敲低脑内皮细胞粘附分子抑制HNSC细胞的侵袭能力
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摘要
转移是包含头颈部鳞状细胞癌(head-and-neck squamous cell carcinoma, HNSC)在内的多种肿瘤复发和患者死亡的主要原因。目前,关于HNSC转移的网络调控机制仍有待进一步完善,以期降低HNSC死亡率。首先,通过在线数据库GEPIA统计后发现,脑内皮细胞粘附分子(cerebral endothelial cell adhesion molecule, CERCAM)在519例头颈部肿瘤中的相对表达量为4.98,是其在44例正常组织中(相对表达量为2.47)表达量的2.02倍。并且发现CERCAM表达量与头颈部肿瘤患者较差的预后正相关(P=0.015)。其次,在舌癌细胞SCC-9、喉癌细胞HEP2和鼻咽癌细胞CNE2中敲低CERCAM的表达,发现上述3种细胞的侵袭能力均较对照组分别下降93.60%、37.18%和40.63%。最后,生物信息学预测结合Western印迹的结果证实,敲低CERCAM后,上调E-钙黏蛋白(E-cadherin),同时下调锌指E盒结合蛋白(zinc-finger E-box-binding homeobox1, ZEB1)、波形蛋白(vimentin)和Twist相关蛋白1(Twist-related protein 1, TWIST1)。结果证实,CERCAM可能通过调控头颈部肿瘤细胞的上皮间充质转化(epithelial-mesenchymal transition, EMT)标志物来促进该类细胞发生侵袭。
Most cancers including head-and-neck squamous cell carcinoma(HNSC) have high recurrence and mortality because of metastasis. The metastasis regulation networks need further research to attenuate the mortality rate of HNSC. At first, we analyzed the GEPIA databank and found that CERCAM was significantly enhanced in 519 cases of HNSC tissues than that in 44 cases of normal tissues. In addition, high expression of CERCAM was positively correlated to poor prognosis of patients with HNSC. Moreover, shRNAs targeting CERCAM were used to knockdown its expression in SCC-9, HEP2, and CNE2 cells. Comparing with the control group, the invasive ability of SCC-9, HEP2, and CNE2 cells with shRNAs was sharply decreased by 93.60%, 37.18% and 40.63%, respectively. The similar results were found by Western blot assays. Finally, bioinformatics analysis combined with Western blot assays demonstrated that knockdown of CERCAM could evidently enhance expression of E-cadherin, but inhibit expression of ZEB1, vimentin and TWISIT1 in HNSC cells. The data indicated that CERCAM might promote invasion of HNSC cells via EMT.
Most cancers including head-and-neck squamous cell carcinoma(HNSC) have high recurrence and mortality because of metastasis. The metastasis regulation networks need further research to attenuate the mortality rate of HNSC. At first, we analyzed the GEPIA databank and found that CERCAM was significantly enhanced in 519 cases of HNSC tissues than that in 44 cases of normal tissues. In addition, high expression of CERCAM was positively correlated to poor prognosis of patients with HNSC. Moreover, shRNAs targeting CERCAM were used to knockdown its expression in SCC-9, HEP2, and CNE2 cells. Comparing with the control group, the invasive ability of SCC-9, HEP2, and CNE2 cells with shRNAs was sharply decreased by 93.60%, 37.18% and 40.63%, respectively. The similar results were found by Western blot assays. Finally, bioinformatics analysis combined with Western blot assays demonstrated that knockdown of CERCAM could evidently enhance expression of E-cadherin, but inhibit expression of ZEB1, vimentin and TWISIT1 in HNSC cells. The data indicated that CERCAM might promote invasion of HNSC cells via EMT.
引文
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[3] Siegel RL,Miller KD,Jemal A.Cancer statistics,2017[J].CA Cancer J Clin,2017,67(1):7- 30
[4] Kim MM,Califano JA.Molecular pathology of head-and-neck cancer[J].Int J Cancer,2004,112(4):545-553
[5] Chen YP,Wang YQ,Lv JW,et al.Identification and validation of novel microenvironment- based immune molecular subgroups of head and neck squamous cell carcinoma:implications for immunotherapy[J].Ann Oncol,2019,30(1):68-75
[6] Starzyk RM,Rosenow C,Frye J,et al.Cerebral cell adhesion molecule:a novel leukocyte adhesion determinant on blood-brain barrier capillary endothelium[J].J Infect Dis,2000,181 (1):181-187
[7] Tyler RE,Pearce MM,Shaler TA,et al.Unassembled CD147 is an endogenous endoplasmic reticulum-associated degradation substrate[J].Mol Biol Cell,2012,23(24):4668-4678
[8] Tohme S,Simmons RL,Tsung A.Surgery for Cancer:A Trigger for Metastases[J].Cancer Res,2017,77(7):1548-1552
[9] Peinado H,Zhang H,Matei IR,et al.Pre-metastatic niches:organ-specific homes for metastases[J].Nat Rev Cancer,2017,17(5):302-317
[10] Zhang L,Liang Y,Li S,et al.The interplay of circulating tumor DNA and chromatin modification,therapeutic resistance,and metastasis[J].Mol Cancer,2019,18(1):36
[11] Pectasides E,Rampias T,Sasaki C,et al.Markers of epithelial to mesenchymal transition in association with survival in head and neck squamous cell carcinoma (HNSCC)[J].PLoS One,2014,9(4):e94273
[12] Goossens S,Vandamme N,Van Vlierberghe P,et al.EMT transcription factors in cancer development re-evaluated:Beyond EMT and MET[J].Biochim Biophys Acta Rev Cancer,2017,1868(2):584-591
[13] Milsom CC,Yu JL,Mackman N,et al.Tissue factor regulation by epidermal growth factor receptor and epithelial-to-mesenchymal transitions:effect on tumor initiation and angiogenesis [J].Cancer Res,2008,68(24):10068-10076
[14] Zhang L,Sun J,Wang B,et al.MicroRNA-10b triggers the epithelial-mesenchymal transition (EMT) of laryngeal carcinoma Hep-2 cells by directly targeting the E-cadherin[J].Appl Biochem Biotechnol,2015,176(1):33-44
[15] Mandal M,Myers JN,Lippman SM,et al.Epithelial to mesenchymal transition in head and neck squamous carcinoma:association of Src activation with E-cadherin down-regulation,vimentin expression,and aggressive tumor features[J].Cancer,2008,112(9):2088-2100
[16] Zeeshan R,Mutahir Z.Cancer metastasis - tricks of the trade[J].Bosn J Basic Med Sci,2017,17(3):172-182
[17] Fidler IJ,Kripke ML.The challenge of targeting metastasis[J].Cancer Metastasis Rev,2015,34(4):635-641
[18] Cappellesso R,Marioni G,Crescenzi M,et al.The prognostic role of the epithelial- mesenchymal transition markers E-cadherin and Slug in laryngeal squamous cell carcinoma[J].Histopathology,2015,67(4):491-500
[19] Wang W,Yi M,Zhang R,et al.Vimentin is a crucial target for anti-metastasis therapy of nasopharyngeal carcinoma[J].Mol Cell Biochem,2018,438(1-2):47-57
[20] da Silva SD,Alaoui-Jamali MA,Soares FA,et al.TWIST1 is a molecular marker for a poor prognosis in oral cancer and represents a potential therapeutic target[J].Cancer,2014,120(3):352-362