Over-expression of ubiquitin carboxy terminal hydrolase-L1 induces apoptosis in breast cancer cells.
Over-expression of ubiquitin carboxy terminal hydrolase-L1 induces apoptosis in breast cancer cells.
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DOI：
10.3892/ijo_00000092
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发表时间：
2008-11
影响因子：
5.2
通讯作者：
Liu, Xiu-Ping
中科院分区：
文献类型：
--
作者：
Wang, Wen-Juan;Li, Qing-Quan;Xu, Jing-Da;Cao, Xi-Xi;Li, Hai-Xia;Tang, Feng;Chen, Qi;Yang, Jin-Ming;Xu, Zu-De;Liu, Xiu-Ping
泛素羧基末端水解酶-L1（UCH-L1）属于UCH蛋白酶家族，在泛素-蛋白酶体系统中使泛素-蛋白缀合物去泛素化。前期研究表明，UCH-L1在小鼠视网膜细胞和睾丸生殖细胞中表达，其功能与细胞凋亡有关。但UCH-L1是否与肿瘤细胞凋亡有关尚不清楚。为了阐明UCH-L1在肿瘤细胞中的作用，本研究选择了UCH-L1表达相对高的多药耐药（MDR）人乳腺癌细胞系MCF 7/Adr及其亲本细胞系MCF 7，其表达相对低。将pcDNA3.1-UCH-L1质粒和UCH-L1 siRNA分别转染MCF 7和MCF 7/Adr细胞。通过MTT法、Western blot、Hoechst 33258染色法和流式细胞仪检测，发现UCH-L1过表达可诱导MCF 7细胞凋亡。另一方面，UCH-L1在MCF 7/Adr细胞中的沉默导致相反的效果。此外，为了探讨这些观察结果的机制，我们进一步研究了磷酸化Akt及其下游信号磷酸化I κB-α和其他信号分子包括Fas、Fas-L、Trail、DR 4、DR 5、Bax、细胞色素C、活性caspase-3、磷酸化p53、磷酸化Mdm-2、Bcl-2、Bcl-xL、p21和p27的表达。结果提示，UCH-L1诱导的细胞凋亡过程可能部分通过PI 3 K/ Akt信号通路。我们的研究结果表明，调节泛素化和去泛素化途径可能是一种新的肿瘤治疗方法。
Ubiquitin carboxy terminal hydrolase-L1 (UCH-L1) belongs to the UCH proteases family that deubiquitinates ubiquitin-protein conjugates in the ubiquitin-proteasome system. Previous research showed that UCH-L1 was expressed in mouse retinal cells and testicular germ cells, and its function was associated with apoptosis. But it is still unclear whether UCH-L1 is concerned with apoptosis in tumor cells. In order to clarify the role of UCH-L1 in tumor cells, multi-drug resistance (MDR) human breast carcinoma cell line MCF7/Adr, that expresses relatively high UCH-L1, and its parental cell line MCF7, that expresses relatively low UCH-L1, were chosen for this study. We transfected pcDNA3.1-UCH-L1 plasmid and UCH-L1 siRNA into MCF7 and MCF7/Adr cells, respectively. Using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, Western blot, Hoechst 33258 staining assay and flow cytometry, we found that over-expression of UCH-L1 in MCF7 cells induced apoptosis. On the other hand, silencing of UCH-L1 in MCF7/Adr cells led to the opposite effect. Moreover, to explore the mechanism underling these observations, we further investigated the expression of phospho-Akt and its downstream signal phospho-IκB-α and other signal molecules including Fas, Fas-L, Trail, DR4, DR5, Bax, cytochrome C, active caspase-3, phospho-p53, phospho-Mdm-2, Bcl-2, Bcl-xL, p21 and p27. The results indicated that the process of apoptosis triggered by UCH-L1 is, at least in part, probably through Phosphoinositide 3-kinase (PI3K)/ Akt signal pathway. Our findings suggest that modulating the ubiquitination and deubiquitination pathway could be a novel method for tumor therapy.
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影响因子：
64.5
作者：
Brunet, A;Bonni, A;Greenberg, ME
通讯作者：
Greenberg, ME
影响因子：
10.5
作者：
POLYAK, K;KATO, JY;KOFF, A
通讯作者：
KOFF, A
影响因子：
64.5
作者：
KATO, JY;MATSUOKA, M;SHERR, CJ
通讯作者：
SHERR, CJ
影响因子：
7.5
作者：
Downward, J
通讯作者：
Downward, J
影响因子：
5.7
作者：
Li, Qing-Quan;Wang, Wen-Juan;Xu, Zu-De
通讯作者：
Xu, Zu-De