Skp1蛋白在非小细胞肺癌组织和外周血中的表达水平及其临床意义
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摘要
目的:检测S期激酶相关蛋白1(Skp1)在非小细胞肺癌(NSCLC)组织和外周血中的表达水平并探讨其临床意义。方法:采用Western blot检测34例NSCLC组织及相应癌旁组织中Skp1蛋白的表达水平。采用组织芯片和免疫组织化学染色检测Skp1在86例NSCLC组织及相应癌旁组织中的表达情况,并分析Skp1蛋白的表达与NSCLC临床病理指标及患者预后的关系。采用酶联免疫吸附试验(ELISA)检测39例NSCLC患者及27例健康人血浆中的Skp1蛋白水平。结果:Western blot和免疫组化法分析结果显示Skp1蛋白在NSCLC组织中的表达水平均显著高于相应癌旁组织(P<0.001)。ELISA法分析结果显示NSCLC病人血浆中的Skp1蛋白表达水平显著高于健康人群(P=0.003)。Kaplan-Meier单因素生存分析显示Skp1蛋白在NSCLC组织中的高表达与中晚期(Ⅱ+Ⅲ)NSCLC患者的不良预后显著相关(P=0.001),多因素Cox回归分析显示Skp1蛋白能够作为影响中晚期(Ⅱ+Ⅲ)NSCLC患者预后的独立危险因素。Skp1蛋白表达水平与病理类型、临床分期、分化程度、淋巴结转移等因素无显著相关关系(P>0.05)。结论:Skp1蛋白在NSCLC患者中高表达,并且Skp1蛋白在NSCLC组织中的高表达与中晚期(Ⅱ+Ⅲ)NSCLC患者的不良预后显著相关,可作为中晚期NSCLC患者预后的独立危险因素。
OBJECTIVE: To detect the protein expression of Skpl(S-phase kinase-associated protein 1)in non-small cancer lung cancer(NSCLC) tissues and peripheral blood, and to explore its clinical significance.METHODS : Expressions of Skpl protein in 34 NSCLC tissues and 34 adjacent tissues were detected by Western Blot. In addition, the expressions in 86 NSCLC tissues and 86 adjacent tissues were detected by tissue microarray and immunohistochemical staining method. The plasma expression level of Skp1 in 39 NSCLC patients and 27 healthy person was detected by ELISA(enzyme linked immunesorbent assay). The relationships between Skpl expression with clinicopathological parameters and prognosis of NSCLC patients were analyzed.RESULTS : The expression level of Skpl protein in NSCLC tissues was significantly higher than that in adjacent tissues by both Western blot and immunohistochemical staining methods(P<0.01). The plasma levels in NSCLC patients were significantly higher than that in healthy person(P=0.003). The simple factors analyses show that over-expression of Skpl protein in NSCLC tissues was significantly associated with poor prognosis in the middle to late cancer stages(Ⅱ + Ⅲ)(P=0.001),and the multiple factors Cox regression analysis showed that Skpl protein could be used as the independent risk factors for the middle to late stages(Ⅱ +Ⅲ). The expression of Skpl protein had no correlation with pathological type, clinical stage, pathological grade, lymph node metastasis and other factors(P>0.05). CONCLUSION: Skpl protein was over-expressed in NSCLC tissues and plasma from patients, and the over-expression of Skpl protein in NSCLC tissues was significantly associated with poor prognosis in the middle to late cancer stages(Ⅱ+Ⅲ). Therefore, the over-expression can be considered for use as an independent risk factor for the middle to late stages among NSCLC patients.
