长链非编码RNA MALAT1对槟榔碱诱导的颊黏膜成纤维细胞活化能力影响的研究

LncRNA MALAT1 Inhibits Buccal Mucosal Fibroblasts Activities in Oral Submucous Fibrosis

目的:本研究旨在明确长链非编码RNA(long non-coding RNA,lncRNA) MALAT1对槟榔碱诱导的颊黏膜成纤维细胞(buccal mucosal fibroblasts, BMFs)活化能力影响的作用,为寻找新的口腔黏膜下纤维化(oral submucous fibrosis, OSF)治疗途径提供理论基础。方法:体外培养BMFs,采用不同浓度槟榔碱刺激BMFs,在细胞水平上构建OSF疾病模型。采用细胞计数试剂盒(cell counting kit-8,CCK-8)法检测细胞活力;Transwell法和胶原凝胶收缩法检测BMFs活化;实时荧光定量聚合酶链反应(quantitative real-time PCR, qRT-PCR)检测BMFs中平滑肌肌动蛋白-α(α-smooth muscle actin, α-SMA)和lncRNA MALAT1的表达情况;siRNA-MALAT1瞬时转染到BMFs中,评估槟榔碱刺激下BMFs迁移和收缩能力的变化。结果:10μg/mL槟榔碱是体外构建OSF疾病模型的最佳浓度,此时细胞内α-SMA和lncRNA MALAT1表达升高。Transwell实验和胶原凝胶收缩实验提示敲低lncRNA MALAT能抑制槟榔碱诱导的BMFs收缩和迁移。结论:lncRNA MALAT1对槟榔碱诱导的BMFs激活,细胞收缩和迁移有重要作用。

Objective:To clarify the effect of long non-coding RNA MALAT1 on the activation ability of buccal mucosal fibroblasts(BMFs)induced by arecoline,and to establish theoretical basis for a new therapeutic pathway to oral submucous fibrosis(OSF).Methods:BMFs were cultured in vitro,and were stimulated with different concentrations of arecoline to construct OSF-disease models at the cellular level.CCK-8 method was used to detect cell viability.BMFs activation was detected by Transwell method and collagen gel shrinkage method.Quantitative real-time PCR(qRT-PCR)was used to detect the expression ofα-smooth muscle actin antibody(α-SMA)and lncRNA MALAT1 in BMFs.siRNA-MALAT1 was transiently transfected into BMFs to assess changes in the migration and contraction capacity of BMFs stimulated by arecoline.Results:10μg/mL of arecoline was the optimal concentration for constructing OSF-disease models in vitro,when intracellular expression ofα-SMA and lncRNA MALAT1 was elevated.Transwell experiments and collagen gel contraction experiments suggested that knockdown lncRNA MALAT inhibited arecoline-induced BMFs contraction and migration.Conclusion:lncRNA MALAT1 plays an important role in arecoline-induced BMFs activation,cell contraction,and migration.