LINC00607通过抑制miR-372影响A549/DDP细胞的增殖和凋亡

LINC00607 affects the proliferation and apoptosis of A549/DDP cells through inhibiting miR-372

目的探讨LINC00607对肺癌顺铂耐药细胞系(A549/DDP)增殖、凋亡的影响和可能的分子机制。方法RT-qPCR检测肺癌组织、癌旁组织、A549、A549/DDP中LINC00607和miR-372的表达水平。将A549/DDP分为pcDNA3.1组、pcDNA3.1-LINC00607组、(pcDNA3.1-LINC00607+miR-NC)组、(pcDNA3.1-LINC00607+miR-372)组。采用四甲基偶氮唑蓝(MTT)法和集落形成实验检测细胞增殖;流式细胞测量术检测细胞凋亡;Western blot检测cleaved-caspase-3和Ki67蛋白表达;双荧光素酶报告基因和RT-qPCR实验确定LINC00607对miR-372的靶向调控作用。结果与癌旁组织比较,肺癌组织中LINC00607表达显著降低,miR-372表达显著升高(P<0.05)。与A549细胞比较,A549/DDP细胞中LINC00607表达显著降低,miR-372表达显著升高(P<0.05)。与pcDNA3.1组比较,pcDNA3.1-LINC00607组A549/DDP细胞存活率、克隆形成数、Ki67蛋白表达显著降低,而凋亡率、cleaved-caspase-3蛋白表达显著升高(P<0.05)。与(pcDNA3.1-LINC00607+miR-NC)组比较,(pcDNA3.1-LINC00607+miR-372)组A549/DDP细胞存活率、克隆形成数、Ki67蛋白表达显著升高,而凋亡率、cleaved-caspase-3蛋白表达显著降低(P<0.05)。LINC00607靶向负调控miR-372表达。结论LINC00607通过下调miR-372能够抑制A549/DDP细胞增殖,诱导细胞凋亡。

Objective To explore the effect of LINC00607 on proliferation and apoptosis of lung cancer cisplatin-resistant cell line(A549/DDP)and its possible molecular mechanism.Methods The expressions of LINC00607 and miR-372 in lung cancer tissues,adjacent tissues,A549,A549/DDP cells were detected by real-time fluorescence quantitative PCR(RT-qOCR).A549/DDP cells were divided into pcDNA3.1 group,pcDNA3.1-LINC00607 group,(pcDNA3.1-LINC00607+miR-NC)group,(pcDNA3.1-LINC00607+miR-372)group.Methyl thiazolyl tetrazolium(MTT)assacy and colony formation experiments were used to test cell proliferation;flow cytometry was used to measure apoptosis;and Western blot was used to detect the expression levels of cleared-caspase-3 and Ki67;Dual luciferase reporter gene and RT-qPCR experiments confirmed the targeted regulation effect of LINC00607 on miR-372.Results Compared with adjacent tissues,the expression of LINC00607 in lung cancer tissues was significantly reduced,while the expression of miR-372 was significantly increased(P<0.05).Comparedwith A549 cells,the expression of LINC00607 in A549/DDP cells was significantly reduced,while the expression of miR-372 was significantly increased(P<0.05).Compared with pcDNA3.1 group,A549/DDP cell survival rate,clone formation numbers,Ki67 protein expression in pcDNA3.1-LINC00607 group were significantly decreased,and apoptosis rate and cleared-caspase-3 protein expression were significantly increased(P<0.05).Compared with the(pcDNA3.1-LINC00607+miR-NC)group,the A549/DDP cell survival rate,clone formation,Ki67 protein expression in the(pcDNA3.1-LINC00607+miR-372)group were significantly increased,and the apoptosis rate,cleared-caspase-3 protein expression were significantly reduced(P<0.05).LINC00607 targeted at and negatively regulated miR-372 expression.ConclusionsLINC00607 inhibits proliferation and induces apoptosis of A549/DDP cells by down-regulating miR-372.