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神经母细胞瘤 NB2a 和结肠癌细胞 HCT116 中的 CacyBP/SIP 磷酸酶活性。

CacyBP/SIP phosphatase activity in neuroblastoma NB2a and colon cancer HCT116 cells.

机构信息

Nencki Institute of Experimental Biology, 3 Pasteur Street, 02-093 Warsaw, Poland.

出版信息

Biochem Cell Biol. 2012 Aug;90(4):558-64. doi: 10.1139/o2012-011. Epub 2012 Apr 5.

Abstract

Recently, we have reported that CacyBP/SIP could be a novel phosphatase for ERK1/2 kinase. In this work, we analyzed the CacyBP/SIP phosphatase activity toward ERK1/2 in 2 cell lines of different origin. We showed that overexpression of CacyBP/SIP in NB2a cells resulted in a lower level of phosphorylated ERK1/2 (P-ERK1/2) in the nuclear fraction while such overexpression in HCT116 cells had no effect on the level of P-ERK1/2. Moreover, we found that overexpression of CacyBP/SIP resulted in higher phosphatase activity in the nuclear fraction obtained from NB2a cells when compared with HCT116 cells. Using 2-D electrophoresis we showed that the pattern of spots representing CacyBP/SIP differed in these 2 cell lines and was probably due to a different phosphorylation state of this protein. We also established that after overexpression of CacyBP/SIP in NB2a cells, the amount of nuclear β-catenin was low, while it remained high in HCT116 cells. Since NB2a cells have differentiation potential and HCT116 cells do not, our data suggest that different activity of CacyBP/SIP in these 2 cell lines might affect the ERK1/2 pathway in the differentiation or proliferation processes.

摘要

最近,我们报道了 CacyBP/SIP 可能是一种新型的 ERK1/2 激酶磷酸酶。在这项工作中,我们分析了 CacyBP/SIP 对不同来源的 2 种细胞系中 ERK1/2 的磷酸酶活性。我们表明,CacyBP/SIP 在 NB2a 细胞中的过表达导致核部分中磷酸化 ERK1/2(P-ERK1/2)水平降低,而在 HCT116 细胞中过表达则对 P-ERK1/2 水平没有影响。此外,我们发现 CacyBP/SIP 的过表达导致来自 NB2a 细胞的核部分中的磷酸酶活性高于 HCT116 细胞。通过 2-DE 电泳,我们表明在这 2 种细胞系中,代表 CacyBP/SIP 的斑点模式不同,可能是由于该蛋白的不同磷酸化状态所致。我们还确定,在 NB2a 细胞中过表达 CacyBP/SIP 后,核 β-连环蛋白的量较低,而在 HCT116 细胞中则保持较高水平。由于 NB2a 细胞具有分化潜力,而 HCT116 细胞则没有,我们的数据表明,这两种细胞系中 CacyBP/SIP 的不同活性可能会影响 ERK1/2 通路在分化或增殖过程中的作用。

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