电离辐射对瘢痕细胞生物学行为及表达谱的影响
首发时间:2021-02-04
摘要:目的:瘢痕疙瘩(Keloid, KD)是成纤维细胞过度增殖以及胶原蛋白过度沉积为特征的真皮纤维化疾病。手术切除联合放射治疗是目前治疗瘢痕疙瘩最有效的方法,但瘢痕组织存在的辐射抵抗仍然是限制其治疗效果不可忽略的因素。因此,探究瘢痕疙瘩的辐射抵抗及其增敏机制显得极为重要。方法:采用组织块培养法培养瘢痕疙瘩原代细胞;采用EDU和细胞克隆形成实验检测细胞的增殖能力;通过检测β-半乳糖苷酶染色检测细胞衰老;采用Western blot检测蛋白质表达水平;通过对受照细胞进行mRNA基因组测序,筛选差异表达基因以及通过生物信息学相关软件进行转录因子的预测。结果:瘢痕疙瘩原代细胞多数呈长梭形,细胞质向外伸出两三个长短不同的突起;X 射线能明显的抑制瘢痕疙瘩细胞的增殖以及促进细胞的衰老;在4 Gy和8 Gy照射后的瘢痕原代细胞中,分别有184个和204个基因发生了改变;上调基因富集到7条具有显著差异的通路,包括细胞衰老、FOXO通路等;下调基因富集到5条信号通路,包括核糖体、氨基酸的生物合成等信号通路。通过生物信息学预测到TIN、ESR1以及PPARγ等转录因子参与上述表达谱变化。结论:该研究揭示了电离辐射照射瘢痕原代细胞后的细胞表型及mRNA变化,并为瘢痕辐射抵抗提供潜在的增敏靶点。
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Effects of ionizing radiation on the biological behavior and expression profile of keloid
Abstract:Objective: Keloid is a dermal fibroproliferative disease characterized by abnormal proliferation of fibroblasts and massive accumulation of extracellular matrix. Surgical excision followed by radiotherapy is considered as a reliable treatment for keloid, however, radioresistance remains a serious impediment to treatment efficacy. Therefore, it is extremely important to explore the radiation resistance and sensitization mechanism of keloids. Methods: Tissue block culture was used to culture primary keloid fibroblasts; EDU and clonogenic assay were used to detect cell proliferation; SA β-gal staining and Western blot were used to observe the effects of ionizing radiation on cell senescence. By sequencing the mRNA genome of ionizing cells, screening differentially expressed genes, and predicting transcription factors through bioinformatics-related software. Results: The primary cells of keloids were mostly long spindle-shaped, and the cytoplasm protruded out of two or three protrusions of different lengths; X-rays could significantly inhibit the proliferation and promote senescence of keloid cells;184 and 204 genes were changed respectively in 4 and 8 Gy; Up-regulated gene enrichment to 7 significant pathways, including Cellular senescence, FOXO pathway; Down-regulated gene enrichment to 5 signaling pathways, including Ribosome, Biosynthesis of amino acids and other signaling pathways. It is predicted by bioinformatics that transcription factors such as TIN, ESR1 and PPARG participate in the above expression profile changes. Conclusion: This study reveals the cell phenotype and mRNA changes of primary scar cells after ionizing radiation exposure, and provides a potential sensitizing target for scar radiation resistance.
Keywords: keloid, ionizing radiation, expression profile sequencing
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电离辐射对瘢痕细胞生物学行为及表达谱的影响
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