长链非编码RNA SLCO4A1-AS1靶向微小RNA-615-5p对食管癌细胞增殖、凋亡和炎症因子表达的影响

Effect of long non-coding RNA SLCO4A1-AS1 targeting microRNA-615-5p on the proliferation,apoptosis and inflammatory factors expression in esophageal cancer cells

目的探讨长链非编码RNA(LncRNA)溶质载体有机阴离子转运蛋白家族成员4A1反义RNA1(SLCO4A1-AS1)靶向微小RNA-615-5p(miR-615-5p)对食管癌细胞增殖、凋亡和炎症因子表达的影响。方法运用实时荧光定量聚合酶链式反应(RT-qPCR)分析LncRNA SLCO4A1-AS1和miR-615-5p在食管癌组织、细胞系中表达情况。将si-NC、si-LncRNA SLCO4A1-AS1、miR-NC、miR-615-5p模拟物、pcDNA、pcDNA-LncRNA SLCO4A1-AS1、si-LncRNA SLCO4A1-AS1+anti-miR-NC、si-LncRNA SLCO4A1-AS1+anti-miR-615-5p分别转染Eca109细胞。CCK-8和流式细胞术用于检测细胞活力和凋亡率;平板克隆实验用于检测细胞增殖;酶联免疫吸附法(ELISA)试剂盒检测培养液中白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平。采用双荧光素酶报告基因法确定LncRNA SLCO4A1-AS1与miR-615-5p的关系。结果食管癌组织、细胞系中LncRNA SLCO4A1-AS1表达上调,miR-615-5p表达下调。抑制LncRNA SLCO4A1-AS1表达后,细胞活力、细胞克隆形成数量以及培养液中IL-6和TNF-α水平降低,miR-615-5p表达量、细胞凋亡率升高,差异有统计学意义(P<0.05)。与miR-NC组比较,miR-615-5p组Eca109细胞中miR-615-5p表达量、Cleaved-caspase-3蛋白水平、凋亡率升高,细胞活力、细胞克隆形成数量、培养液中IL-6和TNF-α水平降低,差异有统计学意义(P<0.05)。与si-LncRNA SLCO4A1-AS1+anti-miR-NC比较,si-LncRNA SLCO4A1-AS1+anti-miR-615-5p组Eca109细胞中miR-615-5p表达量、Cleaved-caspase-3蛋白水平、凋亡率降低,细胞活力、细胞克隆形成数量、培养液中IL-6和TNF-α水平升高,差异有统计学意义(P<0.05)。结论LncRNA SLCO4A1-AS1能够促进食管癌的发生和发展。抑制LncRNA SLCO4A1-AS1能够减少食管癌细胞的增殖,降低炎症因子的表达,诱导癌细胞凋亡。

Objective To investigate the effects of long non-coding RNA(LncRNA)solute carrier organic anion transporter family member 4A1(SLCO4A1-AS1)targeting microRNA-615-5p(miR-615-5p)in esophageal cancer cells on cell proliferation,apoptosis,and expression of inflammatory factors.Methods Real-time quantitative polymerase chain reaction(RT-qPCR)was used to analyze the expression of LncRNA SLCO4A1-AS1 and miR-615-5p in esophageal cancer tissues and cell lines.Eca109 cells were transfected with si-NC,si-LncRNA SLCO4A1-AS1,miR-NC,miR-615-5p mimic,pcDNA,pcDNA-LncRNA SLCO4A1-AS1,si-LncRNA SLCO4A1-AS1+anti-miR-NC,and si-LncRNA SLCO4A1-AS1+anti-miR-615-5p.Cell viability and apoptosis rate were measured by CCK-8 and flow cytometry,respectively;cell proliferation was determined by plate cloning assay;and levels of interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α)in the culture medium were measured using an enzyme-linked immunosorbent assay(ELISA)kit.A dual luciferase reporter gene assay was used to determine the relationship between LncRNA SLCO4A1-AS1 and miR-615-5p.Results Expression of LncRNA SLCO4A1-AS1 was upregulated,and miR-615-5p expression was downregulated in esophageal cancer tissues and cell lines.After inhibiting LncRNA SLCO4A1-AS1 expression,cell viability,the number of cell clones,and levels of IL-6 and TNF-αin the culture medium decreased,while miR-615-5p expression and apoptosis rate increased(P<0.05).Compared with the miR-NC group,the miR-615-5p group showed increased miR-615-5p expression,levels of Cleaved-caspase-3 protein,and apoptosis rate,and decreased cell viability,the number of cell clones,and levels of IL-6 and TNF-α in the culture medium(P<0.05).Compared with the si-LncRNA SLCO4A1-AS1+anti-miR-NC group,the si-LncRNA SLCO4A1-AS1+anti-miR-615-5p group showed decreased miR-615-5p expression,levels of Cleaved-caspase-3 protein,and apoptosis rate,and increased cell viability,the number of cell clones,and levels of IL-6 and TNF-α in the culture medium(P<0.05).Conclusion LncRNA SLCO4A1-AS1 can promote the occurrence and development of esophageal cancer.Inhibiting LncRNA SLCO4A1-AS1 can reduce the proliferation of esophageal cancer cells,decrease the expression of inflammatory factors,and induce apoptosis.