Suppression of tunicamycin-induced CD44v6 ectodomain shedding and apoptosis is correlated with temporal expression patterns of active ADAM10, MMP-9 and MMP-13 proteins in Caki-2 renal carcinoma cells
Suppression of tunicamycin-induced CD44v6 ectodomain shedding and apoptosis is correlated with temporal expression patterns of active ADAM10, MMP-9 and MMP-13 proteins in Caki-2 renal carcinoma cells
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DOI：
10.3892/or.2012.1986
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发表时间：
2012-11-01
期刊：
影响因子：
4.2
通讯作者：
Jung, Jae-Chang
中科院分区：
文献类型：
--
作者：
Kim, Yeoun-Hee;Jung, Jae-Chang
CD 44 v6已显示出协调抗凋亡分子的活化以及对凋亡的抗性。在此，我们研究了CD 44 v6胞外区在Caki-2人肾癌细胞中的脱落及其潜在机制。细胞暴露于衣霉素（TM）诱导的凋亡伴随着半胱天冬酶-3、PARP-1和CD 44 v6胞外域的裂解。TM诱导的细胞凋亡也与内质网（ER）应激密切相关，如GRP-78和CHOP蛋白表达增加所示。此外，诱导基质金属蛋白酶（MMP）-13，MMP-9和ADAM 10的表达高度刺激衣霉素的时间和剂量依赖性的方式。TM诱导的PARP-1裂解被GM 6001（一种广谱MMP抑制剂）、MMP-9/-13抑制剂和GI 254023 X（特异性ADAM 10抑制剂）显著抑制。此外，抑制所有检查MMP导致逆转TM诱导的细胞凋亡以及增加细胞活力。当考虑MMP-9和ADAM 10的功能意义时，可能活性MMP-9和ADAM 10在TM诱导的Caki-2细胞凋亡期间通过切割CD 44 v6胞外域来帮助调节CD 44 v6的细胞水平。总的来说，这些发现表明多种TM诱导的MMPs可能协同诱导细胞凋亡。
CD44v6 has been shown to coordinate the activation of anti-apoptotic molecules as well as resistance to apoptosis. Here, we investigated CD44v6 ectodomain shedding in Caki-2 human renal carcinoma cells as well as its underlying mechanisms. Exposure of cells to tunicamycin (TM)-induced apoptosis was accompanied by cleavage of caspase-3, PARP-1 and CD44v6 ectodomain. TM-induced apoptosis was also closely associated with endoplasmic reticulum (ER) stress, as shown by increased expression of GRP-78 and CHOP proteins. Furthermore, induction of matrix metalloproteinase (MMP)-13, MMP-9 and ADAM10 expression was highly stimulated by tunicamycin in a time- and dose-dependent manner. TM-induced PARP-1 cleavage was significantly inhibited by treatment with GM6001 (a broad spectrum MMP inhibitor), MMP-9/-13 inhibitor and GI254023X (specific ADAM10 inhibitor). In addition, inhibition of all examined MMPs resulted in reversal of TM-induced apoptosis as well as increased cell viability. When considering the functional implications of MMP-9 and ADAM 10, it is likely that active MMP-9 and ADAM10 help regulate the cellular levels of CD44v6 through cleavage of CD44v6 ectodomain during TM-induced apoptosis of Caki-2 cells. Collectively, these findings suggest that multiple TM-induced MMPs may cooperate to induce apoptosis.