OBJECTIVE: To detect the protein expression of Skpl(S-phase kinase-associated protein 1)in non-small cancer lung cancer(NSCLC) tissues and peripheral blood, and to explore its clinical significance.METHODS : Expressions of Skpl protein in 34 NSCLC tissues and 34 adjacent tissues were detected by Western Blot. In addition, the expressions in 86 NSCLC tissues and 86 adjacent tissues were detected by tissue microarray and immunohistochemical staining method. The plasma expression level of Skp1 in 39 NSCLC patients and 27 healthy person was detected by ELISA(enzyme linked immunesorbent assay). The relationships between Skpl expression with clinicopathological parameters and prognosis of NSCLC patients were analyzed.RESULTS : The expression level of Skpl protein in NSCLC tissues was significantly higher than that in adjacent tissues by both Western blot and immunohistochemical staining methods(P<0.01). The plasma levels in NSCLC patients were significantly higher than that in healthy person(P=0.003). The simple factors analyses show that over-expression of Skpl protein in NSCLC tissues was significantly associated with poor prognosis in the middle to late cancer stages(Ⅱ + Ⅲ)(P=0.001),and the multiple factors Cox regression analysis showed that Skpl protein could be used as the independent risk factors for the middle to late stages(Ⅱ +Ⅲ). The expression of Skpl protein had no correlation with pathological type, clinical stage, pathological grade, lymph node metastasis and other factors(P>0.05). CONCLUSION: Skpl protein was over-expressed in NSCLC tissues and plasma from patients, and the over-expression of Skpl protein in NSCLC tissues was significantly associated with poor prognosis in the middle to late cancer stages(Ⅱ+Ⅲ). Therefore, the over-expression can be considered for use as an independent risk factor for the middle to late stages among NSCLC patients.
引文
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[17]XU M,YANG X,ZHAO J,et al.High expression of Cullin1 indicates poor prognosis for NSCLC patients[J].Pathol Res Pract,2014,210(7):397-401.
[18]HUANG Y F,ZHANG Z,ZHANG M,et al.CUL1promotes breast cancer metastasis through regulating EZH2-induced the autocrine expression of the cytokines CXCL8 and IL11[J].Cell Death Dis,2018,10(1):2.
[19]KIMBREL E A,KUNG A L.The F-box protein betaTrCp1/Fbw1a interacts with p300 to enhance beta-catenin transcriptional activity[J].J Biol Chem,2009,284(19):13033-13044.
[20]WANG J Y,LIU G Z,WILMOTT J S,et al.Skp2-mediated stabilization of MTH1 promotes survival of melanoma cells upon oxidative stress[J].Cancer Res,2017,77(22):6226-6239.
[21]TAKANAMI I.The prognostic value of overexpression of Skp2 mRNA in non-small cell lung cancer[J].Oncol Rep,2005,13(4):727-731.
[22]ZHANG Y,ZVI Y S,BATKO B,et al.Down-regulation of Skp2 expression inhibits invasion and lung metastasis in osteosarcoma[J].Sci Rep 2018,8(1):14294.
[23]DING L,LI R,SUN R,et al.S-phase kinaseassociated protein 2 promotes cell growth and motility in osteosarcoma cells[J].Cell Cycle,2017,16(16):1547-1555.
[24]DING L,LI R,HAN X,et al.Inhibition of Skp2suppresses the proliferation and invasion of osteosarcoma cells[J].Oncol Rep,2017,38(2):933-940.
[25]LIU W,WANG Y,ZHANG C,et al.Cullin1 is upregulated and associated with poor patients'survival in hepatocellular carcinoma[J].Int J Clin Exp Pathol,2015,8(4):4001-4007.
[26]MIGITA K,TAKAYAMA T,MATSUMOTO S,et al.Prognostic impact of RING box protein-1(RBX1)expression in gastric cancer[J].Gastric Cancer,2014,17(4):601-609.
[27]RUAN D,HE J,LI C F,et al.Skp2 deficiency restricts the progression and stem cell features of castration-resistant prostate cancer by destabilizing Twist[J].Oncogene,2017,36(30):4299-4310.
[2]TARTARI R F,ULBRICH-KULCZYNSKI J M,FILHOA F.Measurement of mid-arm muscle circumference and prognosis in stage IV non-small cell lung cancer patients[J].Oncol Lett,2013,5(3):1063-1067.
[3]HUSSAIN M,LU Y,LIU Y Q,et al.Skp1:Implications in cancer and SCF-oriented anti-cancer drug discovery[J].Pharmacol Res,2016,111:34-42.
[4]KUHN D J,CHEN Q,VOORHEES P M,et al.Potent activity of carfilzomib,a novel,irreversible inhibitor of the ubiquitin-proteasome pathway,against preclinical models of multiple myeloma[J].Blood,2007,110(9):3281-3290.
[5]RICHARDSON P G,SONNEVELD P,SCHUSTER MW,et al.Bortezomib or high-dose dexamethasone for relapsed multiple myeloma[J].N Engl J Med,2005,352(24):2487-2498.
[6]HERSHKO A,CIECHANOVER A.The ubiquitin system[J].Annu Rev Biochem,1998,67:425-479.
[7]YOSHIDA Y,MURAKAMI A,TANAKA K.Skp1stabilizes the conformation of F-box proteins[J].Biochem Biophys Res Commun,2011,410(1):24-28.
