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
潘晓菲,王军,肖洋,马丽晶
PAN Xiaofei, WANG Jun, XIAO Yang, MA Lijing
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。
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