长链非编码RNA MALAT1对人肺成纤维细胞向肌成纤维细胞转化的影响

常清; 陈飞; 张铁栓; 乔华; 万军营

doi:10.12019/j.issn.1671-5144.2023.02.008

长链非编码RNA MALAT1对人肺成纤维细胞向肌成纤维细胞转化的影响

Effects of Long Non-Coding RNA MALAT1 on the Transformation of Human Lung Fibroblasts Into Myofibroblasts

摘要

摘要:
目的探索长链非编码RNA（long non-coding RNA，lncRNA）肺腺癌转移相关转录本1（metastasis-associated lung adenocarcinoma transcript 1，MALAT1）对人肺成纤维细胞向肌成纤维细胞转化的影响。
方法培养人肺成纤维细胞（human fetal lung fibroblast 1，HFL-1），按照有无转化生长因子-β1（transforming growth factor-β1，TGF-β1）作用分为观察组和对照组，检测两组细胞中lncRNA MALAT1表达水平。在此基础上设置四个组别：空白组、TGF-β1组、转染对照组和siMALAT1组。空白组细胞不接受任何处理；TGF-β1组采用5 ug/L的TGF-β1处理；转染对照组在TGF-β1组的基础上应用siMALAT1阴性对照（negative control，NC）处理；siMALAT1组在TGF-β1组的基础上应用siMALAT1。比较其lncRNA MALAT1、Ⅰ型胶原（collagen 1，COL1）和α-平滑肌肌动蛋白（α-smooth muscle activation protein，α-SMA）的mRNA表达水平，COL1和α-SMA的蛋白表达水平和细胞增殖情况。
结果观察组lncRNA MALAT1相对表达量较对照组高（P<0.05）。四组细胞lncRNA MALAT1、α-SMA和COL1的mRNA及蛋白相对表达量有统计学差异（P<0.05）；其中，转染对照组和TGF-β1组以上RNA和蛋白相对表达量均高于siMALAT1组和空白组（P<0.05），而TGF-β1组与转染对照组之间上述RNA和蛋白相对表达量则无差异（P>0.05）。四组细胞增殖活力的差异具有统计学意义（P<0.05）；转染对照组和TGF-β1组细胞增殖活力较siMALAT1组和空白组高（P<0.05），而TGF-β1组与转染对照组之间则无差异（P>0.05）。
结论 lncRNA MALAT1可以促进人肺成纤维细胞向肌成纤维细胞的转化，影响肺纤维化进程中COL1、α-SMA在肺组织中的沉积，靶向lncRNA MALAT1将有助于对肺纤维化过程的深入研究。

Abstract:
Objective To explore the effect of long noncoding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) on the transformation of human lung fibroblasts into myofibroblasts.
Methods The cultured human lung fibroblasts 1 (HFL-1) were divided into an observation group and a control group according to whether they have the effect of transforming growth factor-β1 (TGF-β1). Detected the expression levels of lncRNA MALAT1 in cells of the two groups. On this basis, four clusters were established: blank group, TGF-β1 group, transfection control group, and siMALAT1 group. Cells in the blank group did not receive any treatment; the TGF-β1 group was treated with TGF-β1 at 5 μg/L; the transfection control group was treated with siMALAT1 negative control (NC) based on the TGF-β1 group; siMALAT1 group was treated with siMALAT1 based on the TGF-β1 group. The lncRNA MALAT1, mRNA expression levels of collagen 1 (COL1) and α-smooth muscle activation protein (α-SMA), protein expression levels of COL1 and α-SMA, and cell proliferation were compared among them.
Results Compared with the control group, lncRNA MALAT1 relative expression in the observation group was higher (P<0.05). There were statistically significant differences in the four groups on lncRNA MALAT1, mRNA expression levels of COL1 and α-SMA, and protein expression levels of COL1 and α-SMA (P<0.05). The relative expression of RNA and protein above in the transfection control group and TGF-β1 group was higher than that in the siMALAT1 group and blank group (P<0.05), but there were no significant between the TGF-β1 group and the transfection control group (P>0.05). The four groups showed significant differences in cell viability (P<0.05). The proliferation activity of the transfection control group and TGF-β1 group was higher than that of the siMALAT1 group and blank group (P<0.05), however, there were no significant between the TGF-β1 group and the transfection control group (P>0.05).
Conclusions LncRNA MALAT1 can promote the transformation of human lung fibroblasts into myofibroblasts, affecting the deposition of COL1, α-SMA in the lung tissue during the course of pulmonary fibrosis, and targeting lncRNA MALAT1 would contribute to the in-depth study of the process of pulmonary fibrosis.

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