LINC01929对非小细胞肺癌细胞增殖、迁移、侵袭和β-catenin通路的影响

Influence of LINC01929 on proliferation,migration,invasion of non-small cell lung cancer cell and β-catenin signaling pathway

目的探讨长基因间非蛋白编码RNA 1929(LINC01929)对非小细胞肺癌(NSCLC)增殖、迁移和侵袭的调控作用,及其对β-catenin信号通路活性的影响。方法从Starbase数据库获取LINC01929在肺腺癌和肺鳞癌组织中的表达情况,并分析其与患者预后的关系。收集2017年6月至2020年12月手术切除的NSCLC组织和癌旁组织标本各50例。采用实时荧光定量PCR(qPCR)法检测LINC01929在NSCLC组织和肺上皮细胞BEAS-2B以及NSCLC细胞株H1975、H1299、SPC-A-1、H460和A549中的表达。将A549细胞分为空白对照组(未转染)、阴性对照组(转染siRNA-NC)和LINC01929干扰组(转染siRNA-LINC01929)。CCK-8法、划痕愈合实验和Transwell小室实验检测细胞增殖、迁移和侵袭情况。qPCR和Western blotting法检测各组细胞β-catenin信号通路相关因子β-连环蛋白(β-catenin),波形蛋白(Vimentin)和基质金属蛋白酶-7(MMP-7)的表达。结果生物信息学分析显示,LINC01929在肺腺癌和肺鳞癌组织中的表达水平均明显上调(P<0.01);LINC01929高表达肺腺癌患者的总生存时间明显低于LINC01929低表达患者(P<0.05)。qPCR检测结果显示,LINC01929在NSCLC组织中的相对表达量(1.436±0.082)高于癌旁组织(1.000±0.055),差异有统计学意义(P<0.01)。LINC01929在NSCLC细胞株中的相对表达量亦高于正常肺上皮细胞株BEAS-2B(P<0.01),选择表达水平最高的A549细胞株(2.120±0.127)进行后续实验。转染siRNA-LINC01929后,LINC01929干扰组LINC01929的表达水平显著降低(P<0.01)。与阴性对照组比较,LINC01929干扰组细胞增殖活力、划痕愈合率和穿膜细胞数量均显著降低(P<0.01)。LINC01929干扰组中β-catenin、Vimentin和MMP-7的mRNA和蛋白表达水平均低于阴性对照组(P<0.01)。结论LINC01929在NSCLC组织和细胞株中表达上调,干扰LINC01929的表达抑制了NSCLC细胞的增殖、迁移和侵袭,其调控机制可能与β-catenin信号通路失活有关。

Objective To investegate the function of long intergenic non-protein coding RNA 1929(LINC01929)on proliferation,migration and invasion of non-small cell lung cancer(NSCLC),and its effect onβ-catenin signaling pathway.Methods The expression of LINC01929 in lung adenocarcinoma and lung squamous cell carcinoma tissues was obtained from the Starbase database,and its relationship with prognosis of patients was analyzed.Fifty pairs of paracancerous tissues and NSCLC tissues surgically removed from June 2017 to December 2020 were collected.The expression of LINC01929 in NSCLC tissues and cell lines(H1975,H1299,SPC-A-1,H460 and A549)were determined by real time-quantitative PCR(qPCR).A549 cells were divided into blank control group(no transfection),negative control group(transfection of siRNA-NC)and siRNA-LINC01929 group(downregulation of LINC01929).CCK-8,scratch and transwell assays were conducted to detect the cell proliferation,migration and invasion.Western blotting and qPCR were applied to detect the expressions ofβ-catenin,Vimentin and matrix metallopeptidase-7(MMP-7).Results Bioinformatics analysis showed that the expression level of LINC01929 was significantly upregulated in lung adenocarcinoma and lung squamous cell carcinoma tissues(P<0.01).The overall survival of lung adenocarcinoma patients with high expression of LINC01929 was significantly lower than that of patients with low expression of LINC01929(P<0.05).The results of qPCR showed that the relative expression of LINC01929 in NSCLC tissues(1.436±0.082)was higher than that in adjacent tissues(1.000±0.055),and the difference was statistically significant(P<0.01).The relative expression of LINC01929 in NSCLC cell line was also higher than that of normal lung epithelial cell line BEAS-2B(P<0.01),and the A549 cell line with the highest expression level(2.120±0.127)was selected for subsequent experiments.After transfection with siRNA-LINC01929,the expression level of LINC01929 in the LINC01929 interference group was significantly reduced(P<0.01).Compared with the negative control group,the cell proliferation viability,scratch healing rate and number of penetrating cells in the LINC01929 interference group were significantly reduced(P<0.01).The expression levels ofβ-catenin,Vimentin and MMP-7 mRNA and protein in the LINC01929 interference group were lower than those in the negative control group(P<0.01).Conclusion The expression of LINC01929 is upregulated in NSCLC tissues and cell lines,and interference with the expression of LINC01929 inhibits the proliferation,migration and invasion of NSCLC cells,and its regulatory mechanism may be related to the inactivation of theβ-catenin signaling pathway.