长链非编码RNA-PVT1介导JAK/STAT3信号通路对肺纤维化的研究

目的 探讨长链非编码RNA-浆细胞瘤变异易位基因1/蛋白质酪氨酸激酶/信号转导和转录激活因子3（lnc-PVT1/JAK/STAT3）信号通路参与肺纤维化的作用机制。方法 采用不同浓度脂多糖（LPS）作用于人正常肺上皮细胞BEAS-2B，用CCK-8法检测细胞活性、蛋白免疫印迹法检测α-SMA的蛋白相对表达量，对LPS...

Long non-coding RNA-PVT1 induces the progression of pulmonary fibrosis via the JAK/STAT3 signaling pathway

Objective To investigate the role of long non-coding RNA-PVT1/JAK/STAT3 signaling pathway in pulmonary fibrosis. Methods BEAS-2B cells were treated with different concentrations of LPS. Cell viability was assessed by CCK-8. The relative expression level of α-SMA protein was determined by western blot. The concentration of LPS was screened. BEAS-2B cells were treated with the optimal concentration of LPS to establish the pulmonary fibrosis model in vitro. BEAS-2B cells were transiently transfected with lnc-PVT1 siRNA and control siRNA followed by LPS treatment. The expression level of lnc-PVT1 was determined by RT-qPCR. Effective inhibitory targets were selected for subsequent experiment. BEAS-2B cells were divided into the control （Con）group, LPS group, LPS+siRNA control （LPS+siNC） group and LPS+siPVT1 group. After transiently transfected with siPVT1 and siRNA control, BEAS-2B cells were treated with 10μg/mL LPS for 24 h. Cell viability was assessed by CCK-8. Cell migration was evaluated by cell scratch test. The relative expression level of lnc-PVT1 mRNA was measured by RT-qPCR. The expression levels of IL-6,IL-1β and TNF-α in the cultured supernatant were measured by ELISA. The expression levels of α-SMA, STAT3 and p-STAT3 were assessed by western blot. Results In the LPS group treated with 10 μg/mL LPS for 24 h, the cell proliferation was significantly higher and the expression level of fibrotic marker α-SMA protein was remarkably up-regulated than those in the Con group （both P < 0.01）. Compared with the LPS group, knockdown of lnc-PVT1 could significantly inhibit the cell proliferation and migration induced by LPS （both P < 0.01）, down-regulate the expression levels of IL-6, IL-1β and TNF-α （all P < 0.01）, and decrease the expression levels of α-SMA mRNA and protein as well as the phosphorylated STAT3 protein （all P < 0.01）. However, the relative expression level of total STAT3 protein was not affected. Conclusion Knockdown of lnc-PVT1 can effectively attenuate the LPS-induced BEAS-2B cell fibrosis, which may be associated with the role of lnc-PVT1 in regulating the JAK/STAT3 signaling pathway.