765±1.254比1.113±0.607, F=73.043, P<0.01)。癌组织中circSLC41A2的表达与肿瘤大小、淋巴结转移、TNM分期、表皮生长因子受体(EGFR)突变明显相关(χ^(2)=7.775、9.369、6.166、3.957, P<0.05)。circSLC41A2高表达组无进展生存期(PFS)显著低于低表达组。circSLC41A2具有良好的诊断价值[曲线下面积(AUC)=0.924, 95%可信区间(CI):0.870~0.987, P<0.01]。circSLC41A2在所有NSCLC细胞系中均高表达, 其中H1299的相对表达量最高。沉默circSLC41A2可显著抑制H1299细胞的增殖、侵袭和转移, 同时诱导细胞凋亡。结论 circSLC41A2在NSCLC组织和细胞系中高表达, 且与患者的临床病理特征相关。circSLC41A2在NSCLC中具有良好的诊断价值, 其低表达的患者比高表达的患者生存时间更长。沉默circSLC41A2可显著抑制H1299细胞的增殖、侵袭和迁移, 并促进细胞凋亡。

Objective To investigate the expression and clinical significance of circular RNA SLC41A2(circSLC41A2/hsa_circ_0006260)in non-small cell lung cancer(NSCLC)and assess its influence on the proliferation,migration,and invasion capabilities of human NSCLC.Methods High-throughput circular RNA(circRNA)sequencing identified differentially expressed circRNAs in NSCLC tissues,validated through quantitative reverse transcription polymerase chain reaction(RT-qPCR).LSD-t test was used to analyze the expression difference.Clinical data were gathered,and the correlation between circSLC41A2 expression and diverse clinicopathological characteristics was analyzed by Chi square test.The receiver operating characteristic(ROC)curve was plotted to assess diagnostic value,while the Kaplan-Meier(K-M)survival curve was constructed to evaluate its prognostic significance.RT-qPCR was used to assess the expression of circSLC41A2 in human NSCLC cell lines(H1299,A549,HCC827,H1650,H359)and the human normal lung epithelial cell line(BASE-2B).One way ANOVA was used to analyze the expression difference.Following circSLC41A2 silencing,proliferation and colony formation were assessed using cell counting kit-8(CCK-8)and colony formation assays,cell apoptosis was measured by flow cytometry,and migration and invasion abilities were evaluated using cell scratch and Transwell assays.LSD-t test was used for statistical analysis.Results CircSLC41A2 exhibited high expression in NSCLC[fold change(FC):103.240,P<0.05].In comparison to adjacent tissue,cancer tissue exhibited an elevated relative expression of circSLC41A2(2.765±1.254 vs.1.113±0.607,F=73.043,P<0.01).The CircSLC41A2 expression in cancer tissues was significantly correlated with tumor size,lymph node metastasis,TNM stage,and epidermal growth factor receptor(EGFR)mutation(χ^(2)=7.775,9.369,6.166,3.957,P<0.05).The progression free survival(PFS)in the high-expression group of circSLC41A2 was significantly lower than that in the low-expression group.CircSLC41A2 demonstrated good diagnostic value[area under curve(AUC)=0.924,95%confidence interval(CI):0.870-0.987,P<0.01].CircSLC41A2 exhibited high expression in all NSCLC cell lines,with H1299 displaying the highest relative expression.Silencing circSLC41A2 significantly inhibited the proliferation,invasion,and metastasis of H1299 cells,while inducing cell apoptosis.Conclusion CircSLC41A2 is up-expressed in NSCLC tissues and cell lines,which is correlated with the clinicopathological characteristics of patients.CircSLC41A2 demonstrates good diagnostic value in NSCLC,and patients with low expression experience longer survival than those with high expression.Silencing circSLC41A2 significantly inhibits the proliferation,invasion,and migration of H1299 cells,and promotes cell apoptosis.