钙周期素结合蛋白促进胃癌细胞迁移侵袭和MMP-2、MMP-9、p-ERK1/2、p-AKT水平上调
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摘要
目的探讨钙周期素结合蛋白(calcyclin binding protein/Siah-1-interacting protein, CacyBP/SIP)对胃癌细胞侵袭迁移的影响和潜在机制。方法采用免疫组织化学和Western blot方法检测不同T分期胃癌组织中CacyBP/SIP水平;Western blot检测胃癌细胞中CacyBP/SIP水平;MKN-45细胞转染si-CacyBP/SIP与Ad-CacyBP/SIP后,细胞划痕实验检测细胞迁移情况,Transwell细胞侵袭实验检测细胞侵袭情况,Western blot检测MMP-2、MMP-9和p-ERK1/2、p-AKT水平。结果CacyBP/SIP在胃癌组织和胃癌细胞中高表达;胃癌组织中CacyBP/SIP表达水平与T分期呈正相关;敲减CacyBP/SIP抑制MKN-45细胞的迁移侵袭能力和MMP-2、MMP-9、p-ERK1/2、p-AKT蛋白表达水平;过表达CacyBP/SIP促进MKN-45细胞迁移侵袭能力和MMP-2、MMP-9、p-ERK1/2、p-AKT蛋白表达水平。结论 CacyBP/SIP对胃癌转移侵袭能力的促进作用可能与其上调MMP-2、MMP-9、p-ERK1/2、p-AKT水平有关。
Objective To investigate the effect of calcyclin binding protein/Siah-1-interacting protein(CacyBP/SIP) on invasion and migration in gastric cancer cells and its potential mechanism. Methods The expression levels of CacyBP/SIP in different T stages of gastric cancer tissues were detected by Western blot and immunohistochemical analysis; The expression of CacyBP/SIP in gastric cancer cells were detected by Western blot. After transfected with si-CacyBP/SIP or Ad-CacyBP/SIP in MKN-45 cells, cell migration was measured by in vitro cell scratch assay and cell invasion was measured by transwell assay, the expression levels of MMP-2, MMP-9, p-ERK1/2 and p-AKT were detected by Western blot. Results Cacy BP/SIP was highly expressed in gastric cancer tissues and gastric cancer cell lines. Especially, CacyBP/SIP expression level was positively correlated with T stage in gastric cancer tissues. Knockdown of CacyBP/SIP inhibited the abilities of invasion and migration, as well as the expression levels of MMP-2, MMP-9, p-ERK1/2 and p-AKT in MKN-45 cells; while overexpression of CacyBP/SIP promoted the abilities of invasion and migration as well as the expressions of MMP-2, MMP-9, p-ERK1/2 and p-AKT in MKN-45 cells. Conclusion The promotive effect of CacyBP/SIP on the abilities of invasion and migration may be related to the up-regulation of MMP-2, MMP-9, p-ERK1/2 and p-AKT levels in gastric cancer cells.
Objective To investigate the effect of calcyclin binding protein/Siah-1-interacting protein(CacyBP/SIP) on invasion and migration in gastric cancer cells and its potential mechanism. Methods The expression levels of CacyBP/SIP in different T stages of gastric cancer tissues were detected by Western blot and immunohistochemical analysis; The expression of CacyBP/SIP in gastric cancer cells were detected by Western blot. After transfected with si-CacyBP/SIP or Ad-CacyBP/SIP in MKN-45 cells, cell migration was measured by in vitro cell scratch assay and cell invasion was measured by transwell assay, the expression levels of MMP-2, MMP-9, p-ERK1/2 and p-AKT were detected by Western blot. Results Cacy BP/SIP was highly expressed in gastric cancer tissues and gastric cancer cell lines. Especially, CacyBP/SIP expression level was positively correlated with T stage in gastric cancer tissues. Knockdown of CacyBP/SIP inhibited the abilities of invasion and migration, as well as the expression levels of MMP-2, MMP-9, p-ERK1/2 and p-AKT in MKN-45 cells; while overexpression of CacyBP/SIP promoted the abilities of invasion and migration as well as the expressions of MMP-2, MMP-9, p-ERK1/2 and p-AKT in MKN-45 cells. Conclusion The promotive effect of CacyBP/SIP on the abilities of invasion and migration may be related to the up-regulation of MMP-2, MMP-9, p-ERK1/2 and p-AKT levels in gastric cancer cells.
