Effects of silencing <i>Snail1</i> gene on the expression of tight junction proteins and the migration ability of Hep-2 cells _Journal of Biomedical Engineering

Authors：

LIU Shuangfeng ^1,2 , SHEN Yang ² , FENG Tang ² ,  LIU Xiaoheng ²

1. School of Medical Laboratory Science, Chengdu Medical College, Chengdu 610500, P.R.China;
2. Institute of Biomedical Engineering, School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, P.R.China;

Corresponding author：

LIU Xiaoheng, Email: liuxiaohg@scu.edu.cn

Keywords：

Snail1; gene silence; Hep-2 cell; tight junction; cell migration

DOI：

10.7507/1001-5515.201702005

Video：

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To investigate the effects of Snail1 gene silence on the expression of tight junction proteins and the migration ability of Hep-2 cells, Hep-2 cells were transfected with plasmids which is containing the shRNA of Snail1 gene, and cultured till the cells could be passaged stably (named Sh-snail1 cells). The expression of tight junction proteins (ZO-1, Occludin, Claudin-5) were detected by Western blot. The migration ability of Sh-snail1 cells was investigated by wound healing assay, and the protein expression of members of RhoGTPase family (RhoA, Cdc42) was detected by Western blot, which is closely related to the migration ability. Our results showed that the expression of tight junction proteins (ZO-1, Occludin, Claudin-5) was significantly increased; the migration ability of Sh-snail1 cell was inhibited; the expression of RhoA and Cdc42 was downregulated. All of these indicated that silencing the gene of Snail1 in Hep-2 cells can up-regulate the expression of tight junction proteins and down regulate the expression of Cdc42 and RhoA, and further inhibit the migration of Hep-2 cells. Furthermore, opening of the tight junctions between cells and the stronger migration ability of cancer cells are important processes in cancer metastasis. It is confirmed that the Snail1 gene is closely related to the two processes, providing an experimental basis for targeted therapy of laryngeal squamous cell carcinoma.

Citation： LIU Shuangfeng, SHEN Yang, FENG Tang, LIU Xiaoheng. Effects of silencing Snail1 gene on the expression of tight junction proteins and the migration ability of Hep-2 cells . Journal of Biomedical Engineering, 2017, 34(4): 591-596. doi: 10.7507/1001-5515.201702005 Copy

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11.	Yilmaz M, Christofori G. EMT, the cytoskeleton and cancer cell invasion. Cancer Metastasis Rev, 2009, 28(1/2): 15-33.
12.	Yeo D, He Hong, Baldwin G S, et al. The role of p21-activated kinases in pancreatic cancer. Pancreas, 2015, 44(3): 363-369.
13.	Shirley S H, Hammiller B, Hansen L A, et al. Role of the epidermal growth factor receptor in ultraviolet radiation induction of Snail family transcription factors. J Dermatol Sci, 2014, 76(2): 149-151.
14.	Zhao Xinyu, Li Lei, Wang Xiaobo, et al. Inhibition of snail family transcriptional repressor 2 (SNAI2) enhances multidrug resistance of hepatocellular carcinoma cells. PLoS One, 2016, 11(10): e0164752.

1. Markou K, Christoforidou A, Karasmanis I, et al. Laryngeal cancer: epidemiological data from Northern Greece and review of the literature. Hippokratia, 2013, 17(4): 313-318.
2. Iovănescu G H, Poenaru M, Doroş C, et al. Histopathological prognostic and risk factors in patients with laryngeal neoplasms. Rom J Morphol Embryol, 2013, 54(4): 1087-1092.
3. 郭世伟, 李先鹏, 吴义峰, 等. E-cadherin 在胰腺癌中的表达及其对预后的影响. 现代生物医学进展, 2013, 13(1): 102-106.
4. Liu Shuangfeng, Zhou Fating, Shen Yang, et al. Fluid shear stress induces epithelial-mesenchymal transition (EMT) in Hep-2 cells. Oncotarget, 2016, 7(22): 32876-32892.
5. Fan Fan, Samuel S, Evans K W, et al. Overexpression of snail induces epithelial-mesenchymal transition and a cancer stem cell-like phenotype in human colorectal cancer cells. Cancer Med, 2012, 1(1): 5-16.
6. Lamouille S, Connolly E, Smyth J W, et al. TGF-β-induced activation of mTOR complex 2 drives epithelial-mesenchymal transition and cell invasion. J Cell Sci, 2012, 125(Pt 5): 1259-1273.
7. Wen Sijian, Zhang Wei, Ni Nana, et al. Expression of Rho GTPases family in melanoma cells and its influence on cytoskeleton and migration. Oncotarget, 2017, 8(18): 30112-30122.
8. Amin E, Dubey B N, ZHANG Sicai, et al. Rho-kinase: regulation, (dys) function, and inhibition. Biol Chem, 2013, 394(11): 1399-1410.
9. Guo Huimin, Zhang Xiaoqi, Xu Chunhong, et al. Inhibition of invasion and metastasis of gastric cancer cells through snail targeting artificial microRNA interference. Asian Pac J Cancer Prev, 2011, 12(12): 3433-3438.
10. Huang R Y, Guilford P, Thiery J P. Early events in cell adhesion and polarity during epithelial-mesenchymal transition. J Cell Sci, 2012, 125(19): 4417-4422.
11. Yilmaz M, Christofori G. EMT, the cytoskeleton and cancer cell invasion. Cancer Metastasis Rev, 2009, 28(1/2): 15-33.
12. Yeo D, He Hong, Baldwin G S, et al. The role of p21-activated kinases in pancreatic cancer. Pancreas, 2015, 44(3): 363-369.
13. Shirley S H, Hammiller B, Hansen L A, et al. Role of the epidermal growth factor receptor in ultraviolet radiation induction of Snail family transcription factors. J Dermatol Sci, 2014, 76(2): 149-151.
14. Zhao Xinyu, Li Lei, Wang Xiaobo, et al. Inhibition of snail family transcriptional repressor 2 (SNAI2) enhances multidrug resistance of hepatocellular carcinoma cells. PLoS One, 2016, 11(10): e0164752.

Journal of Biomedical Engineering

Effects of silencing Snail1 gene on the expression of tight junction proteins and the migration ability of Hep-2 cells

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