食管鳞状细胞癌中单羧酸转运蛋白4的表达及对EC109细胞增殖、迁移和侵袭能力的影响
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摘要
目的:探讨食管鳞状细胞癌(食管鳞癌)中单羧酸转运蛋白4(MCT4)的表达及对EC109细胞增殖、迁移和侵袭能力的影响。方法:免疫组化法检测60例食管鳞癌和19例正常食管黏膜组织中MCT4蛋白的表达。Western blot法检测正常食管上皮细胞Het1-A和食管鳞癌细胞EC109、EC9706中MCT4蛋白的表达。将靶向MCT4的siRNA转染至EC109细胞中,Western blot法检测MCT4蛋白的表达,采用乳酸试剂盒检测胞外乳酸含量,采用CCK-8法、划痕实验和Transwell技术检测细胞增殖率、迁移率和侵袭率;以未转染细胞和转染阴性对照siRNA的细胞作对照。结果:食管鳞癌组织中MCT4高表达率高于正常组织(70. 0%vs 26. 3%,P <0. 001),低分化癌组织中的高表达率高于高中分化癌组织(P=0. 037)。EC9706、EC109细胞中MCT4蛋白表达水平高于Het1-A(P <0. 05)。与对照组比较,MCT4 siRNA转染的EC109细胞胞外乳酸含量降低,细胞增殖率、迁移率和侵袭率均降低(P <0. 001)。结论:下调MCT4的表达能够抑制食管鳞癌细胞的恶性生物学行为。
Aim: To investigate the expression of monocarboxylate transporter 4(MCT4) and its effects on the proliferation,migration,and invasion capacity of esophageal squamous cell carcinoma(ESCC). Methods: Immunohistochemistry was used to detect the MCT4 protein expression in 60 cases of ESCC and 19 cases of adjacent normal tissue. The expression of MCT4 in ESCC cell lines(EC109,EC9706) and normal esophageal epithelium cell(Het1-A) was detected by Western blot. The siRNA targeted at MCT4(MCT4 siRNA) was transfected into EC109 cells,and the MCT4 protein expression was detected by Western blot,the extracellular lactic acid content was detected by lactic acid kit,the proliferation,migration and invasion were detected by CCK-8 method,wound healing test and Transwell technique,respectively. The cells without transfection or transfected with negative siRNA were the controls. Results: The high expression rate of MCT4 in ESCC tissue was higher than that in adjacent normal tissue(70. 0% vs 26. 3%,P < 0. 001),which was correlated with pathological differentiation(P = 0. 037). The expression of MCT4 protein in EC109 and EC9706 cells was higher than that in Het1-A cells(P < 0. 05). Compared with the controls,the extracellular lactic acid content decreased,and cell proliferation,migration,and invasion abilities of the cells transfected with MCT4 siRNA decreased(P < 0. 001). Conclusion: Down-regulation of MCT4 can inhibit the malignant biological behavior of ESCC cells.
Aim: To investigate the expression of monocarboxylate transporter 4(MCT4) and its effects on the proliferation,migration,and invasion capacity of esophageal squamous cell carcinoma(ESCC). Methods: Immunohistochemistry was used to detect the MCT4 protein expression in 60 cases of ESCC and 19 cases of adjacent normal tissue. The expression of MCT4 in ESCC cell lines(EC109,EC9706) and normal esophageal epithelium cell(Het1-A) was detected by Western blot. The siRNA targeted at MCT4(MCT4 siRNA) was transfected into EC109 cells,and the MCT4 protein expression was detected by Western blot,the extracellular lactic acid content was detected by lactic acid kit,the proliferation,migration and invasion were detected by CCK-8 method,wound healing test and Transwell technique,respectively. The cells without transfection or transfected with negative siRNA were the controls. Results: The high expression rate of MCT4 in ESCC tissue was higher than that in adjacent normal tissue(70. 0% vs 26. 3%,P < 0. 001),which was correlated with pathological differentiation(P = 0. 037). The expression of MCT4 protein in EC109 and EC9706 cells was higher than that in Het1-A cells(P < 0. 05). Compared with the controls,the extracellular lactic acid content decreased,and cell proliferation,migration,and invasion abilities of the cells transfected with MCT4 siRNA decreased(P < 0. 001). Conclusion: Down-regulation of MCT4 can inhibit the malignant biological behavior of ESCC cells.