[8]SKAAR J R,PAGAN J K,PAGANO M.Mechanisms and function of substrate recruitment by F-box proteins[J].Nat Rev Mol Cell Biol,2013,14(6):369-381.
[9]GSTAIGER M,MARTI A,KREK W.Association of human SCF(SKP2)subunit p19(SKP1)with interphase centrosomes and mitotic spindle poles[J].Exp Cell Res,1999,247(2):554-562.
[10]CHANDRA DANTU S,NATHUBHAI KACHARIYA N,KUMAR A.Molecular dynamics simulations elucidate the mode of protein recognition by Skp1 and the F-box domain in the SCF complex[J].Proteins,2016,84(1):159-171.
[11]BAI C,SEN P,HOFMANN K,et al.Skp1 connects cell cycle regulators to the ubiquitin proteolysis machinery through a novel motif,the F-box[J].Cell,1996,86(2):263-274.
[12]LIU Y Q,WANG X L,CHENG X,et al.Skp1 in lung cancer:clinical significance and therapeutic efficacy of its small molecule inhibitors[J].Oncotarget,2015,6(33):34953-34967.
[13]WANG Z,LIU P,INUZUKA H,et al.Roles of F-box proteins in cancer[J].Nat Rev Cancer,2014,14(4):233-247.
[14]REMMELE W,STEGNER H E.Recommendation for uniform definition of an immunoreactive score(IRS)for immunohistochemical estrogen receptor detection(ER-ICA)in breast cancer tissue[J].Pathologe,1987,8(3):138-140.
[15]WANG X,ZHANG S,ZHANG T,et al.Prognostic values of F-box members in breast cancer:an online database analysis and literature review[J].Biosci Rep,2018.pii:BSR20180949.
[16]YU Z K,GERVAIS J L,ZHANG H.Human CUL-1associates with the SKP1/SKP2 complex and regulates p21(CIP1/WAF1)and cyclin D proteins[J].Proc Natl Acad Sci USA,1998,95(19):11324-11329.
[17]XU M,YANG X,ZHAO J,et al.High expression of Cullin1 indicates poor prognosis for NSCLC patients[J].Pathol Res Pract,2014,210(7):397-401.
[18]HUANG Y F,ZHANG Z,ZHANG M,et al.CUL1promotes breast cancer metastasis through regulating EZH2-induced the autocrine expression of the cytokines CXCL8 and IL11[J].Cell Death Dis,2018,10(1):2.
[19]KIMBREL E A,KUNG A L.The F-box protein betaTrCp1/Fbw1a interacts with p300 to enhance beta-catenin transcriptional activity[J].J Biol Chem,2009,284(19):13033-13044.
[20]WANG J Y,LIU G Z,WILMOTT J S,et al.Skp2-mediated stabilization of MTH1 promotes survival of melanoma cells upon oxidative stress[J].Cancer Res,2017,77(22):6226-6239.
[21]TAKANAMI I.The prognostic value of overexpression of Skp2 mRNA in non-small cell lung cancer[J].Oncol Rep,2005,13(4):727-731.
[22]ZHANG Y,ZVI Y S,BATKO B,et al.Down-regulation of Skp2 expression inhibits invasion and lung metastasis in osteosarcoma[J].Sci Rep 2018,8(1):14294.
[23]DING L,LI R,SUN R,et al.S-phase kinaseassociated protein 2 promotes cell growth and motility in osteosarcoma cells[J].Cell Cycle,2017,16(16):1547-1555.
[24]DING L,LI R,HAN X,et al.Inhibition of Skp2suppresses the proliferation and invasion of osteosarcoma cells[J].Oncol Rep,2017,38(2):933-940.
[25]LIU W,WANG Y,ZHANG C,et al.Cullin1 is upregulated and associated with poor patients'survival in hepatocellular carcinoma[J].Int J Clin Exp Pathol,2015,8(4):4001-4007.
[26]MIGITA K,TAKAYAMA T,MATSUMOTO S,et al.Prognostic impact of RING box protein-1(RBX1)expression in gastric cancer[J].Gastric Cancer,2014,17(4):601-609.
[27]RUAN D,HE J,LI C F,et al.Skp2 deficiency restricts the progression and stem cell features of castration-resistant prostate cancer by destabilizing Twist[J].Oncogene,2017,36(30):4299-4310.