CAJ | 학술논문

目的探讨强直性脊柱炎患者和健康对照组外周血单个核细胞已知微小RNA(miRNA)差异表达.方法分别构建10例强直性脊柱炎患者和9例健康对照组外周血单个核细胞小RNA文库,并运用新一代高通量Solexa测序技术,进行外周血单个核细胞已知miRNA的检测；运用实时荧光定量聚合酶链反应(FQ-PCR)进一步验证部分差异miRNA表达.结果在强直性脊柱炎患者与对照组中小RNA文库中,共获得小RNA总量分别是7511 859和10 178 958,比对上基因组部分小RNA分别是6052911 (80.58％)和8 476 243(83.27％)；已知miRNA分别是267和231个,通过miRNA差异性分析,129个上调miRNA和28个下调miRNA,其表达水平差异有统计学意义(P＜0.05)；FQ-PCR进一步验证了let-7b-3p、miR-146a-5p、miR-155-5p、let-7g-5p和miR-323a-5p差异表达水平与Solexa测序结果相似趋势.结论强直性脊柱炎患者外周血单个核细胞miRNA的差异表达,可能与强直性脊柱炎发病密切相关.
목적 탐토강직성척주염환자화건강대조조외주혈단개핵세포이지미소RNA(miRNA)차이표체.방법 분별구건10례강직성척주염환자화9례건강대조조외주혈단개핵세포소RNA문고,병운용신일대고통량Solexa측서기술,진행외주혈단개핵세포이지miRNA적검측；운용실시형광정량취합매련반응(FQ-PCR)진일보험증부분차이miRNA표체.결과 재강직성척주염환자여대조조중소RNA문고중,공획득소RNA총량분별시7511 859화10 178 958,비대상기인조부분소RNA분별시6052911 (80.58％)화8 476 243(83.27％)；이지miRNA분별시267화231개,통과miRNA차이성분석,129개상조miRNA화28개하조miRNA,기표체수평차이유통계학의의(P＜0.05)；FQ-PCR진일보험증료let-7b-3p、miR-146a-5p、miR-155-5p、let-7g-5p화miR-323a-5p차이표체수평여Solexa측서결과상사추세.결론 강직성척주염환자외주혈단개핵세포miRNA적차이표체,가능여강직성척주염발병밀절상관.
Objective To explore the differential expression of known microRNA (miRNA) expression in peripheral blood mononuclear cells (PBMCs) from patients with ankylosing spondylitis (AS)and healthy controls.Methods The microRNA library of PBMCs in 10 cases of AS and 10 cases of healthy controls were constructed,respectively.The samples of two groups were sequenced simultaneously using next generation high-throughput sequencing technology,and known miRNAs were analyzed differentially.Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to verify certain differential expression of known miRNA.Results A total of 7 511 859 and 10 178 958 small RNA reads were obtained in PBMCs from patients with AS and health controls 6 052 911 (80.58％) and 8 476 243 (83.27％) genome-matched reads,267 and 231 known miRNAs were identified in the library of AS and healthy controls,respectively.There were 157 known miRNAs with differential expression levels between the two libraries.FQ-PCR validated the similar trends with Solexa sequencing (let-7b-3p,miR-146a-5p,miR-155-5p,let-7g-5p and miR-323a-5p).Conclusion The differential expression levels of known miRNAs may be involved in the pathological process of AS.