激肽释放酶6在成人型复发性呼吸道乳头状瘤发病机制中作用的实验研究

Experimental study on the role of kallikrein 6 in the pathogenesis of adult-onset recurrent respiratory papillomatosis

目的探索激肽释放酶6(kallikrein6,KLK6)在成人型复发性呼吸道乳头状瘤病(adult-onset recurrent respiratory papillomatosis,AORRP)发病中的作用,明确其对上皮细胞增殖、侵袭的影响。方法在前期基因芯片分析结果提示喉乳头瘤组织KLK6高表达的基础上,构建KLK6基因干扰慢病毒载体感染鼻咽上皮细胞,CCK8法检测细胞增殖能力,RT-PCR检测基质金属蛋白酶12(matrix metalloproteinase 12,MMP12)和血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达,扫描电镜观察细胞形态。结果实验组细胞增殖能力较空白组、对照组减弱(F24h=983.543,F48h=3382.532,F72h=250.484,P均<0.05)。MMP12 mRNA在空白组表达量为0.984±0.038,对照组为3.816±0.091,均高于实验组0.649±0.036,差异有统计学意义(P<0.05);VEGF在实验组的表达量为0.527±0.052,较空白组(0.975±0.055)和对照组(1.211±0.130)明显降低,差异有统计学意义(P<0.05)。电镜下观察实验组细胞紧密连接数量较空白组增多。结论 KLK6可能通过同向调控MMP12、VEGF增强上皮细胞的增殖和侵袭能力,促进AORRP的发生。

OBJECTIVE To explore the role of kallikrein 6(KLK6) in the pathogenesis of adult-onset recurrent respiratory papillomatosis and its effect on the proliferation and invasion of epithelial cells.METHODSThe early gene chip analysis suggested high expression of KLK6 in AORRP.Lentiviral vector interfering with KLK6 gene was constructed to infect nasopharyngeal epithelial cells.The cell proliferating vitality were detected by cell counting kit 8,the expression of MMP12 and VEGF mRNA were detected by Real Time PCR,and cell morphology changes were observed by electronic microscope.RESULTSThe cell proliferation ability of the experimental group was significantly weaker than the blank group and the control group(F24h=983.543,F48h=3382.532,F72h=250.484,all P<0.05).The expression of MMP12 mRNA in the blank group was 0.984±0.038 and that in the control group was 3.816±0.091,which was significantly higher than that in the experimental group (0.649±0.036).The expression of VEGF in the experimental group was 0.527±0.052,which was significantly lower than that in the blank group(0.975±0.055) and the control group(1.211±0.130).Under electronic microscope,the number of tight junctions in the experimental group was more than that in the blank group.CONCLUSION KLK6 may enhance the abilities of proliferation and invasion of epithelial cells by regulating MMP12 and VEGF in the same direction,and promote the occurrence of adult-onset recurrent respiratory papillomatosis.