Sox1过表达对口腔鳞癌细胞增殖、侵袭及Wnt通路的影响
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摘要
目的研究性别决定区域Y盒1(Sox1)过表达对口腔鳞癌细胞增殖、迁移以及Wnt通路的影响。方法荧光实时定量聚合酶链反应(qRT-PCR)法检测人正常口腔组织细胞系及口腔鳞癌细胞系HN4中Sox1 mRNA表达。采用Lipofectamin 2000试剂进行转染,分为三组:空白组、Sox1 NC组、Sox1 mimics组,每孔别按照不添加、0. 7μl 20μmol/L的p CMV6空载体、0. 7μl 20μmol/L的p CMV6-Sox1的水平加入相应试剂。检测Sox1 mRNA和蛋白水平,通过MTT试剂盒检测细胞增殖,Transwell法检测细胞侵袭能力,免疫印迹(Western blot)法检测Wnt通路相关蛋白表达水平。结果与正常细胞相比,口腔鳞癌细胞中Sox1 mRNA、Sox1蛋白表达水平显著降低(P <0. 05);空白组与Sox1 NC组Sox1 mRNA、蛋白表达差异无统计学意义(P> 0. 05),与空白组、Sox1 NC组相比,Sox1 mimic组Sox1 mRNA、蛋白表达显著升高(P <0. 05); MTT试验显示,Sox1 NC组与空白组相比,0 h、24 h、48 h、72 h、96 h,OD值差异无统计学意义(P>0. 05),与Sox1 NC组和空白组相比,Sox1 mimic组转染后72 h后OD值显著降低(P <0. 05); Western blot检测结果显示,空白组与Sox1 NC组相比Wnt、GSK-3β、β-Catenin、cyclin D1蛋白表达无显著差异(P> 0. 05),与空白组、Sox1 NC相比,Sox1 mimic组Wnt、β-Catenin、cyclin D1蛋白表达明显下降(P <0. 05),GSK-3β表达量上升(P <0. 05)。结论口腔鳞癌组织中Sox1低表达,Sox1过表达通过Wnt通路降低口腔鳞癌细胞增殖、侵袭能力。
Objective To study the effects of sex determination region Y-Box 1( Sox1) overexpression on the proliferation,migration,and Wnt pathway of oral squamous cell carcinoma cells. Methods The expression of Sox1 mRNA in human normal oral tissue cell line and oral squamous cell carcinoma cell line HN4 was detected by real-time fluorescence quantitative polymerase chain reaction( qRT-PCR). Transfection was performed using Lipofectamin 2000 reagent and divided into three groups: blank group,Sox1 NC group,and Sox1 mimics group. Add the corresponding reagent to the level of 0. 7 μl 20 μmol/L pCMV6 empty vector and 0. 7 μl 20 μmol/L pCMV6-Sox1 without adding. The levels of Sox1 mRNA and protein were detected,cell proliferation was detected by MTT kit,Transwell assay was used to detect cell invasion ability,Western blot was used to detect the expression of Wnt pathway-related protein. Results Compared with normal cells,the expression levels of Sox1 mRNA and Sox1 protein in oral squamous cell carcinoma cells were significantly decreased( P < 0. 05); there was no significant change in the expressions of Sox1 mRNA and protein of the blank group and the Sox1 NC group( P > 0. 05),compared with blank group and Sox1 NC group,the expressions of Sox1 mRNA and protein in Sox1 mimic group were significantly increased( P < 0. 05); MTT test showed that there was no significant difference in OD value between Sox1 NC group and blank group at each time point( P > 0. 05),compared with Sox1 NC group and blank group,the OD value of Sox1 mimic group decreased significantly at 72 h after transfection( P < 0. 05); Western blot results showed that there was no significant difference in Wnt,GSK-3β,β-Catenin,cyclinD1 protein expression between the blank group with the Sox1 NC group( P > 0. 05),compared with blank group and Sox1 NC,the expressions of Wnt,β-Catenin and cyclinD1 protein in Sox1 mimic group decreased significantly( P < 0. 05),the expression of GSK-3β increased( P < 0. 05). Conclusion Sox1 expression in oral squamous cell carcinoma is low,and overexpression of Sox1 reduces the proliferation and invasion ability of oral squamous cell carcinoma cells through Wnt pathway.
