摘要
目的:检测亨廷顿蛋白相互作用蛋白1(Huntingtin interacting protein 1,HIP1),AKT/GSK3β信号通路及上皮间充质转化(epithelial-mesenchymal transition,EMT)关键分子蛋白在食管鳞癌中的表达,探讨HIP1表达与AKT/GSK3β信号通路及EMT之间的相关性,为进一步研究HIP1在食管鳞癌中的作用及其机制提供依据。方法:采用免疫组织化学法检测85例食管鳞癌组织及其对应癌旁组织样本中HIP1,p-GSK3β,EMT标志性分子E-cadherin及Vimentin的表达,并统计分析几者在临床病理特征中的表达差异及HIP1与p-GSK3β,E-cadherin,Vimentin表达之间的相关性。采用荧光定量PCR(RT-qPCR)法检测HIP1,GSK3β,E-cadherin及Vimentin基因在食管癌及其癌旁组织中的表达。结果:HIP1,p-GSK3β,E-cadherin及Vimentin在食管鳞癌组织中的阳性表达率分别为91.8%(78/85),82.35%(70/85),15.29%(13/85),80%(68/85),分别与癌旁组织中的表达差异有统计学意义(P<0.001)。HIP1,p-GSK3β,E-cadherin在不同病理分级组间的表达差异有统计学意义(P<0.05),而Vimentin在临床病理特征各组组间的表达差异均无统计学意义(P>0.05)。相关性结果显示,HIP1与p-GSK3β及Vimentin在总的食管鳞癌中的表达均呈显著正相关(P<0.05),HIP1与E-cadherin在总的食管鳞癌中的表达呈显著负相关(P<0.01)。RT-qPCR结果发现,HIP1,GSK3β及Vimentin基因在食管鳞癌中的表达均高于其对应的癌旁组织,而E-cadherin基因在食管鳞癌中的表达低于其对应的癌旁组织。结论:HIP1表达与p-GSK3β,E-cadherin及Vimentin间存在相关性,提示HIP1可能促进食管鳞癌组织中GSK3β及Vimentin的表达,抑制E-cadherin的表达。
Objective: To explore the effect and mechanism of HIP1 in esophageal squamous cell carcinoma(ESCC)carcinogenesis through exam the expression and correlation research of HIP1 with AKT/GSK3β signal pathway and epithelial-mesenchymal transition(EMT) in ESCC. Methods: In this study, HIP1, p-GSK3β, E-cadherin,Vimentin expressions were assessed using immunohistochemistry(IHC) in 85 ESCC and adjacent ESCC tissues and the correlation research of HIP1 with p-GSK3β, E-cadherin, Vimentin were analyzed. RT-qPCR was used to detect the gene expression of HIP1, GSK3β, E-cadherin and Vimentin in ESCC and adjacent ESCC tissues.Results: The expression rate of HIP1, p-GSK3β, E-cadherin, Vimentin in ESCC(91.8%, 82.35%, 15.29%, 80%)showed extremely significance compared with adjacent ESCC tissues(P<0.001). HIP1, p-GSK3β, E-cadherin expression was significantly associated with pathological grades(P<0.05), and there was no significance between Vimentin expression with clinicopathologic features(P>0.05). The correlation research showed that there were significant positive correlations between HIP1 and p-GSK3β, as well as Vimentin(P<0.05), and there was significant negative correlation between HIP1 and E-cadherin(P<0.01). RT-qPCR results showed that compared with adjacent ESCC tissues, HIP1, GSK3β and Vimentin gene were higher expressed in ESCC tissues,but E-cadherin gene was lower expressed in ESCC tissues. Conclusion: HIP1 maybe promote p-GSK3β and Vimentin expression and inhibit E-cadherin expression.
引文
1.Kalchman MA,Koide HB,McCutcheon K,et al.HIP1,a human homologue of S.cerevisiae Sla2p,interacts with membrane-associated huntingtin in the brain[J].Nat Genet,1997,16(1):44-53.
2.Wanker EE,Rovira C,Scherzinger E,et al.HIP-I:a huntingtin interacting protein isolated by the yeast two-hybrid system[J].Hum Mol Genet,1997,6(3):487-495.
3.Wijayatunge R,Holmstrom SR,Foley SB,et al.Deficiency of the Endocytic Protein Hip1 Leads to Decreased Gdpd3 Expression,Low Phosphocholine,and Kypholordosis[J].Mol Cell Biol,2018,38(23).