引文
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[16] Filipek A. S100A6 and CacyBP/SIP-two proteins discoveredinehrlichascitestumorcellsthatarepotentiallyinvolved in the degradation ofβ-catenin. Chemotherapy, 2006,52(1):32-34.
[17] AmanoS,AkutsuN,Matsunaga Y,etal.Importanceof balance between extracellular matrix synthesis and degradation in basement membrane formation. Exp Cell Res, 2001,271(2):249-262.
[18] LiaoCL,Chu YL,LinHY,etal.Bisdemethoxycurcumin SuppressesMigrationandInvasionofHumanCervical Cancer HeLa CellsviaInhibition of NF-?B,MMP-2and-9 Pathways. Anticancer Res, 2018, 38(7):3989-3997.
[19] Yan S, Li A, Liu Y. CacyBP/SIPinhibits the migration and invasion behaviors of glioblastoma cells through activating Siah1 mediated ubiquitination and degradation of cytoplasmic p27. Cell Biol Int, 2018, 42(2):216-226.
[20] Schneider G, Nieznanski K, Jozwiak J, et al. Tubulin binding protein, CacyBP/SIP, induces actin polymerization and may link actin and tubulin cytoskeletons. Biochim Biophys Acta, 2010, 1803(11):1308-1317.
[21] Peng Y, Lin J, Ma J, et al. Upregulation of S100A6 in patients with endometriosis and its role in ectopic endometrial stromal cells. Gynecol Endocrinol, 2018, 34(9):815-820.
[22] Wang N1, Ma Q, Wang Y, et al. CacyBP/SIP expression is involved in the clinical progression of breast cancer. World J Surg, 2010, 34(11):2545-2452.
[23] KilanczykE,FilipekS,JastrzebskaB,etal.CacyBP/SIPbindsERK1/2andaffectstranscriptionalactivityof Elk-1.BiochemBiophysResCommun,2009,380(1):54-59.
[24] Guo S, Yu L, Cheng Y, et al. PDGFRβtriggered by bFGF promotestheproliferationandmigrationofendothelial progenitor cells via p-ERK signalling. Cell Biol Int, 2012,36(10):945-950.
[25] Henderson V,SmithB,BurtonLJ,etal.Snailpromotes cell migration through PI3K/AKT-dependent Rac1 activationaswellasPI3K/AKT-independentpathwaysduring prostate cancer progression. Cell Adh Migr, 2015, 9(4):255-264.
[2]Deng Q, He B, Liu X, et al. Prognostic value of pre-operative inflammatory response biomarkers in gastric cancer patients and the construction of a predictive model. J Transl Med,2015, 13(1):409.
[3]Kato T, Suzuki K, Muto Y, et al. Multiple primary malignancies involving primary sporadic colorectal cancer in Japan:incidence of gastric cancer with colorectal cancer patients maybehigherthanpreviouslyrecognized. WorldJSurg Oncol, 2015, 13(1):432.
[4]Chen F, Zhuang M, Zhong C, et al. Baicalein reverses hypoxia-induced 5-FU resistance in gastric cancer AGS cells through suppression of glycolysis and the PTEN/Akt/HIF-1αsignaling pathway. Oncol Rep, 2015, 33(1):457-463.
[5]Zhang X, Park JS, Park KH, et al. PTEN Deficiency as a Predictive Biomarker of Resistance to HER2-Targeted Therapy in Advanced Gastric Cancer. Oncology, 2015, 88(2):76-85.
[6]ZhaiH,Shi Y,ChenX,etal.CacyBP/SIPpromotesthe proliferation of coloncancercells. PLoS One, 2017, 12(2):e0169959.
[7]Fu C, Wan Y, Shi H, et al. Expression and regulation ofCacy BP/SIPin chronic lymphocytic leukemia cell balances of cell proliferation with apoptosis. J Cancer Res Clin Oncol,2016, 142(4):741-748.