Objective To study the effects of sex determination region Y-Box 1( Sox1) overexpression on the proliferation,migration,and Wnt pathway of oral squamous cell carcinoma cells. Methods The expression of Sox1 mRNA in human normal oral tissue cell line and oral squamous cell carcinoma cell line HN4 was detected by real-time fluorescence quantitative polymerase chain reaction( qRT-PCR). Transfection was performed using Lipofectamin 2000 reagent and divided into three groups: blank group,Sox1 NC group,and Sox1 mimics group. Add the corresponding reagent to the level of 0. 7 μl 20 μmol/L pCMV6 empty vector and 0. 7 μl 20 μmol/L pCMV6-Sox1 without adding. The levels of Sox1 mRNA and protein were detected,cell proliferation was detected by MTT kit,Transwell assay was used to detect cell invasion ability,Western blot was used to detect the expression of Wnt pathway-related protein. Results Compared with normal cells,the expression levels of Sox1 mRNA and Sox1 protein in oral squamous cell carcinoma cells were significantly decreased( P < 0. 05); there was no significant change in the expressions of Sox1 mRNA and protein of the blank group and the Sox1 NC group( P > 0. 05),compared with blank group and Sox1 NC group,the expressions of Sox1 mRNA and protein in Sox1 mimic group were significantly increased( P < 0. 05); MTT test showed that there was no significant difference in OD value between Sox1 NC group and blank group at each time point( P > 0. 05),compared with Sox1 NC group and blank group,the OD value of Sox1 mimic group decreased significantly at 72 h after transfection( P < 0. 05); Western blot results showed that there was no significant difference in Wnt,GSK-3β,β-Catenin,cyclinD1 protein expression between the blank group with the Sox1 NC group( P > 0. 05),compared with blank group and Sox1 NC,the expressions of Wnt,β-Catenin and cyclinD1 protein in Sox1 mimic group decreased significantly( P < 0. 05),the expression of GSK-3β increased( P < 0. 05). Conclusion Sox1 expression in oral squamous cell carcinoma is low,and overexpression of Sox1 reduces the proliferation and invasion ability of oral squamous cell carcinoma cells through Wnt pathway.
引文
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[11]Nie D,Wang Z,Zhang Y,et al.Fat-1 gene inhibits human oral squamous carcinoma cell proliferation through downregulation of-catenin signaling pathways[J].Exp Ther Med,2016,11(1):191-196.
[12]Chen H,Yang H,Pan L,et al.The molecular mechanisms of XBP-1gene silencing on IRE1±-TRAF2-ASK1-JNK pathways in oral squamous cell carcinoma under endoplasmic reticulum stress[J].Biomed Pharmacother,2016,77(1):108-113.
[13]李宁,周素贞,李素云.肺癌细胞中Sox1调控ECT2的表达[J].中国医药科学,2015,5(15):23-26.
[14]程凯,文娟娟,周晓蝶,等.乳腺癌组织中Sox1O的表达及其临床意义[J].临床与实验病理学杂志,2016,32(1):9-12.
[15]Song L,Liu D,He J,et al.Sox1 inhibits breast cancer cell growth and invasion through suppressing the Wnt/2-catenin signaling pathway[J].Apmis,2016,124(7):547-555.
[16]Yang P,Qiu Z,Jiang Y,et al.Silencing of c ZNF292 circular RNAsuppresses human glioma tube formation via the Wnt/-catenin signaling pathway[J].Oncotarget,2016,7(39):63449-63455.
[17]Duffy DJ,Krstic A,Schwarzl T,et al.Wnt signalling is a bi-directional vulnerability of cancer cells[J].Oncotarget,2016,7(37):60310-60331.
[18]Su SF,de Castro Abreu AL,Chihara Y,et al.A panel of three markers hyper-and hypomethylated in urine sediments accurately predicts bladder cancer recurrence[J].Clin Cancer Res,2014,20(7):1978-1989.