4.Li D,Chen F,Ding J,et al.Knockdown of HIP1 expression promotes ligand-induced endocytosis of EGFR in Hela cells[J].Oncol Rep,201738(6):3387-3391.
5.Peng L,Yang Q,Xu X,et al.Huntingtin-interacting protein 1-related protein plays a critical role in dendritic development and excitatory synapse formation in hippocampal neurons[J].Front Mol Neurosci2017,10:186.
6.Gottfried I,Ehrlich M,Ashery U.The Sla2p/HIP1/HIP1R family:similar structure,similar function in endocytosis?[J].Biochem Soc Trans,2010,38(Pt 1):187-191.
7.Wilbur JD,Hwang PK,Brodsky FM,et al.Accommodation of structural rearrangements in the huntingtin-interacting protein 1coiled-coil domain[J].Acta Crystallogr D Biol Crystallogr,2010,66(Pt3):314-318.
8.Rice MA,Ishteiwy RA,Magani F,et al.The microRNA-23b/-27b cluster suppresses prostate cancer metastasis via Hunting-interacting protein 1-related[J].Oncogene,2016,35(36):4752-4761.
9.Wang J,Yu M,Guo Q,et al.Prognostic significance of huntingtin interacting protein 1 expression on patients with acute myeloid leukemia[J].Sci Rep,2017,7:45960.
10.Hong M,Kim RN,Song JY,et al.HIP1-ALK,a novel fusion protein identified in lung adenocarcinoma[J].J Thorac Oncol,2014,9(3):419-422.
11.张亚斌,孙盈,孙蕾,等.HIP1在食管癌中的表达及其临床意义[J].现代肿瘤医学,2018,26(21):3401-3404.ZHANG Yabin,SUN Ying,SUN Lei,et al.Expression of HIP1 in esophageal squamous cell carcinoma and its clinical significance[J]Journal of Modern Oncology,2018,26(21):3401-3404.
12.Socinski MA,Crowell R,Hensing TE,et al.Treatment of non-small cell lung cancer,stage IV:ACCP evidence-based clinical practice guidelines(2nd edition)[J].Chest,2007,132(3 Suppl):277S-289S.
13.Azzoli CG,Baker S Jr,Temin S,et al.American Society of Clinical Oncology Clinical Practice Guideline update on chemotherapy for stage IV non-small-cell lung cancer[J].J Clin Oncol,2009,27:6251-6266.
14.Felip E,Gridelli C,Baas P,et al.Metastatic non-small-cell lung cancer:consensus on pathology and molecular tests,first-line,second-line,and third-line therapy:1st ESMO Consensus Conference in Lung Cancer;Lugano 2010[J].Ann Oncol,2011;22:1507-1519.
15.van Klaveren RJ.Lung cancer screening[J].Eur J Cancer,2011,47(Suppl 3):S147-S155.
16.Hsu CY,Lin CH,Jan YH,et al.HIP1 is an early-stage prognostic biomarker of lung adenocarcinoma and suppresses metastasis via Aktmediated EMT[J].Am J Respir Crit Care Med,2016,193(8):869-880.
17.Kao SH,Wang WL,Chen CY,et al.GSK3beta controls epithelialmesenchymal transition and tumor metastasis by CHIP-mediated degradation of Slug[J].Oncogene,2014,33:3172-3182.
18.Scanlon CS,Van Tubergen EA,Inglehart RC,et al.Biomarkers of epithelial-mesenchymal transition in squamous cell carcinoma[J]J Dent Res,2013,92(2):114-121.
19.Turley EA,Veiseh M,Radisky DC,et al.Mechanisms of disease:epithelial-mesenchymal transition--does cellular plasticity fuel neoplastic progression?[J].Nat Clin Pract Oncol,2008,5(5):280-290.
20.Denlinger CE,Ikonomidis JS,R eed CE,et al.Epithelial to mesenchymal transition:the doorway to metastasis in human lung cancers[J].J Thorac Cardiovasc Surg,2010,140(3):505-513.
21.Zheng H,Kang Y.Multilayer control of the EMT master regulators[J]Oncogene,2014,33(14):1755-1763.
22.De Craene B,Berx G.Regulatory networks defining EMT during cancer initiation and progression[J].Nat Rev Cancer,2013,13(2):97-110.
23.Yang J,Weinberg RA.Epithelial-mesenchymal transition:at the crossroads of development and tumor metastasis[J].Dev Cell,200814(6):818-829.