山东大学耳鼻喉眼学报 ›› 2019, Vol. 33 ›› Issue (4): 66-70.doi: 10.6040/j.issn.1673-3770.0.2018.409

• 论著 • 上一篇    下一篇

LncRNA CTB-147C22.8对复发性呼吸道乳头状瘤细胞侵袭的影响

潘晓菲,王军,肖洋,马丽晶   

  1. 首都医科大学附属北京同仁医院耳鼻咽喉头颈外科, 耳鼻咽喉头颈科学教育部重点实验室(首都医科大学), 北京 100730
  • 出版日期:2019-07-20 发布日期:2019-07-22
  • 基金资助:
    国家重点研发计划项目(2017YFC0112500);北京市自然科学基金项目(7182036);儿科专项创新项目推广项目(XTCX201823)

Effects of lncRNA CTB-147C22.8 and mRNA KLK6 on the invasion of recurrent respiratory papilloma cells

PAN Xiaofei, WANG Jun, XIAO Yang, MA Lijing   

  1. Department of Otolaryngology, Head and Neck Surgery, Beijing Tongren Hospital affiliated to Capital Medical University, Key Laboratory of Otolaryngology, Head and Neck Science, Ministry of Education, Capital Medical University, Beijing 100730, China
  • Online:2019-07-20 Published:2019-07-22

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摘要： 目的 探讨lncRNA CTB-147C22.8对复发性呼吸道乳头状瘤细胞侵袭的影响。 方法 在前期高通量基因组表达谱芯片分析的基础上,建立人鼻咽上皮细胞系(NP69-SV40T),细胞培养后用脂质体LipofectaminTM2000转染,将含有外源基因的重组质粒pcDNA3.1/zeo+-CTB-147C22.8 转染人鼻咽上皮细胞内,根据转染载体,将细胞分为NP69-空白组、NP69-H145组、NP69-H6373组3组,NP69-空白组指未转染质粒的细胞,NP69-H145组指转染空白质粒的细胞,NP69-H6373组指转染了目的基因CTB-147C22.8的细胞。采用qRT-PCR技术检测转染lncRNA CTB-147C22.8前后人鼻咽上皮细胞中KLK6表达;对转染了lncRNA CTB-147C22.8的上皮细胞进行流式细胞检测,用FlowJo software进行细胞周期分析;采用Transwell实验检测lncRNA CTB-147C22.8对细胞侵袭的影响。 结果 KLK6基因相对表达量NP69-空白组、NP69-H145组、NP69-H6373组分别为1.005 771 107±0.008 724 979、1.591 421 113±0.311 999 127、1.880 766 963±0.109 759 031,NP69-H6373组KLK6基因相对表达量与NP69-空白组、NP69-H145组比较差异有统计学意义(F=10.892, P=0.01)。NP69-H6373组S期所占比例与NP69-空白组、NP69-H145组比较S期延长,差异有统计学意义(F=8.763, P=0.017)。NP69-空白组、NP69-H145组和NP69-H6373组9个视野下观察到的细胞迁移数分别为(6.889±2.261)、(8.222±3.114)、(39.556±18.228)个,NP69-H6373组细胞迁移数与NP69-空白组、NP69-H145组比较差异有统计学意义(F=3.038, P=0.034)。 结论 lncRNA CTB-147C22.8能促进复发性呼吸道乳头状瘤细胞侵袭。

关键词: 复发性呼吸道乳头状瘤, 长链非编码 RNA, KLK6基因, 细胞侵袭

Abstract: Objective To investigate the effect of lncRNA CTB-147C22.8 on the invasion of recurrent respiratory papilloma cells. Methods A high-throughput genome expression profile chip was used for early stage analysis. To establish a human nasopharyngeal epithelial cell line(NP69 - SV40T), cells were incubated with Lipofectamine TM2000 for transfection; the transfected nasopharyngeal epithelial cells contained the exogenous gene recombinant plasmid pcDNA3.1/zeo+-CTB-147C22.8. Based on the transfection carrier, cells were divided into three groups: NP69 blank group, NP69 H145 group, and NP69-H6373 group. NP69 indicates untransfected plasmid cell blank group, NP69 H145 group refers to the cells transfected with a blank plasmid, and NP69 H6373 group refers to the cells transfected with the target gene CTB-147C22.8. KLK6 expression in human nasopharyngeal epithelial cells before and after transfection of lncRNA CTB-147C22.8 was detected by qrt-pcr. Flow cytometry was performed on the epithelial cells transfected with lncRNA CTB-147C22.8, and cell cycle analysis was performed by using FlowJo software. Transwell assay was used to determine the effect of lncRNA CTB-147C22.8 on cell invasion. Results KLK6 gene expression levels in the NP69-blank, NP69-H145, and NP69 H6373 groups were 1.005 771 107±0.008 724 979, 1.591 421 113±0.311 999 127, and 1.880 766 963±0.109 759 031, respectively. Compared with NP69-blank and NP69-H145 groups, the duration of cells in S stage in the NP69 H6373 group was significantly longer(F=8.763, P=0.017). The number of cells migrating in the NP69 H6373, NP69-H145, and NP69 H6373 groups was 6.889±2.261, 8.222±3.114, and 39.556±18.228, respectively. The differences in these numbers were statistically significant between the NP69 H6373 group, and the NP69-blank group and NP69-H145 group(F=3.038, P=0.034). Conclusion lncRNA CTB-147C22.8 promotes the invasion of recurrent respiratory papilloma cells.

Key words: Recurrent respiratory papillomatosis, lncRNA, KLK6, Invasion

中图分类号: 

  • R734
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