血清 miR-133b 和 miR-206 的下调与临床进展结果相关,可作为监测转移性结直肠癌患者的生物标记物

Surasak Wanram, Namphon Klaewkla, Parichart Pinyosri
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引用次数: 0

摘要

背景:结直肠癌(CRC)是全球第三大常见癌症。非编码 RNA 或微 RNA(miRNA;miRs)生物标志物可在癌症发生和发展过程中发挥作用。特定的 KRAS 和表皮生长因子受体(EGFR)突变与 CRC 的发展有关,在控制细胞过程中扮演着表观遗传学事件的角色。循环血清 miRs 可作为生物标志物用于 CRC 的早期诊断、监测和预后,但在临床上仍不明确:目的:确定 CRC 患者循环血清 miR-133b 和 miR-206 的潜在生物标志物:采用 TargetScanHu-man7.2、miRTar2GO、miRDB、MiRanda 和 DIANA-microT-CDS 对 microRNA 进行生物信息学预测。随后提取了 44 份 CRC 血清样本(19 份局部晚期 CRC、23 份远处晚期 CRC)和 12 份正常血清样本进行 RNA 分离、cDNA 合成和 miR 验证。通过定量 RT-PCR 验证了候选循环血清 miR-133b 和 miR-206 的相对表达。用 2-ΔΔCt 方法将相对表达归一化为尖峰内对照,并与正常样本进行比较,原则上以 1 为标准:结果表明,miR-206、miR-155-5p、miR-143-3p、miR-193a-3p、miR-30a-5p、miR-30d-5p、miR-30e-5p、miR-543、miR-877-5p 中的 9 个 miRs 与 KRAS 特异性 miRs 有关,而 miR-132-3p 中的 9 个 miRs 与 KRAS 特异性 miRs 有关、通过使用生物信息学预测工具,miR-133b、miR-302a-3p、miR-302b-3p、miR-302d-3p、miR-302e、miR-520a-3p、miR-520b、miR-520c-3p 和 miR-7-5p 等 9 个 miR 与表皮生长因子受体特异性 miR 相关。我们的研究结果表明,与正常人相比,循环血清中 miR-133b 和 miR-206 的表达水平明显降低,这与 CRC 患者(局部和晚期转移)有关(P < 0.05):循环血清miR-133b和miR-206可作为重要的生物标志物,用于监测转移性CRC患者的临床预后。临床上应进一步探讨与关键分子干预相关的药物反应性 CRC 患者的增加。
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Downregulation of Serum miR-133b and miR-206 Associate with Clinical Outcomes of Progression as Monitoring Biomarkers for Metastasis Colorectal Cancer Patients.

Background: Colorectal cancer (CRC) is the third most common cancer in the world. Noncoding RNAs or microRNAs (miRNAs; miRs) biomarkers can play a role in cancer carcinogenesis and progression. Specific KRAS and EGFR mutation are associated with CRC development playing a role in controlling the cellular process as epigenetic events. Circulating serum miRs can serve for early diagnosis, monitoring, and prognosis of CRC as biomarkers but it is still unclear, clinically.

Objective: To determine potential biomarkers of circulating serum miR-133b and miR-206 in CRC patients Methods: Bioinformatic prediction of microRNA was screened followed by TargetScanHuman7.2, miRTar2GO, miRDB, MiRanda, and DIANA-microT-CDS. Forty-four CRC serum (19 locally advanced, 23 distant advanced CRC) and 12 normal serum samples were subsequently extracted for RNA isolation, cDNA synthesis, and miR validation. The candidate circulating serum miR-133b and miR-206 were validated resulting in a relative expression via quantitative RT-PCR. Relative expression was normalized to the spike-internal control and compared to normal samples as 1 using the -2ΔΔCt method in principle.

Results: Our results represented 9 miRs of miR-206, miR-155-5p, miR-143-3p, miR-193a-3p, miR-30a- 5p, miR-30d-5p, miR-30e-5p, miR-543, miR-877-5p relate to KRAS-specific miRs, whereas, 9 miRs of miR-133b, miR-302a-3p, miR-302b-3p, miR-302d-3p, miR-302e, miR-520a-3p, miR-520b, miR-520c- 3p and miR-7-5p relevance to EGFR-specific miRs by using the bioinformatic prediction tools. Our results showed a decreased expression level of circulating serum miR-133b as well as miR-206 associating with CRC patients (local and advanced metastasis) when compared to normal (P < 0.05), significantly.

Conclusion: The circulating serum miR-133b and miR-206 can serve as significant biomarkers for monitoring the clinical outcome of progression with metastatic CRC patients. Increased drug-responsive CRC patients associated with crucial molecular intervention should be further explored, clinically.

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