Tollip在结肠癌中的表达及与临床病理参数的关系
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摘要
目的:探讨Tollip在结肠癌中的表达及其与临床病理参数的关系。方法:收集临床结肠癌及癌旁组织标本,采用免疫组织化学、实时荧光定量聚合酶链反应和免疫印迹法分别检测Tollip及相关因子TGF-β1、Smad、E-cadherin、Vimentin的表达。结果:Tollip在结肠癌及癌旁肠黏膜组织上呈弥漫或颗粒状分布于细胞膜与细胞质上,细胞核上无表达。免疫印迹结果显示,结肠癌组Tollip的蛋白表达水平显著高于癌旁组(1.630±0.397 vs 0.651±0.170);RTqPCR结果显示结肠癌组Tollip的mRNA表达水平显著高于癌旁组(3.306±0.528vs0.999±0.005);Tollip的mRNA表达与TGF-β1、Smad2、Smad3的表达呈显著正相关;Tollip的mRNA表达与Vimentin及E-cadherin的表达无相关性。Tollip的mRNA在结肠癌TNM分期中表达差异无统计学意义。结论:结肠癌组织中Tollip的表达升高,可能参与了结肠肿瘤的病理过程,Tollip与TGF-β1/Smad信号通路存在一定的相关性。
Objective :To explore the expression of Tollip and its relationship with clinicopathological parameters in colon cancer. Methods :Surgical specimens were collected from the clinical colon cancer tissues and the paracancerous tissues. The expression of Tollip and its related factors including TGF-β1,Smad,E-cadherin and Vimentin were detected by immunohistochemistry,real-time quantitative polymerase chain reaction(RT-qPCR) and Western blotting. Results :Tollip was diffused or granular in the mucosal tissues of colon cancer and its paracancerous tissues,but was not expressed in the nucleus of the cell. Western blotting results showed that the protein expression level of Tollip in the colon cancer group was higher than that in the paracancerous group(1.630±0.397 vs 0.651±0.170). RTqPCR results showed that the mRNA expression level of Tollip in the colon cancer group(3.306±0.528 vs 0.999±0.005) was higher than that in the paracancerous group. The mRNA expression of Tollip was positively correlated with the expressions of TGF-β1,Smad2 and Smad3;The mRNA expression of Tollip was not correlated with Vimentin and E-cadherin. There was no statistically significant difference in the expression of Tollip mRNA in TNM staging.Conclusion :The expression of Tollip in colon cancer tissues may be involved in the development of colonic tumors,and there is a certain correlation between Tollip and TGF-β1/Smad signaling pathways.
Objective :To explore the expression of Tollip and its relationship with clinicopathological parameters in colon cancer. Methods :Surgical specimens were collected from the clinical colon cancer tissues and the paracancerous tissues. The expression of Tollip and its related factors including TGF-β1,Smad,E-cadherin and Vimentin were detected by immunohistochemistry,real-time quantitative polymerase chain reaction(RT-qPCR) and Western blotting. Results :Tollip was diffused or granular in the mucosal tissues of colon cancer and its paracancerous tissues,but was not expressed in the nucleus of the cell. Western blotting results showed that the protein expression level of Tollip in the colon cancer group was higher than that in the paracancerous group(1.630±0.397 vs 0.651±0.170). RTqPCR results showed that the mRNA expression level of Tollip in the colon cancer group(3.306±0.528 vs 0.999±0.005) was higher than that in the paracancerous group. The mRNA expression of Tollip was positively correlated with the expressions of TGF-β1,Smad2 and Smad3;The mRNA expression of Tollip was not correlated with Vimentin and E-cadherin. There was no statistically significant difference in the expression of Tollip mRNA in TNM staging.Conclusion :The expression of Tollip in colon cancer tissues may be involved in the development of colonic tumors,and there is a certain correlation between Tollip and TGF-β1/Smad signaling pathways.
引文
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[16]van Caam A,Madej W,Garcia de Vinuesa A,et al.TGFβ1-induced SMAD2/3 and SMAD1/5 phosphorylation are both ALK5-kinase-dependent in primary chondrocytes and mediated by TAK1kinase activity[J].Arthritis Res Ther,2017,19(1):112.
[17]Zhu L,Wang L,Luo X,et al.Tollip,an intracellular trafficking protein,is a novel modulator of the transforming growth factor-βsignaling pathway[J].J Biol Chem,2012,287(47):39653-39663.
[18]Pimentel-Nunes P,Gon?alves N,Boal-CarvalhoI,et al.Decreased Toll-interacting protein and peroxisome proliferatoractivated receptorγare associated with increased expression of Toll-like receptors in colon carcinogenesis[J].J Clin Pathol,2012,65(4):302-308.
[19]Draht M X,Smits K M,Tournier B,et al.Promoter CpG island methylation of RET predicts poor prognosis in stage II colorectal cancer patients[J].Mol Oncol,2014,8(3):679-688.
[20]Ahmed M,Lai T H,Zada S,et al.Functional linkage of RKIP to the epithelial to mesenchymal transition and autophagy during the development of prostate cancer[J].Cancers(Basel),2018,10(8).pii:E273.
[21]刘天舟,齐宇.食管鳞状细胞癌组织中Tollip蛋白的表达[J].郑州大学学报医学版,2011,46(2):282-284.
[22]Fang L L,Sun B F,Huang L R,et al.Potent inhibition of miR-34b on migration and invasion in metastatic prostate cancer cells by regulating the TGF-βpathway[J].Int J Mol Sci,2017,18(12)pii:E2762.
