外周血RUNX2基因甲基化及表达水平与乳腺癌新辅助化疗敏感性的相关性研究

Correlation between methylation and expression of RUNX2 gene in peripheral blood and sensitivity to neoadjuvant chemotherapy in breast cancer

目的探讨外周血Runt相关转录因子2(Runt associated transcription factor 2,RUNX2)基因甲基化及表达水平与乳腺癌新辅助化疗敏感性的相关性。方法纳入2021年1月至2022年6月于济宁医学院附属医院首次接受全程新辅助化疗并进行手术切除治疗的90例女性乳腺癌患者作为研究对象,同时纳入同期于我院进行体检的90名健康女性作为对照组。采用亚硫酸盐转化法检测所有研究对象的外周血RUNX2基因甲基化水平,采用荧光定量PCR反应检测所有研究对象的外周血RUNX2基因mRNA表达水平。将乳腺癌患者经新辅助化疗后的效果分为敏感和不敏感组。结果乳腺癌患者外周血RUNX2基因甲基化阳性率为41.11%(37/90),显著低于对照组64.44%(58/90),差异有统计学意义(χ^(2)=9.83,P=0.002);乳腺癌患者的外周血RUNX2基因mRNA表达水平为1.73±0.24,显著高于对照组1.19±0.18,差异有统计学意义(t=5.22,P<0.001)。乳腺癌患者外周血RUNX2基因甲基化阳性患者的基因mRNA表达水平为1.26±0.20,显著低于RUNX2基因甲基化阴性患者1.84±0.26,差异有统计学意义(t=4.98,P<0.001)。乳腺癌新辅助化疗敏感者的外周血RUNX2基因甲基化阳性率为49.28%(34/69),显著高于不敏感者14.29%(3/21),差异有统计学意义(χ^(2)=8.14,P=0.004)。新辅助化疗敏感者的外周血RUNX2基因mRNA表达水平为1.62±0.17,显著低于不敏感者2.09±0.22,差异有统计学意义(t=5.28,P<0.001)。ROC曲线结果显示外周血RUNX2基因mRNA表达水平预测乳腺癌新辅助化疗效果的灵敏度和特异度分别为85.7%和85.5%。结论外周血RUNX2基因甲基化及表达水平与乳腺癌新辅助化疗敏感性有关,可作为预测乳腺癌新辅助化疗疗效的标志物。

Objective To investigate the correlation between the methylation and expression of Runt associated transcription factor 2(RUNX2)gene in peripheral blood and sensitivity to neoadjuvant chemotherapy in breast cancer.Methods 90 female breast cancer patients who received full-course neoadjuvant chemotherapy and surgical resection for the first time in our hospital from Jan.2021 to Jun.2022 were included as the study objects,and 90 healthy women who underwent physical examination in our hospital during the same period were included as the control group.The methylation level of RUNX2 gene in peripheral blood of all subjects was detected by sulfite conversion method,and the mRNA expression level of RUNX2 gene in peripheral blood was detected by fluorescence quantitative PCR.The effects of neoadjuvant chemotherapy in breast cancer patients were divided into sensitive and insensitive groups.Results The positive rate of RUNX2 gene methylation in peripheral blood of breast cancer patients was 41.11%(37/90),significantly lower than that of the control group(64.44%(58/90)),the difference was statistically significant(χ^(2)=9.830,P=0.002).The mRNA expression level of RUNX2 gene in peripheral blood of breast cancer patients was 1.73±0.24,which was significantly higher than that of the control group(1.19±0.18),and the difference was statistically significant(t=5.221,P<0.001).The mRNA expression level of RUNX2 methylation-positive patients in peripheral blood of breast cancer patients was 1.26±0.20,significantly lower than that of RUNX2 methylation-negative patients(1.84±0.26),and the difference was statistically significant(t=4.981,P<0.001).The positive rate of RUNX2 gene methylation was 49.28%(34/69)in patients with sensitivity to neoadjuvant chemotherapy,significantly higher than 14.29%(3/21)in patients with insensitive chemotherapy,and the difference was statistically significant(χ^(2)=8.142,P=0.004).The mRNA expression level of RUNX2 gene in peripheral blood of neoadjuvant chemotherapy sensitive patients was 1.62±0.17,which was significantly lower than that of insensitive patients(2.09±0.22),and the difference was statistically significant(t=5.283,P<0.001).The ROC curve showed that the sensitivity and specificity of RUNX2 mRNA expression in peripheral blood to predict the effect of neoadjuvant chemotherapy were 85.7%and 85.5%,respectively.Conclusion The methylation and expression level of RUNX2 gene in peripheral blood are related to the sensitivity of neoadjuvant chemotherapy in breast cancer,and can be used as a marker to predict the efficacy of neoadjuvant chemotherapy in breast cancer.