同源形成素样蛋白2基因对口腔鳞状细胞癌侵袭、迁移及核因子-κB信号的影响
|
摘要
目的:探讨下调同源形成素样蛋白2(formin-like 2,FMNL2)基因表达对口腔鳞状细胞癌侵袭、迁移及核因子-κB(nuclear factor-κB,NF-κB)信号的影响。方法:FMNL2的特异性小干扰RNA(small interfering RNA,siRNA)(si-FMNL2组)及阴性对照siRNA(NC组)转染人口腔鳞状细胞癌CAL-27细胞,二硫代氨基甲酸吡咯烷(pyrro1idinedithiocarbamate,PDTC)作为NF-κB信号抑制剂,处理48h,Western blot检测FMNL2、E-钙粘蛋白(E-cadherin)、波形蛋白(Vimentin)、NF-κB p65和IκBα蛋白表达;Transwell小室检测细胞侵袭和迁移能力。结果:与NC组比较,si-FMNL2组和PDTC组细胞侵袭和迁移能力均明显降低,E-cadherin蛋白表达明显升高,Vimentin、NF-κB p65和IκBα蛋白表达明显降低(P<0.05)。与PDTC组比较,PDTC+si-FMNL2组细胞侵袭和迁移能力均明显降低,E-cadherin蛋白表达明显升高,Vimentin蛋白表达明显降低(P<0.05)。结论:下调FMNL2基因表达可通过抑制NF-κB信号通路逆转上皮-间充质转化(epithelial-mesenchymal transition,EMT),从而抑制口腔鳞状细胞癌侵袭和迁移能力。
Objective:To investigate the effect of down-regulation of FMNL2 gene expression on invasion,migration,and NF-κB signal in oral squamous cell carcinoma(OSCC).Methods:FMNL2specific siRNA(si-FMNL2)and negative control siRNA(NC)were transfected into human oral squamous cell carcinoma cells CAL-27.PDTC was set as NF-κB signal inhibitor.After the cells were treated for 48 h,western blotting was used to detect FMNL2,E-cadherin,Vimentin,NF-κBp65,and IκBαprotein expression.Results:Compared with the blank group,the cell invasion and migration ability in si-FMNL2 group and PDTC group decreased obviously,the expression of E-cadherin protein increased obviously,and the expression of Vimentin,NF-κBp65,and IκBαprotein decreased significantly(P<0.05).Compared with the PDTC group,the cell invasion and migration ability in PDTC+si-FMNL2 group decreased obviously,the expression of E-cadherin protein increased obviously,and the expression of Vimentin protein decreased significantly(P<0.05).Conclusion:Downregulation of FMNL2 gene expression can reverse EMT by inhibiting NF-κB signaling pathway,thus inhibiting the invasion and migration of oral squamous cell carcinoma.
Objective:To investigate the effect of down-regulation of FMNL2 gene expression on invasion,migration,and NF-κB signal in oral squamous cell carcinoma(OSCC).Methods:FMNL2specific siRNA(si-FMNL2)and negative control siRNA(NC)were transfected into human oral squamous cell carcinoma cells CAL-27.PDTC was set as NF-κB signal inhibitor.After the cells were treated for 48 h,western blotting was used to detect FMNL2,E-cadherin,Vimentin,NF-κBp65,and IκBαprotein expression.Results:Compared with the blank group,the cell invasion and migration ability in si-FMNL2 group and PDTC group decreased obviously,the expression of E-cadherin protein increased obviously,and the expression of Vimentin,NF-κBp65,and IκBαprotein decreased significantly(P<0.05).Compared with the PDTC group,the cell invasion and migration ability in PDTC+si-FMNL2 group decreased obviously,the expression of E-cadherin protein increased obviously,and the expression of Vimentin protein decreased significantly(P<0.05).Conclusion:Downregulation of FMNL2 gene expression can reverse EMT by inhibiting NF-κB signaling pathway,thus inhibiting the invasion and migration of oral squamous cell carcinoma.
