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通过液体活检比较结直肠癌患者循环肿瘤 DNA 水平与组织学分级的 KRAS 基因突变。

Comparison of KRAS gene in circulating tumor DNA levels vs histological grading of colorectal cancer patients through liquid biopsy.

机构信息

Department of Biochemistry, Basic Health Sciences, Ziauddin University, Karachi, Pakistan.

Department of Pharmacology, Basic Health Sciences, Ziauddin University, Karachi, Pakistan.

出版信息

Saudi J Gastroenterol. 2023 Nov-Dec;29(6):371-375. doi: 10.4103/sjg.sjg_85_23.

Abstract

BACKGROUND

To determine KRAS gene in circulating tumor DNA in comparison with histological grading through liquid biopsy in colorectal cancer patients.

METHODS

This dual-centered cross-sectional study included 73 diagnosed patients of colorectal cancer at different grading levels [Grade I, well differentiated (n = 7, 9.5%); Grade II, moderately differentiated (n = 14,18.9%); and Grade III, poorly differentiated (n = 52, 70%)]. Blood was collected, and plasma was separated. ctDNA was extracted, using magnetic bead-based technique (MagMAX Cell-Free DNA kit). KRAS gene was quantified through qPCR. STRING database was used to find KRAS interactomes.

RESULTS

Mean threshold cycle (CT value) of KRAS gene in Grade III samples showed significantly higher (P = 0.001) levels of ctDNA (2.7 ± 1.14) compared with Grade II and Grade I (3.1 ± 0.68, 2.3 ± 0.60), respectively. Grading characterization showed that rectal cancer (n = 22, 42.3%) with Grade III (68.8%) was more prevalent than colon and sigmoid cancer (n = 19, 36.5%, n = 11, 21%, respectively). STRING database showed 10 functional genes interacting with KRAS expressed as gene/proteins.

CONCLUSION

Liquid biopsy can be used to detect ctDNA in plasma of CRC patients and enabled to detect the KRAS gene by qPCR. The technique being less invasive and cost-effective is convenient for multiple biopsies in different cancers.

摘要

背景

通过液体活检在结直肠癌患者中比较循环肿瘤 DNA 中的 KRAS 基因与组织学分级。

方法

本双中心横断面研究纳入了不同分级水平的 73 例结直肠癌确诊患者[I 级,分化良好(n=7,9.5%);II 级,中度分化(n=14,18.9%);III 级,分化不良(n=52,70%)]。采集血液,分离血浆。采用基于磁珠的技术(MagMAX Cell-Free DNA 试剂盒)提取 ctDNA。通过 qPCR 定量 KRAS 基因。使用 STRING 数据库寻找 KRAS 相互作用体。

结果

III 级样本中 KRAS 基因的平均阈值循环(CT 值)显示 ctDNA 水平明显更高(P=0.001)(2.7±1.14),与 II 级和 I 级相比(3.1±0.68,2.3±0.60)。分级特征显示,III 级(68.8%)的直肠癌(n=22,42.3%)比结肠癌和乙状结肠癌更常见(n=19,36.5%;n=11,21%)。STRING 数据库显示 10 个与 KRAS 相互作用的功能基因表达为基因/蛋白。

结论

液体活检可用于检测 CRC 患者血浆中的 ctDNA,并通过 qPCR 检测 KRAS 基因。该技术具有微创和经济有效的特点,便于在不同癌症中进行多次活检。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1c53/10754382/f2eaaad5f271/SJG-29-371-g001.jpg

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