[23]Jung B,Staudacher J J,Beauchamp D.Transforming growth factorβsuperfamily signaling in development of colorectal cancer[J].Gastroenterology,2017,152(1):36-52.
[24]Ginnebaugh K R,Ahmad A,Sarkar F H.The therapeutic potential of targeting the epithelial-mesenchymal transition in cancer[J].Expert Opin Ther Targets,2014,18(7):731-745.
[2]徐宁,于振海,王志强,等.Toll样受体4及其负性调控因子Tollip在结肠炎大鼠肠黏膜中的表达[J].第二军医大学报,2012,33(1):32-38.
[3]Xu N,Yu ZH,Yao QS,et al.P PAR-γ and Tollipare associated with toll-like receptors in colitis rats[J].J Immunoassay Immunochem,2013,34(3):219-231.
[4]Wang E L,Qian Z R,Nakasono M,et al.High expression of Toll-like receptor 4/myeloid differentiation factor 88 signals correlates with poor prognosis in colorectal cancer[J].Br J Cancer,2010,102(5):908-915.
[5]Wang CH,Wang PJ,Hsieh YC,et al.Resistin facilitates breast cancer progression via TLR4-mediated induction of mesenchymal phenotypes and stemness properties[J].Oncogene,2018,37(5):589-600.
[6]Parmar N,Chandrakar P,Vishwakarma P,et al.Leishmania donovani Exploits Tollip,a Multitasking Protein,To Impair TLR/IL-1R Signaling for Its Survival in the Host[J].J Immunol,2018,201(3):957-970.
[7]Byun EB,Kim WS,Sung NY,et al.Epigallocatech in-3-Gallate regulates anti-inflammatory action through 67-kDa laminin receptor-mediated Tollip signaling induction in lipopolysaccharidestimulated human intestinal epithelial cells[J].Cell Physiol Biochem,2018,46(5):2072-2081.
[8]Chen K,Yuan R,Zhang Y,et al.Tollip deficiency alters atherosclerosis and steatosis by disrupting lipophagy[J].J Am Heart Assoc,2017,6(4):pii:e004078.
[9]Humbert-Claude M,Duc D,Dwir D,et al.Tollip,an early regulator of the acute inflammatory response in the substantia nigra[J].J Neuroinflammation,2016,13(1):303.
[10]Bermudez-Brito M,Mu?oz-Quezada S,Gomez-Llorente C,et al.Human intestinal dendritic cells decrease cytokine release against Salmonella infection in the presence of Lactobacillus paracasei upon TLR activation[J].PLoS One,2012,7(8):e43197.
[11]Ahmadi A,Najafi M,Farhood B,et al.Transforming growth factor-βsignaling:Tumorigenesis and targeting for cancer therapy[J].J Cell Physiol,2019,234(8):12173-12187.
[12]Colak S,Ten Dijke P.Targeting TGF-βsignaling in cancer[J].Trends Cancer,2017,3(1):56-71.
[13]Neuzillet C,Tijeras-Raballand A,Cohen R,et al.Targeting the TGFβpathway for cancer therapy[J].Pharmacol Ther,2015,147:22-31.
[14]Chen QZ,Li Y,Shao Y,et al.TGF-β1/PTEN/PI3K signaling plays a critical role in the anti-proliferation effect of tetrandrine in human colon cancer cells[J].Int J Oncol,2017,50(3):1011-1021.
[15]Χu C,Lu G,Li Q,et al.Selenium modulates MMP2 expression through the TGFβ1/Smad signalling pathway in human umbilical vein endothelial cells and rabbits following lipid disturbance[J].JTrace Elem Med Biol,2017;42:59-67.
[16]van Caam A,Madej W,Garcia de Vinuesa A,et al.TGFβ1-induced SMAD2/3 and SMAD1/5 phosphorylation are both ALK5-kinase-dependent in primary chondrocytes and mediated by TAK1kinase activity[J].Arthritis Res Ther,2017,19(1):112.
[17]Zhu L,Wang L,Luo X,et al.Tollip,an intracellular trafficking protein,is a novel modulator of the transforming growth factor-βsignaling pathway[J].J Biol Chem,2012,287(47):39653-39663.
[18]Pimentel-Nunes P,Gon?alves N,Boal-CarvalhoI,et al.Decreased Toll-interacting protein and peroxisome proliferatoractivated receptorγare associated with increased expression of Toll-like receptors in colon carcinogenesis[J].J Clin Pathol,2012,65(4):302-308.
[19]Draht M X,Smits K M,Tournier B,et al.Promoter CpG island methylation of RET predicts poor prognosis in stage II colorectal cancer patients[J].Mol Oncol,2014,8(3):679-688.
[20]Ahmed M,Lai T H,Zada S,et al.Functional linkage of RKIP to the epithelial to mesenchymal transition and autophagy during the development of prostate cancer[J].Cancers(Basel),2018,10(8).pii:E273.
[21]刘天舟,齐宇.食管鳞状细胞癌组织中Tollip蛋白的表达[J].郑州大学学报医学版,2011,46(2):282-284.
[22]Fang L L,Sun B F,Huang L R,et al.Potent inhibition of miR-34b on migration and invasion in metastatic prostate cancer cells by regulating the TGF-βpathway[J].Int J Mol Sci,2017,18(12)pii:E2762.
[23]Jung B,Staudacher J J,Beauchamp D.Transforming growth factorβsuperfamily signaling in development of colorectal cancer[J].Gastroenterology,2017,152(1):36-52.
[24]Ginnebaugh K R,Ahmad A,Sarkar F H.The therapeutic potential of targeting the epithelial-mesenchymal transition in cancer[J].Expert Opin Ther Targets,2014,18(7):731-745.