干扰内皮素-1可抑制鼻咽癌细胞增殖、迁移和侵袭

发布时间：2017-12-27 20:22

本文关键词：干扰内皮素-1可抑制鼻咽癌细胞增殖、迁移和侵袭　出处：《南方医科大学学报》2016年07期 　论文类型：期刊论文

【摘要】：目的探讨干扰内皮素-1(ET-1)基因对鼻咽癌细胞增殖、迁移及侵袭的影响以及相应的分子机制。方法构建ET-1sh RNA慢病毒干扰载体,成功包装慢病毒后感染鼻咽癌5-8F细胞。Western blotting检测ET-1干扰效率。利用MTT、细胞周期检测、平板克隆形成实验、Transwell小室、Boyden小室以及裸鼠皮下成瘤实验分别检测细胞增殖、迁移、侵袭能力的改变。Western blotting检测上皮间质转化(EMT)相应基因表达改变。结果与对照组细胞相比,鼻咽癌细胞5-8F感染慢病毒sh RNAET-1后ET-1的表达水平显著降低。MTT实验结果显示,在抑制ET-1表达后细胞增殖能力显著下降。流式细胞仪检测细胞周期显示干扰ET-1表达,G1细胞数明显增多。平板克隆形成实验结果显示干扰ET-1表达,癌细胞克隆形成能力显著下降。皮下裸鼠成瘤实验结果同样显示,ET-1表达抑制后癌细胞成瘤能力显著下降。Transwell和Boyden小室结果表明,ET-1表达沉默后可显著降低细胞的迁移和侵袭能力。通过检测ET-1沉默后EMT相关分子的表达改变,结果显示上皮标志物E-cadherin和CK18表达增高,而间质标志物Vimentin和N-cadherin表达降低。结论 ET-1在鼻咽癌中可通过调控上皮间质转化相关基因促进细胞增殖、迁移和侵袭能力。
[Abstract]:Objective to investigate the effects of endothelin -1 (ET-1) gene on proliferation, migration and invasion of nasopharyngeal carcinoma cells and its molecular mechanism. Methods the ET-1sh RNA lentivirus interference carrier was constructed and the 5-8F cells of nasopharyngeal carcinoma were infected after the lentivirus was successfully packaged. Western blotting detection of ET-1 interference efficiency. The proliferation, migration and invasion ability of cells were detected by MTT, cell cycle detection, plate clone formation test, Transwell chamber, Boyden chamber and subcutaneous tumor formation in nude mice. Western blotting was used to detect the changes in the expression of corresponding gene expression in epithelial mesenchymal transition (EMT). Results compared with the control group, the expression level of ET-1 was significantly decreased after 5-8F infection of lentivirus sh RNAET-1 in nasopharyngeal carcinoma cells. The results of MTT experiment showed that the cell proliferation ability decreased significantly after inhibiting the expression of ET-1. The cell cycle of flow cytometry showed the expression of interference ET-1, and the number of G1 cells increased significantly. The experimental results of the formation of clones showed that the expression of interfering ET-1 and the ability to clone the cancer cells decreased significantly. The results of tumor formation in subcutaneous nude mice also showed that the tumor cell forming ability of cancer cells decreased significantly after the inhibition of ET-1 expression. The results of Transwell and Boyden compartments showed that the expression of ET-1 could significantly reduce the cell migration and invasion ability. By detecting the expression changes of EMT related molecules after ET-1 silencing, the results showed that the expression of E-cadherin and CK18 increased, while the expression of interstitial markers Vimentin and N-cadherin decreased. Conclusion ET-1 can promote cell proliferation, migration and invasion by regulating epithelial mesenchymal transition related genes in nasopharyngeal carcinoma.
【作者单位】：汕头大学医学院第一附属医院耳鼻咽喉科;南方医科大学南方医院医务处;南方医科大学南方医院耳鼻咽喉头颈外科;
【基金】：国家自然科学基金(81301849)~~
【分类号】：R739.63
【正文快照】： 鼻咽癌是我国南方及东南亚地区一种常见的恶性肿瘤,临床以低分化、高转移为主要特征,临床上75%~90%的鼻咽癌病人初诊时已是局部晚期并发生颈部淋巴结转移[1]。尽管以调强放疗技术为主的放化疗综合治疗使鼻咽癌的局控率有明显提高,但目前临床上仍无法对鼻咽癌转移进行早期预测

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