收稿日期: 2024-05-23
网络出版日期: 2024-10-22
基金资助
北京市重点专科建设项目(京卫医［2021］135号)
Methylation levels of the HIST1H4F gene mRNA region DNA and its diagnostic value in lung cancer tissues
Received date: 2024-05-23
Online published: 2024-10-22
目的 探讨H4成簇组蛋白6（HIST1H4F）基因mRNA区6个CpG位点的DNA甲基化水平及其在肺癌组织中的诊断价值。 方法 15例肺癌及癌旁配对正常肺组织的DNA甲基化水平采用焦磷酸测序检测。在初步评价基础上，建立甲基化敏感的限制性内切酶-荧光定量PCR（MSRE-qPCR）方法，分别检测测试组（肺腺癌60例、鳞癌38例、良性疾病30例和正常肺组织26例）及验证组（肺腺癌36例、鳞癌16例、良性疾病21例和正常肺组织23例）的DNA甲基化水平，采用ROC曲线评价其诊断价值。 结果 焦磷酸测序结果显示肺癌甲基化水平显著高于癌旁配对正常肺组织（P < 0.005）。MSRE-qPCR检测结果显示，测试组和验证组诊断肺癌的ROC曲线下面积分别为0.894和0.888，敏感度为76.5%和73.1%，特异度为92.9%和97.7%。在肺癌患者中，甲基化水平与吸烟显著相关（r = 0.273， P < 0.01）。 结论 HIST1H4F基因mRNA区的6个CpG位点可作为肺癌诊断的生物标记物，为临床肺癌诊断提供了新的分子靶标。
朱玲 , 任德旺 , 马润阳 , 梁国威 , 窦学军 . HIST1H4F基因mRNA区DNA甲基化水平及其在肺癌组织中的诊断价值[J]. 实用医学杂志, 2024 , 40(19) : 2726 -2732 . DOI: 10.3969/j.issn.1006-5725.2024.19.011
Objective To evaluate the methylation levels of DNA at six specific CpG sites located in the mRNA region of histone cluster 4 subfamily F member 6 （HIST1H4F） gene and determine their diagnostic significance about lung cancer. Methods The DNA methylation levels of 15 cases of lung cancer and adjacent paired normal lung tissue were detected using pyrophosphate sequencing. Based on preliminary evaluation， a methylation-sensitive restriction enzyme-fluorescence quantitative PCR （MSRE-qPCR） method was developed to detect DNA methylation levels in the test group （60 cases of lung adenocarcinoma， 38 cases of squamous cell carcinoma， 30 cases of benign diseases， and 26 cases of normal lung tissue） and the validation group （36 cases of lung adenocarcinoma， 16 cases of squamous cell carcinoma， 21 cases of benign diseases， and 23 cases of normal lung tissue）. The diagnostic value was evaluated using ROC curves. Results The results of pyrophosphate sequencing showed that the methylation levels of lung cancer were significantly higher than that of paired normal lung tissue （P < 0.005）. The detection results of MSRE-qPCR showed that the areas under the ROC curve for diagnosing lung cancer in the test group and validation group were 0.894 and 0.888， with sensitivity of 76.5% and 73.1%， and specificity of 92.9% and 97.7%， respectively. The methylation levels were significantly positively correlated with smoking in lung cancer patients （r = 0.273， P < 0.01）. Conclusion The six CpG sites in the mRNA region of the HIST1H4F gene can serve as biomarkers for diagnosing lung cancer， providinga new molecular target for clinical lung cancer diagnosis.
Key words： HIST1H4F; methylation; lung cancer; tissue; diagnosis
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