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结蛋白(CLU)mRNA 在结直肠癌患者肿瘤中的高表达水平预示着预后不良。

High clusterin (CLU) mRNA expression levels in tumors of colorectal cancer patients predict a poor prognostic outcome.

机构信息

Department of Biochemistry and Molecular Biology, Faculty of Biology, National and Kapodistrian University of Athens, Athens GR-15701, Greece.

Department of Cell Biology and Biophysics, Faculty of Biology, National and Kapodistrian University of Athens, Athens GR-15784, Greece.

出版信息

Clin Biochem. 2020 Jan;75:62-69. doi: 10.1016/j.clinbiochem.2019.10.008. Epub 2019 Oct 28.

Abstract

OBJECTIVES

Clusterin (CLU) is a multifunctional intra-/extra-cellular molecular chaperone with indications of serving as a promising prognostic biomarker for colorectal cancer (CRC). Several studies have examined the potential prognostic value of the CLU protein in CRC; however, our research follows an alternative approach, focusing on the CLU mRNA expression.

DESIGN AND METHODS

Total RNA from 172 cancerous tissue specimens and 39 paired non-cancerous ones was isolated and 2 μg of this were subjected to reverse transcription with an oligo-dT primer. The single stranded DNA, which was synthesized, was amplified with an in-house developed highly sensitive and precise qPCR method, using specific pair of primers for the CLU molecule. Finally, an extensive biostatistical analysis took place for the assessment of the results.

RESULTS

Patients with tumors expressing high CLU mRNA levels had a higher probability for poor outcome (relapse and death), comparing to those with CLU mRNA-negative tumors. This association between CLU mRNA expression status and both disease-free survival (DFS) and overall survival (OS) is evident in Cox regression analysis and is also depicted in the Kaplan-Meier survival curves. Consistently, the aforementioned associations and the CLU mRNA expression levels are significantly enhanced as CRC tumors progress from TNM stage I to IV, further supporting the functional implication of CLU in tumorigenesis.

CONCLUSIONS

High CLU mRNA levels in CRC tumors can act as a new adverse prognostic biomarker of DFS and OS for CRC, independent of clinicopathological and biological features of the patient.

摘要

目的

簇集蛋白(CLU)是一种多功能的细胞内/细胞外分子伴侣,有迹象表明它可以作为结直肠癌(CRC)有前途的预后生物标志物。有几项研究检查了 CLU 蛋白在 CRC 中的潜在预后价值;然而,我们的研究采用了一种替代方法,重点研究 CLU mRNA 的表达。

设计和方法

从 172 个癌组织标本和 39 对非癌组织中分离总 RNA,并将 2μg 总 RNA 用 oligo-dT 引物进行逆转录。合成的单链 DNA 用我们开发的高灵敏度和精确的 qPCR 方法进行扩增,该方法使用 CLU 分子的特异性引物。最后,进行了广泛的生物统计学分析以评估结果。

结果

与 CLU mRNA 阴性肿瘤相比,表达高 CLU mRNA 水平的肿瘤患者发生不良结局(复发和死亡)的可能性更高。CLU mRNA 表达状态与无病生存期(DFS)和总生存期(OS)之间的这种关联在 Cox 回归分析中很明显,在 Kaplan-Meier 生存曲线中也有描述。一致地,上述关联和 CLU mRNA 表达水平随着 CRC 肿瘤从 TNM 分期 I 期进展到 IV 期而显著增强,进一步支持 CLU 在肿瘤发生中的功能意义。

结论

CRC 肿瘤中高 CLU mRNA 水平可作为 CRC 的 DFS 和 OS 的新不良预后生物标志物,独立于患者的临床病理和生物学特征。

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