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首页> 外文期刊>Pancreas >Investigation of the mechanisms by which EB1089 abrogates apoptosis induced by 9-cis retinoic acid in pancreatic cancer cells.
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Investigation of the mechanisms by which EB1089 abrogates apoptosis induced by 9-cis retinoic acid in pancreatic cancer cells.

机译:研究EB1089消除9-顺式视黄酸诱导的胰腺癌细胞凋亡的机制。

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OBJECTIVES: Previous research has shown that the retinoid 9-cis retinoic acid (RA) promotes apoptosis in pancreatic cancer cells. The vitamin D analog EB1089 does not. Furthermore, cotreatment of cells with 9-cis RA and EB1089 abrogates apoptosis. To explain this, we studied the regulation of proteins involved in apoptotic signaling pathways in pancreatic cancer cells. METHODS: The pancreatic adenocarcinoma cell line T3M4 was used. Cell proliferation was measured using the SRB protein dye assay. Induction of apoptosis was evaluated using an ELISA assay. Caspase activation was detected using a colorimetric assay based on cleavage of a caspase-associated substrate. Regulation of protein levels and posttranslational events were detected using immunoblotting. RESULTS: We confirm that EB1089 diminishes apoptosis induced by 9-cis RA in T3M4 cells. We extend the study to show that EB1089 abrogates increases, induced by 9-cis RA, in caspase activation, p27Kip1 protein levels, Bim and Bax protein levels and in Bax/Bcl2 ratio. In addition, the CDKI p21Waf1 and CAII, a differentiation marker for pancreatic cancer cells are also differentially regulated. CONCLUSIONS: These results suggest that the inhibitory effects of EB1089 on 9-cis RA-induced apoptosis lie upstream of caspase activation and could be associated with reduction of p27Kip1 protein levels.
机译:目的:以前的研究表明,类维生素A 9-顺式视黄酸(RA)促进胰腺癌细胞的凋亡。维生素D类似物EB1089没有。此外,将细胞与9-顺式RA和EB1089共同处理可消除凋亡。为了解释这一点,我们研究了胰腺癌细胞凋亡信号通路中涉及的蛋白质调控。方法:使用胰腺腺癌细胞系T3M4。使用SRB蛋白染料测定法测量细胞增殖。使用ELISA测定法评估凋亡的诱导。使用基于比色半胱天冬酶相关底物的裂解的比色测定法检测半胱天冬酶的活化。使用免疫印迹检测蛋白质水平和翻译后事件的调节。结果:我们证实EB1089减少了9-顺式RA诱导的T3M4细胞凋亡。我们扩展了研究范围,以显示由9-顺式RA诱导的EB1089废除在caspase激活,p27Kip1蛋白水平,Bim和Bax蛋白水平以及Bax / Bcl2比方面增加。此外,CDKI p21Waf1和CAII(胰腺癌细胞的分化标记)也受到差异调节。结论:这些结果表明,EB1089对9-顺式RA诱导的细胞凋亡的抑制作用位于caspase激活的上游,并且可能与p27Kip1蛋白水平的降低有关。

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