IL-15调控HaCaT细胞增殖及分化的研究
发布时间：2019-06-05 19:18
【摘要】：目的：探讨IL-15调节HaCaT细胞增殖与分化及其可能的机制。方法：培养的HaCaT细胞,不同剂量IL-15刺激不同时间后,MTT法分析IL-15对HaCaT细胞增殖的影响。IL-15处理HaCaT1d后,加入1mmol的氯化钙诱导HaCaT细胞分化,Western-blotting分析HaCaT细胞的分化Markers(颗粒层：Keratin1,基底层：Keratin5和棘层：Involucrin)观察IL-15对氯化钙诱导HaCaT细胞分化的影响。Western-blotting分析IL-15在HaCaT细胞中激活的信号通路：MAPKs-ERK1/2和PI3K-AKT。在IL-15处理HaCaT细胞前1h,分别加入MAPKs-ERK1/2和PI3K-AKT特异性抑制PD98059和LY2940002以阻断IL-15激活的相关信号通路,分析阻断剂对IL-15调控HaCaT细胞增殖及分化影响,探讨这些信号通路在IL-15对HaCaT细胞增殖及分化调控中的作用。结果：MTT分析结果显示：IL-15显著促进HaCaT细胞增殖,且具有时间及剂量依赖性；IL-15能够抑制氯化钙诱导的HaCaT细胞分化markers (Keratin1及Involucrin)的表达,存在时间效应及剂量效应；信号通路分析揭示IL-15能够增加pERKl/2及pAKT的水平；阻断剂的研究发现IL-15部分依赖Ras-MAPKs(ERK1/2)和PI3K-PAKT途径发挥其促进增殖及抑制分化作用。结论：IL-15部分依赖Ras-MAPKs-PERK1/2和PI3K-PAKT信号途径促进HaCaT细胞增殖和抑制分化。
[Abstract]:Objective: to investigate the effect of IL-15 on the proliferation and differentiation of HaCaT cells and its possible mechanism. Methods: cultured HaCaT cells were stimulated with different doses of IL-15 for different time. The effect of IL-15 on the proliferation of HaCaT cells was analyzed by MTT assay. After HaCaT1d was treated with IL-15, HaCaT cells were induced to differentiate by adding 1mmol calcium chloride. Western-blotting Analysis of differentiated Markers (granule layer: Keratin1,) in HaCaT cells Basal layer: Keratin5 and spinous layer: Involucrin) were used to observe the effect of IL-15 on the differentiation of HaCaT cells induced by calcium chloride. Western-blotting analysis of the signal pathways activated by IL-15 in HaCaT cells: MAPKs-ERK1/2 and PI3K-AKT. One hour before HaCaT cells were treated with IL-15, MAPKs-ERK1/2 and PI3K-AKT were added to specifically inhibit PD98059 and LY2940002 to block the signal pathway related to IL-15 activation, and the effects of blockers on the proliferation and differentiation of HaCaT cells induced by IL-15 were analyzed. To investigate the role of these signaling pathways in the regulation of IL-15 on the proliferation and differentiation of HaCaT cells. Results: MTT analysis showed that IL-15 significantly promoted the proliferation of HaCaT cells in a time-and dose-dependent manner, and IL-15 could inhibit the expression of markers (Keratin1 and Involucrin) induced by calcium chloride in HaCaT cells, and there was a time effect and dose effect. Signal pathway analysis showed that IL-15 could increase the levels of pERKl/2 and pAKT, and the study of blockers found that IL-15 partly relied on Ras-MAPKs (ERK1/2) and PI3K-PAKT pathway to promote proliferation and inhibit differentiation. Conclusion: IL-15 partly relies on Ras-MAPKs-PERK1/2 and PI3K-PAKT signaling pathways to promote the proliferation and inhibit differentiation of HaCaT cells.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
本文编号：2493764
[Abstract]:Objective: to investigate the effect of IL-15 on the proliferation and differentiation of HaCaT cells and its possible mechanism. Methods: cultured HaCaT cells were stimulated with different doses of IL-15 for different time. The effect of IL-15 on the proliferation of HaCaT cells was analyzed by MTT assay. After HaCaT1d was treated with IL-15, HaCaT cells were induced to differentiate by adding 1mmol calcium chloride. Western-blotting Analysis of differentiated Markers (granule layer: Keratin1,) in HaCaT cells Basal layer: Keratin5 and spinous layer: Involucrin) were used to observe the effect of IL-15 on the differentiation of HaCaT cells induced by calcium chloride. Western-blotting analysis of the signal pathways activated by IL-15 in HaCaT cells: MAPKs-ERK1/2 and PI3K-AKT. One hour before HaCaT cells were treated with IL-15, MAPKs-ERK1/2 and PI3K-AKT were added to specifically inhibit PD98059 and LY2940002 to block the signal pathway related to IL-15 activation, and the effects of blockers on the proliferation and differentiation of HaCaT cells induced by IL-15 were analyzed. To investigate the role of these signaling pathways in the regulation of IL-15 on the proliferation and differentiation of HaCaT cells. Results: MTT analysis showed that IL-15 significantly promoted the proliferation of HaCaT cells in a time-and dose-dependent manner, and IL-15 could inhibit the expression of markers (Keratin1 and Involucrin) induced by calcium chloride in HaCaT cells, and there was a time effect and dose effect. Signal pathway analysis showed that IL-15 could increase the levels of pERKl/2 and pAKT, and the study of blockers found that IL-15 partly relied on Ras-MAPKs (ERK1/2) and PI3K-PAKT pathway to promote proliferation and inhibit differentiation. Conclusion: IL-15 partly relies on Ras-MAPKs-PERK1/2 and PI3K-PAKT signaling pathways to promote the proliferation and inhibit differentiation of HaCaT cells.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
【参考文献】
相关期刊论文 前3条
1 李惠丽;陈铭珍;;白细胞介素-15与临床疾病相关性研究进展[J];实用儿科临床杂志;2007年09期
2 陈铭珍,叶中绿,蔡康荣,黄秀兰,陈日玲,程涵蓉;IL-15对MDS患者CD34~+细胞诱导增殖分化和抗凋亡作用[J];中国实验血液学杂志;2005年04期
3 梁淑娟,孙lm,魏海明,孙安源,张建华,田志刚;IL-15通过ERK1/2信号途径对NK细胞活性的调节[J];中华微生物学和免疫学杂志;2003年02期
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