引文
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[18]KIM HK,LEE IK,BANG HE,et al.MCT4 expression is a potential therapeutic target in colorectal cancer with peritoneal carcinomatosis[J].Mol Cancer Ther,2018,17(4):838
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[2]ARNOLD M,SOERJOMATARAM I,FERLAY J,et al.Global incidence of oesophageal cancer by histological subtype in 2012[J].Gut,2015,64(3):381
[3]KEKUDA R,MANOHARAN P,BASELER WA.Monocarboxylate 4 mediated butyrate transport in a rat intestinal epithelial cell line[J].Dig Dis Sci,2013,58(3):660
[4]HALESTRAP AP.The SLC16 gene family-structure,role and regulation in health and disease[J].Mol Aspects Med,2013,34(2/3,SI):337
[5]HUHTA H,HELMINEN O,PALOMKI S,et al.Intratumoral lactate metabolism in barrett's esophagus and adenocarcinoma[J].Oncotarget,2017,8(14):22894
[6]LEE JY,LEE IK,CHANG WJ,et al.MCT4 as a potential therapeutic target for metastatic gastric cancer with peritoneal carcinomatosis[J].Oncotarget,2016,7(28):43492
[7]KIM HK,LEE IK,BANG H,et al.MCT4 expression is a potential therapeutic target in colorectal cancer with peritoneal carcinomatosis[J].Mol Cancer Ther,2018,17(4):838
[8]郭振江,刘宗文,郑瑞锋,等.沉默单羧酸转运体4对宫颈癌Hela、Siha细胞增殖、侵袭迁移及凋亡的影响[J].郑州大学学报(医学版),2018,53(3):354
[9]JOHNSON LL,PAVLOVSKY AG,JOHNSON AR,et al.Arationalization of the acidic p H dependence for stromelysin-1(matrix metalloproteinase-3)catalysis and inhibition[J].J Biol Chem,2000,275(15):11026
[10]BAUMANN F,LEUKEL P,DOERFELT A,et al.Lactate promotes glioma migration by TGF-beta2-dependent regulation of matrix metalloproteinase-2[J].Neuro Oncol,2009,11(4):368
[11]张宝月,瞿全新,顾安康.CD147、MCT1及MCT4在宫颈病变中的表达及临床意义[J].国际妇产科学杂志,2015,42(2):194
[12]VOSS DM,SPINA R,CARTER DL,et al.Disruption of the monocarboxylate transporter-4-basigin interaction inhibits the hypoxic response,proliferation,and tumor progression[J].Sci Rep,2017,7(1):4292
[13]YU J,SUN JT,FAN YS,et al.Exposure to Pb and Cd alters MCT4/CD147 expression and MCT4/CD147-dependent lactate transport in mice Sertoli cells cultured in vitro[J].Toxicol In Vitro,2019,56:30
[14]PARK SJ,SMITH CP,WILBUR RR,et al.An overview of MCT1 and MCT4 in GBM:small molecule transporters with large implications[J].Am J Cancer Res,2018,8(10):1967
[15]ZHU J,WU YN,ZHANG W,et al.Monocarboxylate transporter 4 facilitates cell proliferation and migration and is associated with poor prognosis in oral squamous cell carcinoma patients[J].PLo S One,2014,9(1):e87904
[16]GAO HJ,ZHAO MC,ZHANG YJ,et al.Monocarboxylate transporter 4 predicts poor prognosis in hepatocellular carcinoma and is associated with cell proliferation and migration[J].J Cancer Res Clin Oncol,2015,141(7):1151
[17]TODENHFER T,SEILER R,STEWART C,et al.Selective inhibition of the lactate transporter MCT4 reduces growth of invasive bladder cancer[J].Mol Cancer Ther,2018,17(12):2746
[18]KIM HK,LEE IK,BANG HE,et al.MCT4 expression is a potential therapeutic target in colorectal cancer with peritoneal carcinomatosis[J].Mol Cancer Ther,2018,17(4):838
[19]CHOI SY,ETTINGER SL,LIN D,et al.Targeting MCT4 to reduce lactic acid secretion and glycolysis for treatment of neuroendocrine prostate cancer[J].Cancer Med,2018,7(7):3385