引文
[1] Yu XD,Yang J,Zhang WL,et al.Resveratrol inhibits oral squamous cell carcinoma through induction of apoptosis and G2/M phase cell cycle arrest[J].Tumor Biology,2016,37(3):2871-2877.
[2] Yan G,Wang X,Yang M,et al.Long non-coding RNA TUG1promotes progression of oral squamous cell carcinoma through upregulating FMNL2by sponging miR-219[J].Am J Cancer Res,2017,7(9):1899-1912.
[3] Ji C,Liu H,Yin Q,et al.miR-93enhances hepatocellular carcinoma invasion and metastasis by EMT via targeting PDCD4[J].Biotechnol Lett,2017,39(11):1621-1629.
[4] Zhou W,Wang Q,Xu Y,et al.RMP promotes epithelialmesenchymal transition through NF-κB/CSN2/Snail pathway in hepatocellular carcinoma[J].Oncotarget,2017,8(25):40373-40388.
[5]刘勇,阮思蓓,吕沐瀚,等.转移相关基因FMNL2参与胃癌侵袭转移的相关性分析[J].临床与实验病理学杂志,2018,34(4),439-441.
[6] Li Y,Zhu X,Zeng Y,et al.FMNL2enhances invasion of colorectal carcinoma by inducing epithelial-mesenchymal transition[J].Mol Cancer Res,2010,8(12):1579-1590.
[7]潘斌,邓志海,吴友科,等.沉默URG11基因对前列腺癌LNCaP细胞增殖、迁移与侵袭的影响[J].中国病理生理杂志,2016,32(4):658-664.
[8]朱曦龄,黎功.FMNL2基因对大肠癌细胞SW620增殖、侵袭及转移的影响[J].山东医药,2017,57(24):34-37.
[9]Wang L,Zhao Z,Feng W,et al.Long non-coding RNA TUG1promotes colorectal cancer metastasis via EMT pathway[J].Oncotarget,2016,7(32):51713-51719.
[10] Zong H,Yin B,Zhou H,et al.Inhibition of mTOR pathway attenuates migration and invasion of gallbladder cancer via EMT inhibition[J].Mol Biol Rep,2014,41(7):4507-4512.
[11] Jing Z,Chen XY,Huang KJ,et al.Expression of FoxM1and the EMT-associated protein E-cadherin in gastric cancer and its clinical significance[J].Oncol Lett,2016,12(4):2445-2450.
[12] Hong KO,Lee JI,Hong SP,et al.Thymosinβ4induces proliferation,invasion,and epithelial-to-mesenchymal transition of oral squamous cell carcinoma[J].Amino Acids,2016,48(1):117-127.
[13]陶明珠,张强,周铁军,等.FMNL2基因小干扰RNA质粒转染人宫颈癌Hela、Siha细胞株Vimentin及E-cadherin的表达观察[J].山东医药,2017,5(19):5-8.
[14] Song W,Wang L,Lei Z,et al.ZNF143enhances metastasis of gastric cancer by promoting the process of EMT through PI3K/AKT signaling pathway[J].Tumour Biol,2016,37(9):12813-12821.
[15]杨芳,陈明月,胡英英,等.PPARγ在脂多糖刺激牙周膜细胞中调控NF-κB信号通路的作用研究[J].口腔医学研究,2017,33(7):698-702.
[16] Liang F,Zhang S,Wang B,et al.Overexpression of integrin-linked kinase(ILK)promotes glioma cell invasion and migration and down-regulates E-cadherin via the NF-κB pathway[J].J Mol Histol,2014,45(2):141-151.
[17] Su X,Wang J,Chen W,et al.Overexpression of TRIM14promotes tongue squamous cell carcinoma aggressiveness by activating the NF-κB signaling pathway[J].Oncotarget,2016,7(9):9939-9950.
