LRRTM1基因甲基化是胰腺癌的潜在诊断和预后标志物
LRRTM1 gene methylation is a potential diagnostic and prognostic marker of pan-creatic cancer
张帆 1张美英 2高嫒嫒 2郭明洲2
作者信息
- 1. 新乡医学院第三附属医院消化内科,河南 新乡 453003;中国人民解放军总医院第一医学中心消化内科医学部
- 2. 中国人民解放军总医院第一医学中心消化内科医学部
- 折叠
摘要
目的 探讨 LRRTM1 基因在胰腺癌中的表观遗传调控机制及其甲基化作为诊断和预后标志物的可能性.方法 应用半定量 RT-PCR与甲基化特异性 PCR(methylation-specific PCR,MSP)技术检测 6 株胰腺癌细胞系中 LRRTM1 的表达及启动子区甲基化情况,应用 MSP 在 101 例癌前病变样本和 277 例胰腺癌组织中检测 LRRTM1 启动子区甲基化情况,并分析 LRRTM1 甲基化与临床病理因素之间及胰腺癌患者总生存期的关系.结果 LRRTM1 在 Capan1、Panc3.11、AsPC1 和 PANC-28 细胞中缺失表达,其 MSP结果为完全甲基化状态,在 Capan2 和 CFpac1 中正常表达,MSP 结果为未甲基化状态.应用去甲基化药物 5-aza-dc 处理细胞后,LRRTM1 在 Capan1、Panc3.11、AsPC1 和 PANC-28 细胞恢复表达,在 Capan2 和 CFpac1 细胞中表达未改变.表明 LRRTM1 基因在胰腺癌中的表达受甲基化调控.在 16.83%(17/101)的癌前病变样本和 52.35%(145/277)的原发性胰腺癌中,LRRTM1 发生甲基化,且 LRRTM1 甲基化与患者的年龄相关(P<0.05).在获得随访信息的 232 例胰腺癌组织样本中发现 LRRTM1 甲基化与胰腺癌患者总生存期呈负相关.结论 LRRTM1 基因在胰腺癌组织中频繁发生甲基化,其表达受启动子区甲基化调控,LRRTM1 甲基化是胰腺癌潜在的诊断和预后标志物.
Abstract
Objective To investigate the epigenetic regulation of LRRTM1 gene expression and the possibility of LRRTM1 methylation as a diagnostic and prognostic marker.Methods LRRTM1 expression and promoter methylation status in 6 pancreatic cancer cell lines were detected by semi-quantitative RT-PCR and methylation-specific PCR(MSP).MSP was used to detect the methylation status of LRRTM1 in 101 cases of precancerous and 277 cases of pan-creatic cancer samples.Chi-square test was employed to analyze the association of LRRTM1 methylation and clinicopath-ological factors.Kaplan-Meier survival curves were plotted and Log-Rank test was used to compare overall survival.Uni-variate and multivariate Cox regression analyses were used to evaluate prognostic factors.Results The expression of LRRTM1 was lost in Capan1,Panc3.11,AsPC1 and PANC-28 cells,and the promoter region of LRRTM1 was com-pletely methylated in these cells.LRRTM1 was normally expressed in Capan2 and CFpac1 cells,and the promoter re-gion of LRRTM1 was unmethylated.The expression of LRRTM1 was restored in Capan1,Panc3.11,AsPC1 and PANC-28 cells,and unchanged in Capan2 and CFpac1 cells after 5-aza-dc treatment.These results indicated that the expression of LRRTM1 was regulated by promoter methylation in pancreatic cancer.It was further found that LRRTM1 was methylated in 16.83%(17/101)of precancerous samples and 52.35%(145/277)of primary pancreatic cancer,and the expression of LRRTM1 was associated with age(P<0.05).For 232 cases of pancreatic cancer patients with available survival data,LRRTM1 methylation was found to be significantly associated with poor overall survival(P<0.05).Conclusion LRRTM1 gene is frequently methylated in pancreatic cancer,and its expression is regulated by promoter methylation.LRRTM1 methylation is a potential diagnostic and prognostic marker for pancreatic cancer.
关键词
LRRTM1/胰腺癌/DNA甲基化/甲基化特异性PCRKey words
LRRTM1/Pancreatic cancer/DNA methylation/Methylation specific PCR引用本文复制引用
基金项目
国家重点研发计划(2020YFC2002705)
国家自然科学基金(82272632)
国家自然科学基金(81672318)
中国人民解放军总医院青年自主创新科学基金(22QNCZ027)
出版年
2024
NETL
NSTL
万方数据