长链非编码RNA ABHD11-AS1促进胃癌细胞糖酵解并加速肿瘤恶性进展

Long non-coding RNA ABHD11-AS1 promotes glycolysis in gastric cancer cells to accelerate tumor progression

目的探讨长链非编码RNA ABHD11-AS1对胃癌细胞糖酵解的影响及其作用的分子机制。方法分别将空载质粒pcDNA-Vector和过表达质粒pcDNA-ABHD11-AS1转染入ABHD11-AS1低表达的MKN45和MGC803胃癌细胞中,构建过表达组(pcDNA-ABHD11-AS1)和空载质粒对照组胃癌细胞株。运用CCK-8法检测细胞的增殖活性并绘制生长曲线,克隆形成实验检测细胞克隆形成能力,Transwell检测细胞迁移、侵袭能力,葡萄糖摄取实验及乳酸生成实验检测细胞糖酵解水平的变化;LncMAP数据库分析查找ABHD11-AS1可能调控的转录因子,查阅文献进行分析挑选候选转录因子,Western blot明确ABHD11-AS1是否影响候选转录因子的表达量。结果与对照组相比,转染pcDNA-ABHD11-AS1后,MGC803和MKN45胃癌细胞中ABHD11-AS1基因表达量明显上升(P<0.01),CCK8和成克隆实验表明细胞增殖加快(P<0.05),克隆形成能力增强,Tanswell实验结果证实细胞迁移(11±2 vs 27±3;17±4 vs 28±3,P<0.01)、侵袭(15±3 vs 26±2;10±1 vs 35±2,P<0.01)作用增强;上调ABHD11-AS1后,胃癌细胞葡萄糖摄取及乳酸生成增多(P<0.05)。分析数据库结果显示ABHD11-AS1可能调控经典糖酵解相关基因c-Myc,Western blot结果证实上调ABHD11-AS1后,c-Myc表达量随之升高。结论ABHD11-AS1通过上调c-Myc促进胃癌细胞内的糖酵解,并加速胃癌进展。

Objective To explore the role of long non-coding RNAABHD11-AS1 in regulation of glycolysis in gastric cancer cells and its molecular mechanism.Methods The null plasmid pcDNA-Vector and the overexpression plasmid pcDNA-ABHD11-AS1 were transfected into human gastric cancer cell lines MKN45 and MGC803 with low ABHD11-AS1 expression,and the changes in cell proliferation,colony formation,migration and invasion were examined using CCK-8 assay,colony formation assay and Transwell assay.Glucose uptake and lactate production of the cells were detected to assess the changes in glycolytic activity.The LncMAP database was used to identify potential transcription factors regulated by ABHD11-AS1,and the candidate transcription factor was determined by literature review,and the result was verified using Western blotting.Results Transfection with pcDNA-ABHD11-AS1 significantly increased ABHD11-AS1 expression in MGC803 and MKN45 cells,which exhibited obviously accelerated cell proliferation(P<0.05),increased colony formation rate and enhanced cell migration and invasion abilities(P<0.01).ABHD11-AS1 overexpression obviously promoted glycolysis in MGC803 and MKN45 cells(P<0.05).Analysis of the database suggested that ABHD11-AS1 may regulate the classical glycolysis-related gene c-Myc in gastric cancer cells.Western blotting demonstrated that the expression of c-Myc increased significantly after upregulating ABHD11-AS1 in gastric cancer cells.Conclusion ABHD11-AS1 promotes glycolysis in gastric cancer cells by upregulating c-Myc to accelerate gastric cancer progression.