[18] Yang SS,Li XM,Yang M,et al.FMNL2destabilises COMMD10to activate NF-κB pathway in invasion and metastasis of colorectal cancer[J].Br J Cancer,2017,117(8):1164-1175.
[2] Yan G,Wang X,Yang M,et al.Long non-coding RNA TUG1promotes progression of oral squamous cell carcinoma through upregulating FMNL2by sponging miR-219[J].Am J Cancer Res,2017,7(9):1899-1912.
[3] Ji C,Liu H,Yin Q,et al.miR-93enhances hepatocellular carcinoma invasion and metastasis by EMT via targeting PDCD4[J].Biotechnol Lett,2017,39(11):1621-1629.
[4] Zhou W,Wang Q,Xu Y,et al.RMP promotes epithelialmesenchymal transition through NF-κB/CSN2/Snail pathway in hepatocellular carcinoma[J].Oncotarget,2017,8(25):40373-40388.
[5]刘勇,阮思蓓,吕沐瀚,等.转移相关基因FMNL2参与胃癌侵袭转移的相关性分析[J].临床与实验病理学杂志,2018,34(4),439-441.
[6] Li Y,Zhu X,Zeng Y,et al.FMNL2enhances invasion of colorectal carcinoma by inducing epithelial-mesenchymal transition[J].Mol Cancer Res,2010,8(12):1579-1590.
[7]潘斌,邓志海,吴友科,等.沉默URG11基因对前列腺癌LNCaP细胞增殖、迁移与侵袭的影响[J].中国病理生理杂志,2016,32(4):658-664.
[8]朱曦龄,黎功.FMNL2基因对大肠癌细胞SW620增殖、侵袭及转移的影响[J].山东医药,2017,57(24):34-37.
[9]Wang L,Zhao Z,Feng W,et al.Long non-coding RNA TUG1promotes colorectal cancer metastasis via EMT pathway[J].Oncotarget,2016,7(32):51713-51719.
[10] Zong H,Yin B,Zhou H,et al.Inhibition of mTOR pathway attenuates migration and invasion of gallbladder cancer via EMT inhibition[J].Mol Biol Rep,2014,41(7):4507-4512.
[11] Jing Z,Chen XY,Huang KJ,et al.Expression of FoxM1and the EMT-associated protein E-cadherin in gastric cancer and its clinical significance[J].Oncol Lett,2016,12(4):2445-2450.
[12] Hong KO,Lee JI,Hong SP,et al.Thymosinβ4induces proliferation,invasion,and epithelial-to-mesenchymal transition of oral squamous cell carcinoma[J].Amino Acids,2016,48(1):117-127.
[13]陶明珠,张强,周铁军,等.FMNL2基因小干扰RNA质粒转染人宫颈癌Hela、Siha细胞株Vimentin及E-cadherin的表达观察[J].山东医药,2017,5(19):5-8.
[14] Song W,Wang L,Lei Z,et al.ZNF143enhances metastasis of gastric cancer by promoting the process of EMT through PI3K/AKT signaling pathway[J].Tumour Biol,2016,37(9):12813-12821.
[15]杨芳,陈明月,胡英英,等.PPARγ在脂多糖刺激牙周膜细胞中调控NF-κB信号通路的作用研究[J].口腔医学研究,2017,33(7):698-702.
[16] Liang F,Zhang S,Wang B,et al.Overexpression of integrin-linked kinase(ILK)promotes glioma cell invasion and migration and down-regulates E-cadherin via the NF-κB pathway[J].J Mol Histol,2014,45(2):141-151.
[17] Su X,Wang J,Chen W,et al.Overexpression of TRIM14promotes tongue squamous cell carcinoma aggressiveness by activating the NF-κB signaling pathway[J].Oncotarget,2016,7(9):9939-9950.
[18] Yang SS,Li XM,Yang M,et al.FMNL2destabilises COMMD10to activate NF-κB pathway in invasion and metastasis of colorectal cancer[J].Br J Cancer,2017,117(8):1164-1175.