靶向抑制lncRNA CTB-191K22.5降低结直肠癌SW480细胞增殖和侵袭的机制研究

Targeted inhibition of lncRNA CTB-191K22.5 reduces the proliferation and invasion of colorectal cancer SW480 cells and its mechanism

目的探讨下调长链非编码RNA(lncRNA)CTB-191K22.5对结直肠癌SW480细胞增殖和侵袭的影响及分子机制。方法采用TCGA数据库分析结直肠癌组织和正常组织中CTB-191K22.5的表达差异。将CTB-191K22.5抑制物(Anti-CTB-191K22.5)和阴性抑制物(Control)转染至结直肠癌SW480细胞,分别记为观察组和对照组,实时定量聚合酶链式反应(qRT-PCR)评估抑制效果。MTT法和Transwell小室法分别评估SW480细胞的增殖和侵袭变化。Western blot评估SW480细胞中PI_(3)K/AKT/mTOR信号通路蛋白水平。生物信息学软件starBase v2.0预测CTB-191K22.5的靶基因。qRT-PCR评估SW480细胞中CTB-191K22.5靶基因的表达。正态分布的计量资料以均数±标准差(±s)表示,两组间比较采用t检验。结果与正常组织相比,结直肠癌组织中CTB-191K22.5呈现明显高表达(P<0.01)。对照组和观察组SW480细胞中CTB-191K22.5表达分别为6.60±0.85和1.08±0.21,转染Anti-CTB-191K22.5后CTB-191K22.5表达水平降低(P<0.01)。与对照组相比,观察组SW480细胞增殖能力下降(P<0.01)。对照组和观察组SW480细胞侵袭数分别为(135.4±16.29)个和(42.24±14.59)个,转染Anti-CTB-191K22.5后SW480细胞侵袭能力下降(P<0.01)。与对照组相比,观察组SW480细胞中PI_(3)K/AKT/mTOR信号通路蛋白表达水平降低。miR-326可能是CTB-191K22.5的靶基因。与对照组相比,转染Anti-CTB-191K22.5显著提高SW480细胞中miR-326表达水平(P<0.01)。结论结直肠癌组织中CTB-191K22.5高表达,下调CTB-191K22.5可能通过靶向miR-326抑制结直肠癌SW480细胞的增殖和侵袭。

Objective To explore the effect of down-regulation of long non-coding RNA(lncRNA)CTB-191K22.5 on the proliferation and invasion of colorectal cancer SW480 cells and the molecular mechanism.Methods The TCGA database was used to analyze the expression differences of CTB-191K22.5 in colorectal cancer tissues and normal tissues.The CTB-191K22.5 inhibitor(Anti-CTB-191K22.5)and negative inhibitor(Control)were transfected into colorectal cancer SW480 cells,denoted as Observation group and Control group,real-time quantitative polymerase chain reaction(qRT-PCR)was used to evaluate the inhibitory effect.MTT method and Transwell chamber method were used to evaluate the proliferation and invasion of SW480 cells.Western blot was used to evaluate the protein levels of PI_(3)K/AKT/mTOR signaling pathway in SW480 cells.The bioinformatics software starBase v2.0 was used to predict the target genes of CTB-191K22.5.qRT-PCR was used to evaluate the expression of CTB-191K22.5 target gene in SW480 cells.Measurement data were expressed as Mean±SD,and t-test was used for comparison between two groups.Results Compared with normal tissues,the expression of CTB-191K22.5 in colorectal cancer tissues was significantly increased(P<0.01).The expression of CTB-191K22.5 in SW480 cells of the Control group and Observation group were 6.60±0.85 and 1.08±0.21,respectively.The expression level of CTB-191K22.5 decreased after transfection with Anti-CTB-191K22.5(P<0.01).Compared with the Control group,the SW480 cell proliferation ability of the Observation group decreased(P<0.01).The invasion numbers of SW480 cells in the Control group and Observation group were(135.4±16.29)and(42.24±14.59),respectively.The invasion ability of SW480 cells decreased after transfection with Anti-CTB-191K22.5(P<0.01).Compared with the Control group,the expression levels of PI_(3)K/AKT/mTOR signaling pathway protein in SW480 cells in the Observation group decreased.miR-326 may be the target gene of CTB-191K22.5.Compared with the Control group,transfection with Anti-CTB-191K22.5 significantly increased the expression level of miR-326 in SW480 cells(P<0.01).Conclusion CTB-191K22.5 is highly expressed in colorectal cancer tissues,and down-regulation of CTB-191K22.5 may inhibit the proliferation and invasion of colorectal cancer SW480 cells by targeting miR-326.