SCP1对人乳腺癌细胞迁移和增殖的影响及其机制
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摘要
SCP1是一种膜定位的磷酸酶,可以去磷酸化RNA聚合酶Ⅱ并沉默神经元基因.目的:探讨SCP1对人乳腺癌MDA-MB-231细胞迁移和增殖能力的影响,并研究其机制.方法:荧光定量PCR法检测多种癌细胞中SCP1的表达;体外划痕实验检测SCP1对MDA-MB-231细胞迁移能力的影响;Transwell TM细胞侵袭实验检测SCP1对MDA-MB-231细胞侵袭能力的影响;MTT法检测SCP1对MDA-MB-231细胞增殖能力的影响;Western blot法检测SCP1对细胞信号通路分子p-AKT表达的影响.结果:SCP1多种癌细胞中均有表达;SCP1能抑制MDA-MB-231细胞的迁移及增殖能力;抑制SCP1的表达能显著上调MDA-MB-231细胞p-AKT的表达.结论:SCP1可能通过去磷酸化AKT抑制MDA-MB-231细胞迁移和侵袭.
SCP1 as a membrane located phosphatases acts globally to silence neuronal genes and to affect the dephosphorylation of RNA PolⅡ.Objective:To investigate the effect and the relevant molecular mechanisms of SCP1 on the migration and proliferation of human breast cancer MDA-MB-231 cell.Methods:The expression of SCP1 in various cancer cells was measured by qRT-PCR.MTT assay was used to evaluate the proliferation capacity in different MDA-MB-231 breast cancer cells.The effects of SCP1 on the expression of p-AKT in MDA-MB-231 cells were examined by Western blotting.In vitro wound healing assay and Transwell TMassay were utilized to measure the effects of SCP1 on the migration and invasion capability of MDA-MB-231 cells.Results:Suppressed SCP1 significantly promoted the migration and proliferation of MDA-MB-231 cells.Inhibition of SCP1 did not affect the expression of total AKT,whereas the phosphorylated AKT level was markedly up-regulated.Conclusion:SCP1 could inhibit the migration and proliferation of human breast cancer MDA-MB-231 cell via dephosphorylation of AKT.
SCP1 as a membrane located phosphatases acts globally to silence neuronal genes and to affect the dephosphorylation of RNA PolⅡ.Objective:To investigate the effect and the relevant molecular mechanisms of SCP1 on the migration and proliferation of human breast cancer MDA-MB-231 cell.Methods:The expression of SCP1 in various cancer cells was measured by qRT-PCR.MTT assay was used to evaluate the proliferation capacity in different MDA-MB-231 breast cancer cells.The effects of SCP1 on the expression of p-AKT in MDA-MB-231 cells were examined by Western blotting.In vitro wound healing assay and Transwell TMassay were utilized to measure the effects of SCP1 on the migration and invasion capability of MDA-MB-231 cells.Results:Suppressed SCP1 significantly promoted the migration and proliferation of MDA-MB-231 cells.Inhibition of SCP1 did not affect the expression of total AKT,whereas the phosphorylated AKT level was markedly up-regulated.Conclusion:SCP1 could inhibit the migration and proliferation of human breast cancer MDA-MB-231 cell via dephosphorylation of AKT.
引文
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[2]Yeo M.A novel RNA polymeraseⅡC-terminal domain phosphatase that preferentially dephosphorylates serine[J].J Boil Chem,2003,278(28):26078-26085.
[3]Kamenski T,Heilmeier S,Meinhart A,et al.Structure and mechanism of RNA polymerase II CTD phosphatases[J].Molecular Cell,2004,15(3):399-407.
[4]Yeo M,Lee SK,Lee B,et al.Small CTD phosphatases function in silencing neuronal gene expression[J].Science,2005,307(5709):596-600.
[5]Sapkota G,Knockaert M,Alarcon C,et al.Dephosphorylation of the linker regions of Smad1and Smad2/3by small C-terminal domain phosphatases has distinct outcomes for bone morphogenetic protein and transforming growth factor-beta pathways[J].J Boil Chem,2006,281(52):40412-40419.
[6]Thompson J,Lepikhova T,Teixido-Travesa N,et al.Small carboxyl-terminal domain phosphatase 2attenuates androgen-dependent transcription[J].Embo,2006,25(12):2757-2767.
[7]廖鹏.磷酸酯酶SCP1调控c-Myc蛋白去磷酸化及相关生物学功能的研究和SCP1细胞膜定位的发现及其棕榈酰化修饰的研究[D].上海:华东师范大学,2012.
[8]郑莹,吴春晓,张敏璐.乳腺癌在中国的流行状况和疾病特征[J].中国癌症杂志,2013,23(8):561-569.
[9]赵文静,旺建伟,隋方宇,等.乳腺增生病与乳腺癌病因相关性研究[J].中医药学报,2015,43(3):28-30.
[10]李勇杰,于庆龙,潘际刚,等.酒精对乳腺癌细胞表皮生长因子受体-钙激活中性蛋白酶通路及细胞迁移的影响[J].中国癌症杂志,2016,26(10):820-825.
[11]Hemmings B A,Restuccia D F.The PI3K-PKB/Akt pathway[J].Cold Spring Harbor Perspectives in Biology,2015,7(4):95-101.
[12]Liao P,Wang W,Li Y,et al.Palmitoylated SCP1is targeted to the plasma membrane and negatively regulates angiogenesis[J].Elife,2017,6:38-42.
[13]Testa J R,Bellacosa A.AKT plays a central role in tumorigenesis[J].Proc Natl Acad Sci USA,2001,98(20):10983-10985.
[14]Okumura N,Yoshida H,Kitagishi Y,et al.PI3K/AKT/PTEN signaling as a molecular target in leukemia angiogenesis[J].Adv Hematol,2012,2012:843085.
[15]刘思旸,邹志鹏,郦俊,等.蛋白激酶蛋白家族的功能研究进展[J].医学研究生学报,2011,24(2):208-211.
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