胰岛素样生长因子结合蛋白7通过调控TGF-β1信号通路抑制HaCaT细胞生长
发布时间：2019-02-12 19:53
【摘要】：背景胰岛素样生长因子结合蛋白7(IGFBP7)被发现作为为数不多的角质形成细胞特异基因中的一种。一项全基因表达研究显示IGFBP7在银屑病表皮中的表达下调。前期的研究发现,IGFBP7下调对转化生长因子β(TGF-β)有影响,但是具体机制不明确。目的为了全面评估IGFBP7在银屑病调节中的作用,以及其与TGF-β通路复杂效应的分子机制,我们检测了IGFBP7与转化生长因子β1(TGF-β1)和其下游p38 MAPK的活性之间的联系。 方法HaCaT细胞是一种自主永生化的人类角质形成细胞系。我们通过在HaCaT细胞中使用IGFBP7特异性小干扰RNA (IGFBP7- siRNA)或重组IGFBP7 (rIGFBP7)来人为地下调或上调IGFBP7的水平。利用细胞免疫组化方法检测IGFBP7和TGF-β1的表达水平。MTT法和annexin V/PI法用来检测细胞活性和细胞凋亡的水平。蛋白印迹法用来检测IGFBP7、TGF-β1和p38-MAPK的蛋白水平。 结果我们的研究发现IGFBP7下调显著提高了HaCaT细胞的增殖能力,其与细胞的凋亡水平明显降低有关。而重组IGFBP7可以逆转上述效应。IGFBP7沉默的细胞中,TGF-β1和磷酸化MAPK水平上调,而IGFBP7过表达细胞中TGF-β1和磷酸化MAPK水平下调。 结论我们的结果提示IGFBP7在HaCaT细胞中对TGF-β1的调控可能通过MAPK通路,这为增厚性皮肤病的治疗提供了线索。
[Abstract]:Background Insulin-like growth factor binding protein 7 (IGFBP7) has been identified as one of the few keratinocyte specific genes. A whole gene expression study showed that IGFBP7 expression was down-regulated in psoriatic epidermis. Previous studies have found that down-regulation of IGFBP7 has an effect on transforming growth factor 尾 (TGF- 尾), but the mechanism is unclear. Objective to evaluate the role of IGFBP7 in psoriasis regulation and the molecular mechanism of its complex effect on TGF- 尾 pathway, we examined the relationship between IGFBP7 and the activity of transforming growth factor 尾 1 (TGF- 尾 1) and its downstream p38 MAPK. Methods HaCaT cell line is an autonomous immortalized human keratinocyte cell line. We artificially down-regulated or upregulated IGFBP7 levels by using IGFBP7 specific small interfering RNA (IGFBP7- siRNA) or recombinant IGFBP7 (rIGFBP7) in HaCaT cells. The expression of IGFBP7 and TGF- 尾 1 was detected by immunohistochemistry, and the activity and apoptosis of cells were detected by MTT and annexin V/PI methods. Protein levels of IGFBP7,TGF- 尾 1 and p38-MAPK were detected by Western blot. Results in our study, down-regulation of IGFBP7 significantly increased the proliferation of HaCaT cells, which was related to the decrease of apoptosis level. The recombinant IGFBP7 could reverse the above effects. The levels of TGF- 尾 1 and phosphorylated MAPK were up-regulated in IGFBP7 silencing cells, while TGF- 尾 1 and phosphorylated MAPK levels were down-regulated in IGFBP7 overexpression cells. Conclusion our results suggest that the regulation of TGF- 尾 1 by IGFBP7 in HaCaT cells may be mediated by MAPK pathway, which may provide clues for the treatment of thickening dermatosis.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R329
本文编号：2420735
[Abstract]:Background Insulin-like growth factor binding protein 7 (IGFBP7) has been identified as one of the few keratinocyte specific genes. A whole gene expression study showed that IGFBP7 expression was down-regulated in psoriatic epidermis. Previous studies have found that down-regulation of IGFBP7 has an effect on transforming growth factor 尾 (TGF- 尾), but the mechanism is unclear. Objective to evaluate the role of IGFBP7 in psoriasis regulation and the molecular mechanism of its complex effect on TGF- 尾 pathway, we examined the relationship between IGFBP7 and the activity of transforming growth factor 尾 1 (TGF- 尾 1) and its downstream p38 MAPK. Methods HaCaT cell line is an autonomous immortalized human keratinocyte cell line. We artificially down-regulated or upregulated IGFBP7 levels by using IGFBP7 specific small interfering RNA (IGFBP7- siRNA) or recombinant IGFBP7 (rIGFBP7) in HaCaT cells. The expression of IGFBP7 and TGF- 尾 1 was detected by immunohistochemistry, and the activity and apoptosis of cells were detected by MTT and annexin V/PI methods. Protein levels of IGFBP7,TGF- 尾 1 and p38-MAPK were detected by Western blot. Results in our study, down-regulation of IGFBP7 significantly increased the proliferation of HaCaT cells, which was related to the decrease of apoptosis level. The recombinant IGFBP7 could reverse the above effects. The levels of TGF- 尾 1 and phosphorylated MAPK were up-regulated in IGFBP7 silencing cells, while TGF- 尾 1 and phosphorylated MAPK levels were down-regulated in IGFBP7 overexpression cells. Conclusion our results suggest that the regulation of TGF- 尾 1 by IGFBP7 in HaCaT cells may be mediated by MAPK pathway, which may provide clues for the treatment of thickening dermatosis.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R329
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