Cis-hydroxyproline-induced inhibition of pancreatic cancer cell growth is mediated by endoplasmic reticulum stress

瞄准：在老鼠上调查 cis-hydroxyproline (CHP ) 的生物效果，并且检验内在的分子的机制胰腺的癌房间线 DSL6A。方法：DSL6A 房间增长上的 CHP 的效果被使用 BrdU 加入估计。焦点的粘附激酶(FAK ) 的表示被西方的弄污和免疫荧光描绘。endoplasmic 感应(嗯) 应力被使用 RT-PCR 并且为葡萄糖相关的 protein-78 (GRP78 ) 和生长拘捕和 DNA 的西方的弄污调查可诱导的基因(GADD153 ) 。房间生存能力通过基于 DSL6A 房间的减小潜力测量新陈代谢的活动被决定。Apoptosis 被 caspase-3 激活的察觉和多形核白细胞(自动数据处理核糖) 的劈开分析象 DNA laddering 一样的聚合酶(PARP ) 。结果：除了增长的抑制，有 CHP 的孵化从焦点的粘附导致了 FAK 的解朊的劈开和酶的 delocalisation，由房间坚持的损失列在后面。同时，我们能显示出 GRP78 和 GADD153 的增加的表情，显示在 DSL6A 房间线的 ER 压力串联的调停 CHP 的激活。有 CHP 的 DSL6A 房间的延长孵化最后导致了 apoptotic 细胞死亡。在 L 脯氨酸旁边，由一个广谱朊酶禁止者的增加的细胞内部的解朊作用的抑制能在细胞的功能和分子的过程上废除 CHP 的效果。相反，阻碍执行 apoptosis caspases 的活动没在调停 CHP 的房间损坏上有影响。结论：我们的数据建议开始嗯由 CHP 的压力机械导致细胞内部的解朊的进程的激活包括 caspase 独立的 FAK 降级，导致损坏胰腺的癌房间。

AIM： To investigate the biological effects of cishydroxyproline （CHP） on the rat pancreatic carcinoma cell line DSL6A, and to examine the underlying molecular mechanisms. METHODS： The effect of CHP on DSL6A cell proliferation was assessed by using BrdU incorporation. The expression of focal adhesion kinase （FAK） was characterized by Western blotting and immunofluorescence. Induction of endoplasmic reticulum （ER） stress was investigated by using RT-PCR and Western blotting for the glucose-related protein-78 （GRP78） and growth arrest and DNA inducible gene （GADD153）. Cell viability was determined through measuring the metabolic activity based on the reduction potential of DSL6A cells. Apoptosis was analyzed by detection of caspase-3 activation and cleavage of poly（ADP-ribose） polymerase （PARP） as well as DNA laddering. RESULTS： In addition to inhibition of proliferation, incubation with CHP induced proteolytic cleavage of FAK and a delocalisation of the enzyme from focal adhesions, followed by a loss of cell adherence. Simultaneously, we could show an increased expression of GRP78 and GADD153, indicating a CHP-mediated activation of the ER stress cascade in the DSL6A cell line. Prolonged incubation of DSL6A cells with CHP finally resulted in apoptotic cell death. Beside L-proline, the inhibition of intracellular proteolysis by addition of a broad spectrum protease inhibitor could abolish the effects of CHP on cellular functions and the molecular processes. In contrast, impeding the activity of apoptosis-executing caspases had no influence on CHP-mediated cell damage. CONCLUSION： Our data suggest that the initiation of ER stress machinery by CliP leads to an activation of intracellular proteolytic processes, including caspaseindependent FAK degradation, resulting in damaging pancreatic carcinoma cells.