[8]Shi H, Gao Y, Tang Y, et al. CacyBP/SIPprotein is important for the proliferation of human glioma cells. IUBMB Life,2014, 66(4):286-291.
[9]Tang Y, Zhan W, Cao T, et al. CacyBP/SIPinhibits Doxourbicin-induced apoptosis of glioma cells due to activation of ERK1/2. IUBMB Life, 2016, 68(3):211-219.
[10] Chen X, Mo P, Li X, et al. CacyBP/SIPprotein promotes proliferation and G1/S transition of human pancreatic cancer cells. Mol Carcinog, 2011, 50(10):804-810.
[11] Chen Y, Zhang K, Wang X, et al. Cell cycle-dependent translocation and regulatory mechanism ofCacy BP/SIPingastric cancer cells. Anticancer Drugs, 2018, 29(1):19-28.
[12] Zhai H, Meng J, Jin H, et al. Role of theCacyBP/SIPprotein ingastric cancer. Oncol Lett, 2015, 9(5):2031-2035.
[13] Li W,LiS,DengL,etal.DecreasedMT1-MMPingastric cancer suppressed cell migration and invasion via regulating MMPs and EMT. Tumour Biol, 2015, 36(9):6883-6889.
[14] Tapia O, Riquelme I, Leal P, et al. The PI3K/AKT/mTOR pathway is activated in gastric cancer with potential prognosticandpredictivesignificance. Virchows Arch,2014,465(1):25-33.
[15] Cai H, Chen X, Zhang J, et al. 18β-glycyrrhetinic acid inhibits migration and invasion of human gastric cancer cells via the ROS/PKC-α/ERK pathway. J Nat Med, 2018, 72(1):252-259.
[16] Filipek A. S100A6 and CacyBP/SIP-two proteins discoveredinehrlichascitestumorcellsthatarepotentiallyinvolved in the degradation ofβ-catenin. Chemotherapy, 2006,52(1):32-34.
[17] AmanoS,AkutsuN,Matsunaga Y,etal.Importanceof balance between extracellular matrix synthesis and degradation in basement membrane formation. Exp Cell Res, 2001,271(2):249-262.
[18] LiaoCL,Chu YL,LinHY,etal.Bisdemethoxycurcumin SuppressesMigrationandInvasionofHumanCervical Cancer HeLa CellsviaInhibition of NF-?B,MMP-2and-9 Pathways. Anticancer Res, 2018, 38(7):3989-3997.
[19] Yan S, Li A, Liu Y. CacyBP/SIPinhibits the migration and invasion behaviors of glioblastoma cells through activating Siah1 mediated ubiquitination and degradation of cytoplasmic p27. Cell Biol Int, 2018, 42(2):216-226.
[20] Schneider G, Nieznanski K, Jozwiak J, et al. Tubulin binding protein, CacyBP/SIP, induces actin polymerization and may link actin and tubulin cytoskeletons. Biochim Biophys Acta, 2010, 1803(11):1308-1317.
[21] Peng Y, Lin J, Ma J, et al. Upregulation of S100A6 in patients with endometriosis and its role in ectopic endometrial stromal cells. Gynecol Endocrinol, 2018, 34(9):815-820.
[22] Wang N1, Ma Q, Wang Y, et al. CacyBP/SIP expression is involved in the clinical progression of breast cancer. World J Surg, 2010, 34(11):2545-2452.
[23] KilanczykE,FilipekS,JastrzebskaB,etal.CacyBP/SIPbindsERK1/2andaffectstranscriptionalactivityof Elk-1.BiochemBiophysResCommun,2009,380(1):54-59.
[24] Guo S, Yu L, Cheng Y, et al. PDGFRβtriggered by bFGF promotestheproliferationandmigrationofendothelial progenitor cells via p-ERK signalling. Cell Biol Int, 2012,36(10):945-950.
[25] Henderson V,SmithB,BurtonLJ,etal.Snailpromotes cell migration through PI3K/AKT-dependent Rac1 activationaswellasPI3K/AKT-independentpathwaysduring prostate cancer progression. Cell Adh Migr, 2015, 9(4):255-264.