Testin基因对高转移鼻咽鳞状细胞癌的影响

钟准; 尹述成

doi:10.13201/j.issn.1001-1781.2017.21.013

Testin基因对高转移鼻咽鳞状细胞癌的影响

钟准^1,2,
尹述成^1, ,

- 武汉大学中南医院耳鼻咽喉头颈外科(武汉, 430071)
基金项目:
湖北省自然科学基金资助项目 (No:2009CBD420)

详细信息

通讯作者: 尹述成, E-mail:2014105529@qq.com

中图分类号: R739.6

收稿日期: 2017-06-29

Role of Testin in nasopharyngeal squamous cancer with high ability of metastasis

ZHONG Zhun^1,2,
YIN Shucheng^1, ,

- Department of Otolaryngology Head and Neck Surgery, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China

More Information

Corresponding author: YIN Shucheng, E-mail: 2014105529@qq.com

Received Date: 29 June 2017

摘要

摘要: 目的: 探讨Testin(TES)基因对人鼻咽鳞状细胞癌株5-8F细胞增殖、迁移的影响。方法: 以RT-PCR法从人鼻咽鳞状细胞癌株5-8F细胞获得目的基因,构建真核表达载体pEGFP-N1-TES,采用PCR、基因测序等对其进行鉴定;然后将其转导入人高转移鼻咽鳞状细胞癌株5-8F中,通过RT-PCR法及Western-blot法对其进行鉴定。接着采用流式细胞术、划痕试验检测转染后5-8F细胞增殖及迁移能力的影响。结果: 转染外源性TES基因后,转染外源性TES基因的5-8F细胞呈现明显的凋亡,差异有统计学意义(P<0.05)。细胞划痕实验表明:在第12、24、48 h时,TES组细胞迁移速率明显比空载体组和未转染组要慢(P<0.01)。结论: 该研究成功建立了能稳定高表达TES的细胞模型。TES基因在体外可明显抑制鼻咽癌5-8F细胞的增殖和降低其迁移运动能力,可能是一种潜在的抑癌基因。
- Testin基因 /
- 鳞状细胞癌 /
- 基因转染
Abstract: Objective: To explore the influence and regulatory mechanism of TES gene on proliferation and migration of nasopharyngeal squamous cancer(NSPC) 5-8F cell.Method: DNA fragment encoding TES was obtained by RT-PCR method from the human highly metastatic nasopharyngeal squamous carcinoma cell line 5-8F. we identified the recombinant plasmid pEGFP-N1-TES by RT-PCR and DAN sequencing. we stablely transfected the pEGFP-N1-TES into the human highly metastatic nasopharyngeal squamous carcinoma cell line 5-8F, and detected the expression of TES by the RT-PCR and Western-blot method. And detected the impact of 5-8F cells transfection by flow cytometry and scratch tests.Result: Flow cytometry analysis showed that the apoptotic in 5-8F/pEGFP- N1-TES was significantly higher than non-transected TES and 5-8F/pEGFP-N1, and the differences were statistically significant(P<0.05). Cell scratch experiments showed that the 5-8F/pEGFP-N1-TES group cell migration rate was obviously lower than non-transected TES and 5-8F/pEGFP-N1 group in the first 12 h, 24 h and 48 h.The difference was significant(P<0.01).Conclusion: The stable transfectant cell model was established successfully. TES in vitro could significantly increase apoptosis and reduce the athletic ability. And thus TES gene might be a novel candidate of tumor-suppressor.
- Testin gene /
- nasopharyngeal squamous cancer /
- gene transfection

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参考文献(15)

[1]	SARTI M, PINTON S, LIMONI C, et al.Differential expression of Testinand Survivin in breast cancer subtypes[J].Oncol Rep, 2013, 30:824-832.
[2]	SARTI M, SEVIGNANI C, CALIN G A, et al.Adenovirla transduction of Testin gene into breast and uterine cancer cell lines promotes apoptosis and tumor reduction in vivo[J].Clin Cancer Res, 2005, 11:806-813.
[3]	ARCHACKI S R, ANGHELOIU G, MORAVEC CS, et al.Comparative gene expression analysis between coronary arteries and internal mammary arteries identifies a role for the TES gene in endothelial cell functions relevant to coronary artery disease[J].Med JExp Clin Res, 2012, 21:1364-1373.
[4]	QIU H, ZHU J, YUAN C, et al.Frequent hypermethylation and loss of heterozygosity of the testis derived transcript gene in ovarian cancer[J].Cancer Science, 2010, 101:1255-1260.
[5]	GU Z, DING K, LIANG K, et al.TESTIN suppresses tumor growth and invasion via manipulating cell cycle progression in endometrial carcinoma[J].Med Sci Monit, 2014, 20:980-987.
[6]	WANG M, WANG Q, PENG W J, et al.Testin is a tumor suppressor in non-small cell lung cancer[J].Oncol Rep, 2017, 37:1027-1035.
[7]	LATIL A, CUSSENOL O, FOURNIER G, et al.Loss of heterozygosity at 7q3l is a frequent and early event in prostate cancer[J].Clin Cancer Res, 1995, 1:1385-1389.
[8]	MATSUURA K, SHIGA K, YOKOYAMA J, et al.Loss of heterozygosity of chromosome 9p21 and 7q3l is correlated with high incidence of recurrent tumor in head and neck squamous cell carcinoma[J].Anticancer Res, 1998, 18:453-458.
[9]	张飞, 尹述成.Testin基因在鼻咽癌组织中的表达及其临床意义[J].临床耳鼻咽喉头颈外科杂志, 2016, 30 (12):982-985.
[10]	BINGLE C D, CRAVEN C J.Meet the relatives:a family of BPI-and LBP-related proteins[J].Trends Immunol, 2004, 25:53-55.
[11]	ZHOU H D, LI G Y, YANG Y X, et al.Intracellular colocalization of SPLUNCI protein with nanobacteria in nasopharyngeal carcinoma epithelia HNE1 cells depended on the bactericidal permeability increasing protein domain[J].Mol Immunol, 2006, 43:1864-1871.
[12]	陈斌, 殷善开.鼻咽癌的病因及流行病学状况[J].全科医疗专题讨研, 2002, 5 (4):262-263.
[13]	TOBIAS E S, HURLSTONE A F, MACKENZIE E, et al.The TES gene at 7q31.1 is methylated in tumours and encodes a novel growth-suppressing LIMdomain protein[J].Oncogene, 2001, 20:2844-2853.
[14]	MANUELA S, SANDRA P, COSTANZO L, et al.Differential expression of testin and survin in breast cancer subtypes[J].Oncol Rep, 2013, 30:824-832.
[15]	DRUSCO A, ZANESI N, ROLDO C, et al.Knockout mice reveal a tumor suppressor function for Testin[J].PNAS, 2005, 102:10947-10951.