Background: Colorectal cancer (CRC) remains a major global health problem, with rising incidence among younger individuals. The implementation of next-generation sequencing (NGS) has enabled comprehensive multigene analysis to identify cancer-predisposing variants and molecular alterations in tumors. However, data on age-related genetic differences in CRC from Central and Eastern European populations, including Poland, remain limited. Methods: This study aimed to explore molecular differences in CRC between patients aged ≤50 and >50 years in a Polish cohort. Tumor DNA was extracted from formalin-fixed, paraffin-embedded (FFPE) tissue samples obtained from 54 treatment-naive patients. Targeted sequencing of hot spot regions of 50 genes with known association to cancer was performed using an AmpliSeq for Illumina Cancer Hotspot Panel v2. Results: Variant frequencies in younger vs. older patients were:TP53(71.4% vs. 57.6%),APC(57.1% vs. 45.5%),KRAS(28.1% vs. 72.7%),NRAS(28.6% vs. 0%),SMAD4(9.5% vs. 12.1%),PIK3CA(14.3% vs. 24.2%), andFBXW7(4.8% vs. 14.7%). Co-occurrence ofAPC/KRAS/TP53variants was observed in 20% of cases.KRASmutations were significantly more frequent in older patients (p-value = 0.001), whileNRASmutations occurred exclusively in younger patients (29% vs. 0%,p= 0.021). Overall, 46% of patients exhibited multiple gene alterations (≥3 mutations). Notably,IDH1andCTNNB1variants were found only in patients with better prognosis, whereasTP53variants were nearly five times more frequent in patients with worse outcomes. Conclusions: Multigene panel sequencing revealed distinct age-related molecular patterns in CRC. Younger patients were more likely to harborNRASvariants, whereasKRASalterations predominated in older individuals. These findings underscore the relevance of NGS-based multigene profiling for risk stratification and personalized therapy in colorectal cancer.
背景：结直肠癌（CRC）仍是全球重大健康问题，其在年轻人群中的发病率呈上升趋势。新一代测序（NGS）技术的应用使得通过全面多基因分析识别癌症易感变异和肿瘤分子改变成为可能。然而，包括波兰在内的中欧和东欧人群结直肠癌年龄相关遗传差异的数据仍然有限。方法：本研究旨在探讨波兰队列中年龄≤50岁与>50岁患者结直肠癌的分子差异。从54例初治患者的福尔马林固定石蜡包埋（FFPE）组织样本中提取肿瘤DNA，使用Illumina癌症热点Panel v2扩增子测序技术对50个已知癌症相关基因的热点区域进行靶向测序。结果：年轻与年长患者的变异频率分别为：TP53（71.4% vs. 57.6%）、APC（57.1% vs. 45.5%）、KRAS（28.1% vs. 72.7%）、NRAS（28.6% vs. 0%）、SMAD4（9.5% vs. 12.1%）、PIK3CA（14.3% vs. 24.2%）及FBXW7（4.8% vs. 14.7%）。20%的病例中观察到APC/KRAS/TP53变异共存现象。KRAS突变在年长患者中显著更常见（p值=0.001），而NRAS突变仅出现在年轻患者中（29% vs. 0%，p=0.021）。总体而言，46%的患者存在多重基因改变（≥3个突变）。值得注意的是，IDH1和CTNNB1变异仅见于预后较好的患者，而TP53变异在预后较差患者中的出现频率高出近五倍。结论：多基因Panel测序揭示了结直肠癌中显著的年龄相关分子模式。年轻患者更易携带NRAS变异，而KRAS改变在年长个体中占主导地位。这些发现强调了基于NGS的多基因分析在结直肠癌风险分层和个体化治疗中的重要意义。
Age-Related Multigene Analysis of Colorectal Cancer Using Next-Generation Sequencing
肿瘤(癌症)患者之家