Role of calpain-9 and PKC-delta in the apoptotic mechanism of lumen formation in CEACAM1 transfected breast epithelial cells.
Role of calpain-9 and PKC-delta in the apoptotic mechanism of lumen formation in CEACAM1 transfected breast epithelial cells.
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DOI：
10.1016/j.yexcr.2009.11.001
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发表时间：
2010-02-15
影响因子：
3.7
通讯作者：
Shively JE
中科院分区：
文献类型：
--
作者：
Chen CJ;Nguyen T;Shively JE
CEACAM 1 -4S（癌胚抗原相关细胞粘附分子1）是具有短（12个氨基酸）胞质尾的I型膜蛋白。野生型CEACAM 1 -4S转染的MCF 7细胞在3D培养中生长时形成具有管腔的腺体，而胞质结构域中两个推定的磷酸化位点（T457 A和S459 A）的无效突变未能经历管腔形成。当对从在3D培养物中生长的野生型和T457 A、S459 A突变的CEACAM 1 -4S转染的MCF 7细胞分离的mRNA进行基因芯片分析时，从超过400个基因中鉴定出钙蛋白酶-9（CAPN 9），其具有> 2log 2的差异作为管腔形成的潜在诱导剂。通过RNAi或通过钙蛋白酶肽或PD 150606抑制MCF 7/CEACAM 1 -4S细胞中的CAPN 9表达抑制了管腔形成。将CAPN 9转染到野生型MCF 7细胞中恢复管腔形成，证明钙蛋白酶-9可能在管腔形成中起关键作用。此外，我们证明了细胞凋亡相关激酶PKC-δ在3D培养中生长的野生型CEACAM 1 -4S转染的MCF 7细胞中仅在管腔形成期间被蛋白水解裂解激活，并且管腔形成被针对PKC-δ的RNAi或PKC-δ抑制剂rottlerin抑制。
CEACAM1-4S (carcinoembryonic antigen-related cell adhesion molecule 1) is a type I membrane protein with a short (12 amino acid) cytoplasmic tail. Wild type CEACAM1-4S transfected MCF7 cells form glands with lumena when grown in 3D culture, while null mutations of two putative phosphorylation sites (T457A and S459A) in the cytoplasmic domain fail to undergo lumen formation. When gene chip analysis was performed on mRNA isolated from both wild type and T457A,S459A mutated CEACAM1-4S transfected MCF7 cells grown in 3D culture, calpain-9 (CAPN9) was identified out of over 400 genes with a >2 log 2 difference as a potential inducer of lumen formation. Inhibition of CAPN9 expression in MCF7/CEACAM1-4S cells by RNAi or by calpeptin or PD150606 inhibited lumen formation. Transfection of CAPN9 into wild type MCF7 cells restores lumen formation demonstrating that calpain-9 may plays a critical role in lumen formation. Additionally, we demonstrate that the apoptosis related kinase, PKC-δ is activated by proteolytic cleavage during lumen formation exclusively in wild type CEACAM1-4S transfected MCF7 cells grown in 3D culture, and that lumen formation is inhibited by either RNAi to PKC-δ or by the PKC-δ inhibitor rottlerin.
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影响因子：
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作者：
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影响因子：
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