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| 下载次数 | 被引频次 | 阅读次数 |
目的：研究香烟模拟物（cigarette smoke extract, CES）通过调控16 HBE支气管上皮细胞中外泌体miR-34a的表达，从而影响MRC-5肺成纤维细胞的增殖。方法：从市售香烟中制备CES，后将6HBE细胞经过CES处理，收集上清中的外泌体miR-34a用于MRC-5细胞培养；使用RT-PCR检测外泌体miR-34a的表达水平；MRC-5细胞增殖能力通过细胞计数试剂盒CCK-8测定；通过Western blot检测CASP2的表达，利用双荧光素酶验证miR-34a与CASP2基因的靶向结合。结果：透射电镜下，16 HBE上清液中的外泌体呈球状，粒径在100 nm左右；经过CES处理后，外泌体miR-34a的表达显著增加。进一步研究表明，通过CES诱导的外泌体miR-34a能够促进MRC-5细胞的增殖；miR-34a与CASP2存在靶向结合关系；miR-34a mimic显著抑制了CASP2的表达。结论：在CES诱导的16HBE细胞中，外泌体miR-34a通过与CASP2基因的靶向结合，对成纤维细胞增殖起着关键作用。
Abstract：Objective: To explore how cigarette smoke extract(CES) regulates the expression of exosomal miR-34a in 16HBE bronchial epithelial cells, thus affecting the proliferation of MRC-5 lung fibroblasts. Methods: CES was prepared from commercially available cigarettes, and 16HBE cells were treated with CES. The exosomal miR-34a collected from the supernatant was used for MRC-5 cell culture. The expression level of exosomal miR-34a was detected by RT-PCR. The proliferation ability of MRC-5 cells was determined by CCK-8 cell counting kit. The expression of CASP2 was detected by Western blot, and the target binding of miR-34a and CASP2 gene was verified by dual luciferase.Results: Under the transmission electron microscope, the exosomes in the supernatant of 16 HBE were spherical, with a particle size of about 100 nm; after CES treatment, the expression of exosomal miR-34a significantly increased. Further research showed that the exosomal miR-34a induced by CES can promote the proliferation of MRC-5 cells; miR-34a and CASP2 have a target binding relationship; miR-34a mimic significantly inhibited the expression of CASP2. Conclusion: In CES-induced 16HBE cells, exosomal miR-34a plays a key role in fibroblast proliferation through target binding with the CASP2 gene.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20230718.001
中图分类号:R563.9
引用信息:
[1]李思广,林凌桑,陈洁,等.香烟诱导支气管上皮细胞外泌体释放miR-34a靶向CASP2促进肺成纤维细胞增殖[J].海南医学院学报,2023,29(17):1288-1294+1301.DOI:10.13210/j.cnki.jhmu.20230718.001.
基金信息:
海南省自然科学基金青年基金（822QN447）; 国家自然科学基金资助（82160011）~~
2023-07-18
2023-07-18
2023-07-18