【摘要】 目的探讨lncRNA LOXL1-AS1对胃癌细胞侵袭、迁移的影响及其作用机制。方法 Transwell小室法检测上调或敲低LOXL1-AS1表达后对胃癌细胞侵袭、迁移能力的影响,生物信息学方法预测LOXL1-AS1和miR-142-5p的靶向关系,双荧光素酶报告基因实验验证两者靶向关系,实时荧光定量PCR(qRT-PCR)法检测上调或敲低LOXL1-AS1表达对miR-142-5p表达的影响;免疫印迹法检测EMT标志物蛋白(E-cadherin、N-cadherin、Vimentin、ZO-1)表达和PIK3CA表达。结果敲低LOXL1-AS1表达可抑制胃癌细胞侵袭、迁移能力,并抑制胃癌细胞EMT;双荧光素酶实验证实LOXL1-AS1直接靶向作用于miR-142-5p;LOXL1-AS1负调控miR-142-5p表达与活性,并通过miR-142-5p正调控PIK3CA表达;过表达LOXL1-AS1可促进胃癌细胞侵袭和迁移,miR-142-5p可逆转LOXL1-AS1过表达对胃癌细胞侵袭和迁移的促进作用。结论 LOXL1-AS1靶向miR-142-5p/PIK3CA调控胃癌细胞侵袭和迁移能力。更多还原

【Abstract】 Objective To investigate the effect of lncRNA LOXL1-AS1 on invasion and migration of gastric cancer cells, and explore the potential mechanisms. Methods Transwell assay was conducted to detect the effect of LOXL1-AS1 on invasion and migration of gastric cancer cells. The targeted relationship of miR-142-5 p and LOXL1-AS1 was predicted by bioinformatics analysis, and Luciferase reporter assay was used to confirm targeting of miR-142-5 p by LOXL1-AS1. Quantitive realtime PCR(qRT-PCR) was used to detect the effect of LOXL1-AS1 on the expression of miR-142-5 p. The expressions of E-cadherin, N-cadherin, Vimentin, ZO-1 and PIK3 CA were evaluated by Western blotting. Results Knockdown LOXL1-AS1 could inhibit invasion, migration and EMT in gastric cancer cells. The targeted relationship of miR-142-5 p and LOXL1-AS1 was confirmed by Luciferase reporter assay. The expression of miR-142-5 p was decreased in LOXL1-AS1 overexpressed cells whereas the expression of miR-142-5 p was increased in cells with LOXL1-AS1 knockdown. LOXL1-AS1 could modulate the expression of PIK3 CA in gastric cancer cells via miR-142-5 p. The overexpression of LOXL1-AS1 significantly facilitated the invasion and migration of gastric cancer cells, and miR-142-5 p could reverse the effect induced by overexpressed LOXL1-AS1 on the invasion and migration of gastric cancer cells. Conclusion LOXL1-AS1 can promote invasion and migration of gastric cancer cells by targeting miR-142-5 p/PIK3 CA axis.更多还原