Zhu Zhankun, Dai Jinhua, Liao Yufeng, Wang Tao
Ningbo No. 2 Hospital, Department of Clinical Laboratory, Ningbo, Zhejiang, 315010, China.
Biochemistry (Mosc). 2017 May;82(5):606-612. doi: 10.1134/S000629791705008X.
Sex-determining region Y-box 9 (Sox9) is an important transcription factor that has been identified as a key regulator of several types of diseases. In this study, we explored the correlation of Sox9 with cell proliferation, apoptosis, inflammatory factor expression, and the possible signaling pathway in human lung fibroblast cell line to investigate the possible mechanism of neonatal pneumonia. Therefore, in the present study, pc-Sox9 and si-Sox9 were transfected into MRC-5 (human fetal lung fibroblast cell line) to promote or inhibit expression of Sox-9. Quantitative reverse-transcription polymerase chain reaction and Western blot were used to determine the expression level of Sox-9 at mRNA and protein level. Then 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and flow cytometry were used to explore, respectively, proliferation and apoptosis in vitro. We found that Sox9 could significantly upregulate the proliferation rate and inhibit apoptosis rate and inflammatory factor expression of MRC-5 cells compared with a control group. Moreover, the signaling pathway study confirmed that Sox9 protected MRC-5 from lipopolysaccharide injury through the AKT/GSK3β pathway. All these findings suggest that Sox9 acts as a novel marker for neonatal pneumonia and could be a new therapeutic target for this disease.
性别决定区Y框蛋白9（Sox9）是一种重要的转录因子，已被确定为多种疾病的关键调节因子。在本研究中，我们探讨了Sox9与细胞增殖、凋亡、炎性因子表达的相关性，以及其在人肺成纤维细胞系中的可能信号通路，以研究新生儿肺炎的可能机制。因此，在本研究中，将pc-Sox9和si-Sox9转染至MRC-5（人胎儿肺成纤维细胞系）中，以促进或抑制Sox-9的表达。采用定量逆转录聚合酶链反应和蛋白质免疫印迹法分别在mRNA和蛋白质水平测定Sox-9的表达水平。然后分别采用噻唑蓝（MTT）比色法和流式细胞术检测体外细胞增殖和凋亡情况。我们发现，与对照组相比，Sox9可显著提高MRC-5细胞的增殖率，抑制其凋亡率和炎性因子表达。此外，信号通路研究证实，Sox9通过AKT/GSK3β信号通路保护MRC-5细胞免受脂多糖损伤。所有这些发现表明，Sox9可作为新生儿肺炎的新型标志物，可能成为该病的新治疗靶点。
