Interleukin-1 beta and transforming growth factor-alpha/epidermal growth factor induce expression of M(r) 95,000 type IV collagenase/gelatinase and interstitial fibroblast-type collagenase by rat mucosal keratinocytes.
Interleukin-1 beta and transforming growth factor-alpha/epidermal growth factor induce expression of M(r) 95,000 type IV collagenase/gelatinase and interstitial fibroblast-type collagenase by rat mucosal keratinocytes.
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DOI：
10.1016/s0021-9258(17)46745-7
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发表时间：
1993-09
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影响因子：
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通讯作者：
Lyons Jg;B. Birkedal‐Hansen;Pierson Mc;J. Whitelock;H. Birkedal-Hansen
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作者：
Lyons Jg;B. Birkedal‐Hansen;Pierson Mc;J. Whitelock;H. Birkedal-Hansen
在永久无血清条件下连续培养的大鼠粘膜角质形成细胞通过上调M(R)95,000明胶酶(MMP9)(M(R)95KGL)和成纤维细胞型胶原酶(MMP1)(FIB-CL)，对白介素1(IL)-1β/IL-α和转化生长因子(TGF)-α/表皮生长因子(EGF)(以及12-O-十四烷基佛波醇-13-乙酸酯(TPA))产生反应，而对照细胞几乎检测不到这两种酶的表达。在最大限度的诱导下，细胞分泌8-10微克/10(6)细胞/天(M(R)95K GL)和2-3微克/10(6)细胞/天(FIB-CL)的酶蛋白。到目前为止，IL-1β是最强的细胞因子，在10(-10)M时作用最大，而IL-1α、TGF-α和EGF则需要20-100倍以上的浓度。TPA(10(-7)M)可阻断细胞对IL-1β、转化生长因子-α和EGF的后续反应，同时导致胞浆蛋白激酶C活性下降90%。令人惊讶的是，星形孢子素，一种有效的激酶抑制剂，不仅没有阻止生长因子/细胞因子反应，而且本身刺激了酶的表达，其幅度与TPA相当。10(-7)M地塞米松可使转化生长因子-α/表皮生长因子的诱导作用下调70-85%。地塞米松对IL-1β反应的抑制作用较弱，M(R)95K GL减少60%，FIB-CL几乎没有减少。地塞米松也未能阻断TPA的反应。
Rat mucosal keratinocytes serially propagated under permanently serum-free conditions responded to interleukin (IL)-1 beta/IL-alpha and to transforming growth factor (TGF)-alpha/epidermal growth factor (EGF) (as well as to 12-O-tetradecanoylphorbol-13-acetate (TPA)) by upregulation of M(r) 95,000 gelatinase (MMP-9) (M(r) 95K GL) and fibroblast-type collagenase (MMP-1) (FIB-CL), whereas control cells expressed barely detectable levels of either of these enzymes. The cells secreted 8-10 micrograms/10(6) cells/day (M(r) 95K GL) and 2-3 micrograms/10(6) cells/day (FIB-CL) of enzyme protein for at least 24 h when maximally induced. This level was attained only after a 24-h lag period, and the earliest emergence of enzyme protein in the culture medium required 10-14 h. IL-1 beta was by far the most potent cytokine with maximal effect already at 10(-10) M, whereas IL-1 alpha, TGF-alpha, and EGF required 20-100-fold higher concentrations. Pretreatment of the cells with TPA (10(-7) M) abolished the subsequent response to IL-1 beta, TGF-alpha, and EGF and at the same time resulted in > 90% reduction of cytosolic protein kinase C activity. Surprisingly, staurosporine, a potent kinase inhibitor, not only failed to block growth factor/cytokine responses but itself stimulated expression of the enzymes at a magnitude comparable to TPA. The inducing effect of TGF-alpha/EGF was down-regulated by 70-85% by 10(-7) M dexamethasone. Dexamethasone was less effective in ablating the IL-1 beta response yielding 60% reduction M(r) 95K GL and little or no reduction of FIB-CL. Dexamethasone also failed to block the TPA response.