[19]Tsao CM,Yan MD,Shih YL,et al.SOX1 functions as a tumor suppressor by antagonizing the WNT/-catenin signaling pathway in hepatocellular carcinoma[J].Hepatology,2012,56(6):2277-2287.
[2]陈芬,陈林林.口腔疣状癌、口腔鳞癌与癌基因关系的研究进展[J].实用临床医学,2016,17(1):92-93.
[3]韩影,苏彤,林璐,等.口腔癌对侧颈淋巴结转移的临床研究[J].口腔医学研究,2016,32(2):154-157.
[4]Huang J,Tan ZR,Yu J,et al.DNA hypermethylated status and gene expression of PAX1/Sox1 in patients with colorectal carcinoma[J].Onco Targets Ther,2017,10(1):4739-4751.
[5]Song L,Liu D,He J,et al.Sox1 inhibits breast cancer cell growth and invasion through suppressing the Wnt/2-catenin signaling pathway[J].Apmis,2016,124(7):547-555.
[6]俞力夫,代晓明,李逸松.mRNA差异表达与口腔鳞状细胞癌相关研究进展[J].口腔医学研究,2016,32(1):100-104.
[7]毕也,周童,张泽兵.影响口腔鳞状细胞癌淋巴结转移及预后生物指标的相关研究进展[J].中国实验诊断学,2017,21(8):1458-1461.
[8]周昱川,王雪阳,陈青立,等.Hippo信号通路中TAZ、P-TAZ、Mst1/2与P-Mst1/2在口腔鳞状细胞癌中的表达及意义[J].肿瘤学杂志,2017,23(3):199-204.
[9]邱峰,王潇悦,赵军方,等.亚砷酸钠调控Wnt信号通路抑制口腔鳞状细胞癌增殖及促进凋亡的机制研究[J].中国生化药物杂志,2017,30(4):12-16.
[10]王雪阳,周昱川,陈青立,等.Hippo信号通路相关蛋白YAP,Mst1,Lats 1/2在口腔鳞癌中的表达及临床意义[J].口腔医学研究,2017,33(4):400-403.
[11]Nie D,Wang Z,Zhang Y,et al.Fat-1 gene inhibits human oral squamous carcinoma cell proliferation through downregulation of-catenin signaling pathways[J].Exp Ther Med,2016,11(1):191-196.
[12]Chen H,Yang H,Pan L,et al.The molecular mechanisms of XBP-1gene silencing on IRE1±-TRAF2-ASK1-JNK pathways in oral squamous cell carcinoma under endoplasmic reticulum stress[J].Biomed Pharmacother,2016,77(1):108-113.
[13]李宁,周素贞,李素云.肺癌细胞中Sox1调控ECT2的表达[J].中国医药科学,2015,5(15):23-26.
[14]程凯,文娟娟,周晓蝶,等.乳腺癌组织中Sox1O的表达及其临床意义[J].临床与实验病理学杂志,2016,32(1):9-12.
[15]Song L,Liu D,He J,et al.Sox1 inhibits breast cancer cell growth and invasion through suppressing the Wnt/2-catenin signaling pathway[J].Apmis,2016,124(7):547-555.
[16]Yang P,Qiu Z,Jiang Y,et al.Silencing of c ZNF292 circular RNAsuppresses human glioma tube formation via the Wnt/-catenin signaling pathway[J].Oncotarget,2016,7(39):63449-63455.
[17]Duffy DJ,Krstic A,Schwarzl T,et al.Wnt signalling is a bi-directional vulnerability of cancer cells[J].Oncotarget,2016,7(37):60310-60331.
[18]Su SF,de Castro Abreu AL,Chihara Y,et al.A panel of three markers hyper-and hypomethylated in urine sediments accurately predicts bladder cancer recurrence[J].Clin Cancer Res,2014,20(7):1978-1989.
[19]Tsao CM,Yan MD,Shih YL,et al.SOX1 functions as a tumor suppressor by antagonizing the WNT/-catenin signaling pathway in hepatocellular carcinoma[J].Hepatology,2012,56(6):2277-2287.