CAJ | 학술논문

背景与目的：微小RNA(microRNA，miRNA)是一类存在于真核生物体内只有19~39 bp大小的内源性非编码RNA，它能在转录和翻译水平调控基因的表达，在细胞的增殖分化、新陈代谢、免疫调控和凋亡等方面起着重要的作用。本研究检测miR-216a-5p在肺癌组织和肺癌细胞系的表达并探讨其对肺癌细胞侵袭能力的影响及其调控机制。方法：使用实时荧光定量PCR(quantitative real-time PCR，qRT-PCR)检测55例肺癌患者的肺癌组织和7种肺癌细胞系中miR-216a-5p的表达情况；miR-216a-5p瞬时转染A549、95D和H4603种肺癌细胞系，使用Transwell侵袭实验检测miR-216a-5p对肺癌细胞系侵袭能力的影响；预测并构建miR-216a-5p的候选靶基因基质金属蛋白酶16(matrix metalloproteinase 16，MMP16)基因的双荧光素酶报告基因表达质粒，使用qRT-PCR和蛋白[质]印迹法(Western blot)检测miR-216a-5p对靶基因MMP16的mRNA和蛋白表达的影响；小干扰RNA(siRNA)干扰MMP16与上调miR-216a-5p对比检测其对肺癌细胞侵袭能力的影响。结果：90.91%(50/55)的肺癌患者肿瘤组织中miR-216a-5p表达明显低于对应的癌旁组织(P<0.05)。7种肺癌细胞系中miR-216a-5p的表达量仅为对照组的7.00%~32.00%(P<0.05)。上调miR-216a-5p的表达能够抑制肺癌细胞的侵袭；siRNA干扰MMP16与转染上调miR-216a-5p都能够抑制肺癌细胞中MMP16的表达，并抑制肺癌细胞侵袭。结论：miR-216a-5p可以作为临床肺癌诊断的候选标志物之一，并且其能够通过下调MMP16的表达从而抑制肺癌细胞的侵袭。
배경여목적：미소RNA(microRNA，miRNA)시일류존재우진핵생물체내지유19~39 bp대소적내원성비편마RNA，타능재전록화번역수평조공기인적표체，재세포적증식분화、신진대사、면역조공화조망등방면기착중요적작용。본연구검측miR-216a-5p재폐암조직화폐암세포계적표체병탐토기대폐암세포침습능력적영향급기조공궤제。방법：사용실시형광정량PCR(quantitative real-time PCR，qRT-PCR)검측55례폐암환자적폐암조직화7충폐암세포계중miR-216a-5p적표체정황；miR-216a-5p순시전염A549、95D화H4603충폐암세포계，사용Transwell침습실험검측miR-216a-5p대폐암세포계침습능력적영향；예측병구건miR-216a-5p적후선파기인기질금속단백매16(matrix metalloproteinase 16，MMP16)기인적쌍형광소매보고기인표체질립，사용qRT-PCR화단백[질]인적법(Western blot)검측miR-216a-5p대파기인MMP16적mRNA화단백표체적영향；소간우RNA(siRNA)간우MMP16여상조miR-216a-5p대비검측기대폐암세포침습능력적영향。결과：90.91%(50/55)적폐암환자종류조직중miR-216a-5p표체명현저우대응적암방조직(P<0.05)。7충폐암세포계중miR-216a-5p적표체량부위대조조적7.00%~32.00%(P<0.05)。상조miR-216a-5p적표체능구억제폐암세포적침습；siRNA간우MMP16여전염상조miR-216a-5p도능구억제폐암세포중MMP16적표체，병억제폐암세포침습。결론：miR-216a-5p가이작위림상폐암진단적후선표지물지일，병차기능구통과하조MMP16적표체종이억제폐암세포적침습。
Background and purpose:MicroRNA (miRNA) belongs to a class of 19 to 30 nucleotide-long, endogenous noncoding RNA expressed in eukaryotes and predominantly inhibits gene expression at the post-transcriptional level. The miRNAs play critical roles in cell proliferation and differentiation, apoptosis, metabolism, and immune regulation. This study aimed to detect the expression of miR-216a-5p in lung cancer tissues and lung cancer cell lines, and to discuss the effects of miR-216a-5p on the invasion ability of lung cancer cells and the mechanism.Methods:Quantitative real-time PCR (qRT-PCR) was used to detect the expression of miR-216a-5p in lung cancer tissues of 55 cases and 7 lung cancer cell lines. Three lung cancer cell lines of A549, 95D and H460 were transiently transfected by miR-216a-5p, and Transwell was used to detect the effects of miR-216a-5p on the invasion of lung cancer cell lines. The dual luciferase reporter plasmids containing the miR-216a-5p candidate target gene and the gene of matrix metalloproteinase 16 (MMP16) were predicted and constructed. qRT-PCR and Western blot were used to detect the changes in mRNA and protein levels of target geneMMP16 by miR-216a-5p. The interference of MMP16 by siRNA and up-regulation miR-216a-5p by transfection were compared on the invasion of lung cancer cells.Results:The miR-216a-5p expression levels were all signiifcantly reduced in 90.91% (50 of 55 patients) tumor tissues compared with corresponding adjacent normal lung tissues (P<0.05). The miR-216a-5p expression levels were only 7.00%-32.00%in 7 lung cancer cells compared with the control group (P<0.05). Up-regulation of the expression of miR-216a-5p inhibited the invasion of lung cancer cells; interference of MMP16 by siRNA, as well as up-regulating miR-216a-5p by transfection, inhibited the expression of MMP16 in lung cancer leading to inhibition of the invasion of lung cancer cells. Conclusion:miR-216a-5p can be a candidate marker in clinical diagnosis and it can inhibit the invasion of lung cancer cells by down-regulating the expression of MMP16.