Screening miRNAs for early diagnosis of colorectal cancer by small RNA deep sequencing and evaluation in a Chinese patient population.
Screening miRNAs for early diagnosis of colorectal cancer by small RNA deep sequencing and evaluation in a Chinese patient population.
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通过小RNA深度测序筛选用于结直肠癌早期诊断的miRNA并在中国患者群体中进行评估
DOI：
10.2147/ott.s100427
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发表时间：
2016
影响因子：
4
通讯作者：
Tang G
中科院分区：
文献类型：
--
作者：
Wang X;Chen L;Jin H;Wang S;Zhang Y;Tang X;Tang G
目的本研究旨在通过对中国结直肠癌患者的临床样本进行深度测序和总miRNAs的评估，筛选microRNAs（miRNAs），用于结直肠癌的早期诊断。方法从正常结肠粘膜、结肠腺瘤和结直肠癌组织中提取总的小RNA，通过深度测序进行miRNA分析。然后通过生物信息学分析候选诊断性miRNA的测序数据，进一步验证其上调或下调状态。结果癌组织与正常粘膜比较，共鉴定出99个表达上调的miRNAs和90个表达下调的miRNAs。比较腺瘤和正常粘膜发现114个上调和107个下调的miRNA。比较癌和腺瘤组织发现70个上调和27个下调的miRNA。在12例癌症和息肉患者中验证了所选上调和下调的miRNA的表达。具体而言，对于上调的miRNA，与正常粘膜相比，miR-18 a-5 p和miR-21- 3 p在腺瘤和癌组织中显著上调;与正常粘膜相比，miR-135 b-5 p、miR-17- 5 p、miR-182- 5 p、miR-200 a-5 p和miR-200 c-3 p在癌组织中显著上调，但在腺瘤组织中的表达与正常粘膜相比差异无显著性。与正常粘膜相比，miR-183- 5 p和miR-96- 5 p在腺瘤组织中显著上调，但与正常粘膜相比，这些差异在癌组织中不显著。对于下调的miRNA，与正常粘膜相比，miR-133 a-3 p在腺瘤和癌组织中均显著下调;与正常粘膜相比，miR-204- 5 p、miR-125 b-5 p、miR-139- 5 p、miR-100- 5 p和miR-30 a-5 p在癌组织中显著下调，但与正常粘膜相比，它们在腺瘤组织中的差异表达不显著。结论小RNA深度测序法检测的部分miRNA可能对中国人结直肠癌的诊断和预后有重要意义。miR-18 a-5 p和miR-21- 3 p在腺瘤和癌组织中的表达上调或miR-133 a-3 p的表达下调可作为结直肠癌早期筛查的指标。其他miRNA，如miR-135 b-5 p、miR-17- 5 p、miR-182- 5 p、miR-200 a-5 p、miR-200 c-3 p、miR-183- 5 p和miR-96- 5 p，在癌组织中上调或下调，但在腺瘤组织中不上调或下调，在早期诊断中的意义有限。需要进一步研究以确定具有诊断价值的筛查指标。
Purpose This study aims to screen microRNAs (miRNAs), for an early diagnosis of colorectal cancer, by deep sequencing and evaluation of total miRNAs using clinical samples from a Chinese patient population. Methods Total small RNAs from normal colonic mucosa, colonic adenomas, and colorectal cancer tissues were prepared for miRNA analysis by deep sequencing. The sequencing data were then analyzed by bioinformatics for candidate diagnostic miRNAs, which were further validated for their up- or downregulation status. Results Comparison of cancer tissues with normal mucosa identified 99 upregulated and 90 downregulated miRNAs. Comparison of adenomas and normal mucosa found 114 upregulated and 107 downregulated miRNAs. Comparison of cancer and adenoma tissues found 70 upregulated and 27 downregulated miRNAs. Selected up- and downregulated miRNAs were validated for their expressions in 12 cases of patients with cancer and polyps. Specifically, for the upregulated miRNAs, miR-18a-5p and miR-21-3p were significantly upregulated in adenomas and cancer tissues, compared with the normal mucosa; miR-135b-5p, miR-17-5p, miR-182-5p, miR-200a-5p, and miR-200c-3p were significantly upregulated in cancer tissues compared to the normal mucosa, but their differential expression was not significant in adenoma tissues when compared with the normal mucosa. miR-183-5p and miR-96-5p were significantly upregulated in adenoma tissues when compared with normal mucosa, but these differences were not significant in cancer tissues when compared to normal mucosa. For the downregulated miRNAs, miR-133a-3p was significantly downregulated in both adenoma and cancer tissues when compared to normal mucosa; miR-204-5p, miR-125b-5p, miR-139-5p, miR-100-5p, and miR-30a-5p were significantly downregulated in cancer tissues compared to the normal mucosa, but their differential expression was not significant in adenoma tissue compared to normal mucosa. Conclusion The findings of this study show that a number of miRNAs might be important in the diagnosis and prognosis of colorectal cancer in Chinese patients using the method of small RNA deep sequencing. Upregulation of miR-18a-5p and miR-21-3p or downregulation of miR-133a-3p in adenoma and cancer tissues may serve as an index for early screening of colorectal cancer. Other miRNAs, such as miR-135b-5p, miR-17-5p, miR-182-5p, miR-200a-5p, miR-200c-3p, miR-183-5p, and miR-96-5p, which were either up- or downregulated, in cancer tissues, but not in adenoma tissues, have limited significance in early diagnosis. Further study is needed to determine a screening index with diagnostic value.