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目的研究先天性巨结肠(HSCR)患儿疾病相关的长链非编码RNA(lnc RNA)的表达情况,为进一步研究其参与的发病机制打下基础。方法收集2015-01-01—2016-01-01于第四军医大学唐都医院就诊,经手术和病理确诊为HSCR的无神经节细胞肠道全层组织与正常对照肠组织各30例(3例用于基因芯片筛选,27例用于扩大样本验证)。通过基因芯片技术筛选获得HSCR相关的lnc RNA表达谱。进一步筛选通过提高入选条件(探针荧光初始信号值≥100,差异倍数≥3),并采用实时荧光定量(q RT-PCR)验证筛选出的lnc RNA在27例样本中的表达情况,将差异有显著统计学意义的基因确定为与疾病明确相关的生物标记性lnc RNA。结果通过基因芯片技术筛选初步获得217个HSCR相关的差异表达lnc RNA(差异倍数≥2,P<0.05),其中上调13个,下调204个。进一步筛选后总共获得2个lnc RNA,其表达量的差异经扩大样本验证后在病变组织与对照组织中有统计学意义(P<0.05)。结论 lnc RNA差异表达谱与HSCR的发病密切相关,为疾病的基因调控研究提供了新的方向。
Objective To study the expression of disease-associated long-chain non-coding RNA (lnc RNA) in children with Hirschsprung’s disease and to lay a foundation for further study of its pathogenesis. Methods Collecting 2015-01-01-2016-01-01 At the Tangdu Hospital of the Fourth Military Medical University, 30 cases of ganglion cell full-thickness tissues and normal control intestinal tissues were identified by surgery and pathology respectively Example for gene chip screening, 27 cases used to expand the sample validation). HSCR-related lnc RNA expression profiles were obtained by gene chip technology. Further screening confirmed that the lnc RNA was screened out in 27 samples by increasing the selection conditions (probe initial fluorescence signal≥100 and the multiple of multiple≥3), and using real-time fluorescence quantitative PCR (q RT-PCR) Significantly statistically significant genes were identified as biomarker lnc RNAs that were clearly associated with the disease. Results A total of 217 HSCR-related differentially expressed lnc RNAs were screened by gene chip technique (with a multiple of ≥2, P <0.05), of which 13 were up-regulated and 204 were down-regulated. After further screening, a total of 2 lnc RNAs were obtained, and the differences of their expression levels were statistically significant (P <0.05) in the diseased and control tissues after the expanded samples were verified. Conclusion The differential expression profile of lnc RNA is closely related to the pathogenesis of HSCR, providing a new direction for gene regulation of disease.