中国全科医学2016，Vol.19Issue(21)：2513-2517,5.DOI:10.3969/j.issn.1007-9572.2016.21.005
miR -150促进鼻咽癌细胞增殖和侵袭的机制研究
Mechanism Research of miR-150 Promoting the Proliferation and Invasion of Nasopharyngeal Carcinoma
摘要
Abstract
Objective To investigate whether miR - 150 promotes cell proliferation and invasion by target regulating of programmed cell death 4 ( PDCD4 ) or not, and further reveal the oncogeme function of miR - 150 in nasopharyngeal carcinoma. Methods From March to June 2014,we cultured the nasopharyngeal carcinoma CNE - 2,and selected the well -grown ones in the experiment. We designed and synthesized PDCD4 of wild type primer sequence and mutant primer sequence, which would be linked to the carrier vector with luciferase reporter gene,and luciferase activity was detected. Vector,pcDNA3. 1 ( + ) - PDCD4,miR - ctr,and miR - 150 mimics combined with pcDNA3. 1( + ) - PDCD4 were transient transfected into CNE - 2 respectively,and the expression level of PDCD4 was detected after 48 h by Western blotting method. Vector, pcDNA3. 1( + ) - PDCD4,miR - 150 inhibitors and miR - 150 mimics combined with pcDNA3. 1( + ) - PDCD4 were transient tranfected into CNE - 2 respectively,and respectively cultured for 24,48,72,and 96 h to detect the absorbance (OD)value to reflect its ascending ability. Vector,pcDNA3. 1( + ) - PDCD4,miR - 150 inhibitors and miR - 150 mimics combined with pcDNA3. 1( + ) - PDCD4 were transient transfected into CNE - 2 respectively,and the invasion ability of CNE- 2 was reflected by Transwell invasion experiment. Results The luciferase activity of PDCD4 of wild type cell lines that without miR - 150 transfection was(0. 975 ± 0. 112),and luciferase activity of PDCD4 of wild type cell lines that with miR - 150 transfection was(0. 588 ± 0. 042),which showed significant differences( t = 7. 853,P = 0. 018). The luciferase activity of PDCD4 of mutant type cell lines that without miR - 150 transfection was(0. 992 ± 0. 135),mutant type cell lines that with miR - 150 transfection was( 0. 875 ± 0. 095 ),which showed no significant differences( t = 1. 461,P = 0. 281 ) . The expression levels of PDCD4 that were transient transfected with Vector,pcDNA3. 1( + ) - PDCD4,miR - ctr,miR - 150 mimics combined with pcDNA3. 1( + ) - PDCD4 were(0. 665 ± 0. 058), (1. 147 ± 0. 152), (0. 074 ± 0. 068),and (0. 313 ± 0. 036)respectively,which showed significant differences(F = 43. 410,P ﹤ 0. 011);the expression level of CNE -2 PDCD4 that had been transfected with Vector,miR - ctr,miR - 150 mimics combined with pcDNA3. 1( + ) - PDCD4 was lower than that of the CNE - 2 with transfection of pcDNA3. 1( + ) - PCDC4,which showed significant differences( P ﹤0. 05). The transfected objects and time played an interactive role in the proliferation capacity of CNE - 2,and both of them had a main effect in the proliferation capacity of CNE - 2(P ﹤ 0. 001). The numbers of transmembrane cells of CNE - 2 that were transient transfected with Vector,pcDNA3. 1( + ) - PDCD4,miR - 150 inhibitors and miR - 150 mimics combined with pcDNA3. 1( + ) - PDCD4 were(56. 6 ± 7. 5), (26. 5 ± 3. 7), (30. 5 ± 4. 7),and(55. 2 ± 6. 9),which showed significant differences(F = 18. 550,P = 0. 014);the number of transmembrane cells of CNE - 2 that were transfected with pcDNA3. 1( + ) - PDCD4,and miR - 150 inhibitors was less than that of CNE - 2 that were transfected with Vector, miR - 150mimics combined with pcDNA3. 1( + ) - PDCD4,which showed significant differences(P ﹤ 0. 05). Conclusion miR - 150 enhances the proliferation and invasion capacity of nasopharyngeal carcinoma cells by inhibiting PDCD4 expression.关键词
鼻咽肿瘤/miR-150/程序性细胞死亡因子4/细胞增殖/侵袭Key words
Nasopharyngeal neoplasms/miR - 150/PDCD4/Cell proliferation/Invasion分类
医药卫生引用本文复制引用
张先锋,黄远见,肖娟,张志伟,黄卫国..miR -150促进鼻咽癌细胞增殖和侵袭的机制研究[J].中国全科医学,2016,19(21):2513-2517,5.基金项目
国家自然科学基金青年基金资助项目（81100568）；湖南省科技计划项目 （）
