研究目的
To determine whether within primary CRCs with KRAS mutations intratumoral KRAS mutation heterogeneity can be detected between two strictly defined areas, i.e. the luminal (mucosa/submucosa) and peripheral invasive front of the tumor.
研究成果
No KRAS mutation heterogeneity was found between luminal and peripheral invasive parts of CRCs with KRAS mutations irrespective of tumor grade and site. This supports the idea that KRAS mutations are acquired early and remain during progression of CRC.
研究不足
The study was limited to the analysis of KRAS mutations in codons 12 and 13 and did not explore mutations in other codons or genes that may also influence resistance to anti-EGFR therapy.
1：Experimental Design and Method Selection:
Laser-capture microdissection was applied to separate tumor cells from benign stromal cells, and a commercially available high-sensitivity real-time PCR-based diagnostic assay for KRAS mutation testing, with the CE-IVD mark, was used for molecular analysis.
2：Sample Selection and Data Sources:
Tumor tissue from 14 FFPE CRCs in which KRAS mutations had been previously detected.
3：List of Experimental Equipment and Materials:
PALM MicroBeam laser-capture microdissection system, PNAClamp KRAS mutation detection kit, QIAamp DNA FFPE tissue kit, NanoDrop 2000 Spectrophotometer.
4：Experimental Procedures and Operational Workflow:
From every tumor about 400-500 nests of cancer cells were excised from each of the examined areas (luminal and peripheral) for molecular analysis.
5：Data Analysis Methods:
The presence of KRAS mutations was detected with the PNAClamp KRAS mutation detection kit.
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