【摘要】：目的：探討IL-15調(diào)節(jié)HaCaT細(xì)胞增殖與分化及其可能的機(jī)制。方法：培養(yǎng)的HaCaT細(xì)胞,不同劑量IL-15刺激不同時間后,MTT法分析IL-15對HaCaT細(xì)胞增殖的影響。IL-15處理HaCaT1d后,加入1mmol的氯化鈣誘導(dǎo)HaCaT細(xì)胞分化,Western-blotting分析HaCaT細(xì)胞的分化Markers(顆粒層：Keratin1,基底層：Keratin5和棘層：Involucrin)觀察IL-15對氯化鈣誘導(dǎo)HaCaT細(xì)胞分化的影響。Western-blotting分析IL-15在HaCaT細(xì)胞中激活的信號通路：MAPKs-ERK1/2和PI3K-AKT。在IL-15處理HaCaT細(xì)胞前1h,分別加入MAPKs-ERK1/2和PI3K-AKT特異性抑制PD98059和LY2940002以阻斷IL-15激活的相關(guān)信號通路,分析阻斷劑對IL-15調(diào)控HaCaT細(xì)胞增殖及分化影響,探討這些信號通路在IL-15對HaCaT細(xì)胞增殖及分化調(diào)控中的作用。結(jié)果：MTT分析結(jié)果顯示：IL-15顯著促進(jìn)HaCaT細(xì)胞增殖,且具有時間及劑量依賴性；IL-15能夠抑制氯化鈣誘導(dǎo)的HaCaT細(xì)胞分化markers (Keratin1及Involucrin)的表達(dá),存在時間效應(yīng)及劑量效應(yīng)；信號通路分析揭示IL-15能夠增加pERKl/2及pAKT的水平；阻斷劑的研究發(fā)現(xiàn)IL-15部分依賴Ras-MAPKs(ERK1/2)和PI3K-PAKT途徑發(fā)揮其促進(jìn)增殖及抑制分化作用。結(jié)論：IL-15部分依賴Ras-MAPKs-PERK1/2和PI3K-PAKT信號途徑促進(jìn)HaCaT細(xì)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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R363

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