外源性拮抗神经纤毛蛋白1对人舌癌细胞SCC9细胞迁移及分化的影响

›› 2015, Vol. 6 ›› Issue (4): 198-210.

外源性拮抗神经纤毛蛋白1对人舌癌细胞SCC9细胞迁移及分化的影响

查霖¹,吴煜农²,郑灏¹,赵宏惠¹,安宁¹,宋晓萌²,唐子春³

1. 铜陵市人民医院口腔科
2. 南京医科大学附属口腔医院
3. 安徽省铜陵市人民医院口腔科

收稿日期:2015-06-23 修回日期:2015-08-13 出版日期:2015-12-25 发布日期:2015-12-30
通讯作者: 唐子春 E-mail:tzc9439@126.com
基金资助:
安徽省铜陵市卫生科研项目（2014-26）

Exogenous inhibition of Neuropilin-1 effects on the migration and the differentiation of human tongue squamous cell carcinoma cell line SCC9

Received:2015-06-23 Revised:2015-08-13 Online:2015-12-25 Published:2015-12-30

摘要/Abstract

摘要： 目的利用小分子肽ATWLPPR(A7R)对人舌癌细胞SCC9进行靶向拮抗其神经纤毛蛋白1(Neuropilin-1,NRP-1)，研究其对人舌癌细胞SCC9迁移、分化的影响。方法通过免疫印迹实验（Western Blot，WB）检测NRP-1蛋白在舌癌组织及人舌癌细胞SCC9中的表达情况，利用划痕实验及Transwell检测小分子肽A7R对SCC9细胞迁移的影响，以及利用实时定量荧光PCR（Real-Time RT-PCR）检测相关上皮、间充质标记蛋白的mRNA表达变化，研究小分子肽A7R对SCC9细胞分化的影响。结果 NRP-1在SCC9中呈高表达，A7R拮抗后SCC9细胞迁移能力明显下降，同时SCC9细胞上皮相关标记蛋白（epithelial cadherin，E-cad、β-catenin，β-cate）mRNA表达水平增高，而间充质相关标记蛋白（snail、fibronectin，FN、α-smooth muscle actin、α-SMA）mRNA表达水平下降。结论外源性拮抗SCC9细胞NRP-1可抑制其迁移能力并影响其细胞分化。

Abstract: Objective: To investigate the effect exogenous inhibition of Neuropilin-1(NRP-1) on the migration and the differentiation of the human tongue squamous carcinoma cell line SCC9. Methods: Western blot were performed to examine the expression of NRP-1 in cell line SCC9 and the human tongue squamous carcinoma cell cancer tissue. Besides, scratch test and transwell assay were employed to show the influence of exogenous inhibition of Neuropilin-1(NRP-1)on the migration of the SCC9 cells. And Real-time RT PCR were used to detect the expression of differentiation markers in the SCC9 cells exogenously inhibited of NRP-1. Results: NRP-1 express at high level in the SCC9 cells, and exogenous inhibition of NRP-1 depressed the migration of SCC9 cells. Real-time RT PCR showd the expression of epithelial cadherin and β-catenin increased, while the expression of snail, fibronectin and α-smooth muscle actin decreased. Conclusions: Exogenous inhibition of NRP-1 depressed the migration and differentiation of SCC9 cells.

中图分类号:

R739.9

查霖吴煜农郑灏赵宏惠安宁宋晓萌唐子春. 外源性拮抗神经纤毛蛋白1对人舌癌细胞SCC9细胞迁移及分化的影响[J]. , 2015, 6(4): 198-210.

