LINC00963调节微小RNA-506-3p/层粘连蛋白γ1轴对胃癌细胞增殖、迁移和侵袭的影响

Effects of LINC00963 regulating microRNA-506-3p/laminin gamma 1 axis on proliferation, migration and invasion of gastric cancer cells

目的探讨LINC00963调节微小RNA(miRNA,miR)-506-3p/层粘连蛋白γ1(LAMC1)轴对胃癌细胞增殖、迁移和侵袭的影响。方法选取2022年12月至2023年12月沧州市中心医院、华北医疗健康集团峰峰总医院和廊坊市人民医院收治的98例胃癌和癌旁组织样本作为研究对象,采用荧光定量聚合酶链反应(PCR)分析LINC00963和miR-506-3p的表达水平;采用慢病毒建立LINC00963敲低和miR-506-3p过表达细胞系,分别为长链非编码RNA(lncRNA)对照组、LINC00963 KD组、miRNA对照组、miR-506-3p组,分别采用细胞计数试剂测定细胞的增殖能力;采用划痕实验和Transwell分析细胞的迁移和侵袭能力。采用生物信息学和双荧光素酶报告基因分析LINC00963和miR-506-3p的关系。分析miR-506-3p靶基因,并采用蛋白质免疫印迹分析LINC00963和miR-506-3p对靶基因表达的影响。组间计量数据比较采用独立样本t检验。结果癌旁组织LINC00963表达水平(1.03±0.17)明显低于胃癌组织表达水平(1.65±0.15),差异有统计学意义(t=27.210,P<0.05)。癌旁组织miR-506-3p表达水平(0.92±0.20)明显高于胃癌组织表达水平(0.60±0.14),差异有统计学意义(t=12.760,P<0.05)。lncRNA对照组细胞吸光度值、迁移数量和侵袭数量[(1.71±0.09)、(87.67±5.47)个、(117.67±12.27)个]明显高于LINC00963 KD组细胞[(1.08±0.18)、(54.67±5.28)个、(86.50±6.95)个],差异有统计学意义(t=7.556、10.640、5.412,P<0.05)。miRNA对照组细胞吸光度值、迁移数量和侵袭数量[(1.87±0.07)、(81.67±9.63)个、(113.17±8.87)个]明显高于miR-506-3p组细胞[(1.13±0.09)、(60.67±8.04)个、(85.50±5.13)个],差异有统计学意义(t=16.070、5.596、6.605,P<0.05)。LINC00963和miR-506-3p存在结合位点。LAMC1是miR-506-3p的靶基因。lncRNA对照组细胞LAMC1蛋白表达水平(0.99±0.11)明显高于LINC00963 KD组细胞(0.53±0.07),差异有统计学意义(t=8.552,P<0.05)。miRNA对照组细胞LAMC1蛋白表达水平(1.08±0.08)明显高于miR-506-3p组细胞(0.48±0.09),差异有统计学意义(t=12.540,P<0.05)。结论LINC00963在胃癌组织中呈高表达,通过调节miR-506-3p/LAMC1轴,进而促进胃癌细胞的增殖、迁移和侵袭等细胞生物学行为。

ObjectiveTo investigate the effects of LINC00963 on the proliferation,migration and invasion of gastric cancer(GC)cells by regulating microRNA(miRNA,miR)-506-3p/laminin gamma 1(LAMC1)axis.MethodsTotally,98 GC and adjacent tissue samples from Cangzhou Central Hospital,Fengfeng General Hospital of North China Medical and Health Group and Langfang People’s Hospital from December 2022 to December 2023 were selected as the study objects,and the expression levels of LINC00963 and miR-506-3p were analyzed by fluorescence quantitative polymerase chain reaction(PCR).Lentivirus was used to establish LINC00963 knockdown(KD)cell lines and miR-506-3p overexpression cell lines,which were long non-coding RNA(lncRNA)control group,LINC00963 KD group,miRNA control group and miR-506-3p group,respectively.Cell counting reagent was used to determine cell proliferation ability.The migration and invasion ability of the cells was analyzed by scratch assay and Transwell assay.Bioinformatics and dual luciferase reporter gene were used to analyze the relationship between LINC00963 and miR-506-3p.Target genes of miR-506-3p were analyzed,and the effects of LINC00963 and miR-506-3p on the expression of target genes were analyzed by Western blotting.Independent samples t test was used to compare measurement data between groups.ResultsThe expression level of LINC00963 in adjacent tissues(1.03±0.17)was significantly lower than that in GC tissues(1.65±0.15,t=27.210,P<0.05).The expression level of miR-506-3p in adjacent tissues(0.92±0.20)was significantly higher than that in GC tissues(0.60±0.14,t=12.760,P<0.05).The absorbance value,migration number and invasion number of lncRNA control group[(1.71±0.09),(87.67±5.47)cells and(117.67±12.27)cells]were significantly increased as compared with those of LINC00963 KD group[(1.08±0.18),(54.67±5.28)cells,(86.50±6.95)cells,t=7.556,10.640,5.412,P<0.05].The absorbance value,migration number and invasion number of cells in miRNA control group[(1.87±0.07),(81.67±9.63)cells,(113.17±8.87)cells]were significantly greater than those in miR-506-3p group[(1.13±0.09),(60.67±8.04)cells,(85.50±5.13)cells,t=16.070,5.596,6.605,P<0.05].The LAMC1 protein expression level in lncRNA control group(0.99±0.11)was significantly higher than that in LINC00963 KD group(0.53±0.07,t=8.552,P<0.05).The LAMC1 protein expression level in miRNA control group(1.08±0.08)was significantly higher than that in miR-506-3p group(0.48±0.09,t=12.540,P<0.05).ConclusionLINC00963 is highly expressed in GC tissues,and can promote the proliferation,migration and invasion of GC cells by regulating the miR-506-3p/LAMC1 axis.