参考文献

[1] Takagi S, Tsuji T, Amagai T, et al. Specific cell surface labels in the visual centers of Xenopus laevis tadpole identified using monoclonal antibodies [J]. Developmental Biology, 1987, 122(1):90–100.
[2] Luo Y,Raible D,Raper JA, et al.Collapsin:a protein in brain that induces the collapse and paralysis of neuronal growth cones[J].Cell,1993.75(2):p.217-27.
[3] He Z,Tessier-Lavigne M.Neuropilin is a receptor for the axonal chemorepellent Semaphorin III[J].Cell,1997.90(4):p.739-51.
[4] Wang L,Zeng H,Wang P,et al.Neuropilin-1-mediated vascular permeability factor/vascular endothelial growth factor-dependent endothelial cell migration[J].J Biol Chem,2003.278(49):p.48848-60.
[5] Whitaker GB,Limberg BJ,Rosenbaum JS.Vascular endothelial growth factor receptor-2 and neuropilin-1 form a receptor complex that is responsible for the differential signaling potency of VEGF(165) and VEGF(121) [J].J Biol Chem,2001.276(27):p.25520-31.
[6] Neufeld G, Cohen T, Gengrinovitch S, et al. Vascular endothelial growth factor (VEGF) and its receptors.[J]. Faseb Journal, 1999, 13:9-22.
[7] Vanveldhuizen PJ, Zulfigar M, Banerjee S, et al. Differential expression of neuropilin-1 in malignant and benign prostatic stromal tissue[J]. Oncol Rep, 2003. 10(5): p. 1067-71.
[8] Bielenberg DR, Pettaway CA, Takashima S, et al. Neuropilins in neoplasms: expression, regulation, and function. Exp Cell Res[J]. 2006. 312(5): p. 584-93.
[9] Ellis LM. The role of neuropilins in cancer[J]. Mol Cancer Ther, 2006. 5(5): p. 1099-107.
[10] Stephenson JM, Banerjee S, Saxena NK, et al. Neuropilin-1 is differentially expressed in myoepithelial cells and vascular smooth muscle cells in preneoplastic and neoplastic human breast: a possible marker for the progression of breast cancer[J]. Int J Cancer. 2002; 101(5):409-14.
[11] Staton C, Kumar I, Reed M, et al. Neuropilins in physiological and pathological angiogenesis[J]. J Pathol. 2007; 212(3): 237-48.
[12] Ochiumi T, Kitadai Y, Tanaka S, et al. Neuropilin-1 is involved in regulation of apoptosis and migration of human colon cancer[J]. Int J Oncol. 2006; 29(1): 105-16.
[13] Starzec A, Ladam P, Vassy R, et al. Structure-function analysis of the antiangiogenic ATWLPPR peptide inhibiting VEGF(165) binding to neuropilin-1 and molecular dynamics simulations of the ATWLPPR/neuropilin-1 complex[J]. Peptides, 2007. 28(12): p. 2397-402.
[14] Duband JL, Monier F, Delannet M, et al.Newgreen D. Epithelium-mesenchyme transition during neural crest development[J]. Acta Anatomica. 1995;154(1):63–78.
[15] Bolender DL, Markwald RR. Epithelial-mesenchymal transformation in chick atrioventricular cushion morphogenesis[J]. Scan Electron Microsc, 1979, 3:313-321.
[16] Griffith CM, Hay ED. Epithelial-mesenchymal transformation during palatal fusion: carboxyfluorescein traces cells at light and electron microscopic levels[J]. Development.1992;116(4):1087–1099.
[17] Yilmaz M, Christofori G. EMT, the cytoskeleton, and cancer cell invasion[J]. Cancer and Metastasis Reviews. 2009;28(1-2):15–33.
[18] Cao Y, Wang L, Nandy D, et al. Neuropilin-1 upholds dedifferentiation and propagation phenotypes of renal cell carcinoma cells by activating Akt and sonic hedgehog axes[J]. Cancer Res, 2008. 68(21): p. 8667-72.
[19] 张阳，傅振，宋晓萌，等. Semaphorin3A及其受体在舌鳞癌中的表达及作用[J]. 口腔生物医学，2011， 02（1）：21—25
[20] Wang ZL, Tang ZC, Zhang Y, et al. Neuropilin-1 down-regulation impairs cell migration and induces the differentiation of human tongue squamous cell carcinoma cells.[J]. Head Neck Oncol, 2012, 4: 54.
[21] Zhang Y, He X, Liu Y, et al. microRNA-320a inhibits tumor invasion by targeting neuropilin 1 and is associated with liver metastasis in colorectal cancer[J]. Oncol Rep. 2012; 27(3): 685-94.
[22] Mak P, Leav I, Pursell B, et al. ERbeta impedes prostate cancer EMT by destabilizing HIF-1alpha and inhibiting VEGF-mediated snail nuclear localization: implications for Gleason grading.[J]. Cancer Cell, 2010, 17(4):319-332.