血浆长链非编码RNA肺腺癌转移相关转录本1在乳腺癌中的表达及其临床意义

摘要

目的 观察乳腺癌患者血浆中长链非编码RNA(lncRNA)肺腺癌转移相关转录本1 (MALAT1)的表达变化,并探讨其临床意义.方法 应用实时荧光定量PCR方法评估MALAT1不同区域片段在10名健康人血浆中的表达,并检测102例术前、64例术后乳腺癌患者、47例良性乳腺肿瘤患者以及50名健康对照者血浆中内参甘油醛-3-磷酸脱氢酶(GAPDH)和MALAT1的表达水平.分析GAPDH的表达与健康人及患者各临床资料的关系,判断其稳定性.应用受试者工作特征曲线(ROC)分析MALAT1鉴别诊断乳腺癌的效能,并与CA153和CEA作比较.分析MALAT1的表达与患者临床病理特征之间的关系以及比较手术前后血浆中MALAT1的表达.正态分布计量资料采用t检验,多组间比较采用单因素方差分析,各组间MALAT1的相对表达量比较采用非参数秩和检验.结果 GAPDH在女性外周血中表达稳定,不受年龄和病理因素的影响(P>0.05),可以用作血浆lncRNAs检测的内源性参照.血浆MALAT1以片段形式存在,各片段表达差异有统计学意义(χ2=27.042,P<0.001).乳腺癌组术前血浆MALAT1的相对表达量5.58(2.17~12.34)高于良性组1.08 (0.61~2.58)(Z=6.209,P<0.001)及健康对照组1.63(0.98~3.51)(Z=4.871,P<0.001),而良性组与健康对照组之间,差异无统计学意义(Z=-1.675,P=0.094).Ⅰ-Ⅱ期乳腺癌患者术前血浆MALAT1较良性组异常高表达,差异有统计学意义(Z=5.593,P<0.001).术后血浆MALAT1相对表达与术前相比降低(Z=-2.248,P=0.025).ROC曲线分析结果显示MALAT1、CA153和CEA曲线下面积AUC分别为0.744、0.619和0.553;三者敏感度分别为54.1%、60.0%、70.0%;特异度分别为86.3%、66.7%、44.1%.乳腺癌患者血浆MALAT1的表达与TNM分期(Z=-1.982,P=0.047)、淋巴结转移(Z=-2.186,P=0.029)和病理分化程度(Z=-2.435,P=0.015)相关.结论 乳腺癌患者血浆中MALAT1高表达,可能是乳腺癌辅助诊断的一项潜在检测指标.%Objective To investigate the level of plasma MALAT1 in breast cancer(BC)patients and its clinical significance.Methods The expression levels of MALAT1 different fragments were detected in plasma from 10 healthy controls.The expressions of GAPDH and MALAT1 of plasma samples collected from 102 preoperative breast cancer patients,64 postoperative breast cancer patients,47 breast benign tumor patients and 50 healthy controls were determined by RT-qPCR.The potential association between plasma GAPDH level and cases′clinicopathologic features was analyzed to evaluate the stability of GAPDH. Receiver operating characteristic(ROC)curve was constructed to evaluate the diagnostic efficiency of MALAT1,CA153 and CEA for breast cancer.Meanwhile, the association between MALAT1 level and the clinicopathologic features and the expressions of MALAT 1 between preoperative and postoperative BC patients were analyzed.T-test and one-factor ANOVA test were used for normal distribution of quantitative data.The rank sum test was used for non-normal distribution of data.Results GAPDH level was stable in female plasma and was not affected by age and pathology(P>0.05).GAPDH can be used as a reference for plasma lncRNA detection.The levels of different MALAT1 fragments were inconsistent(χ2=27.042,P<0.001).Levels of MALAT1 were significantly elevated in preoperative BC patients[5.58(2.17-12.34)] compared with breast benign tumor patients and healthy controls[1.08(0.61 -2.58)(Z=6.209,P<0.001),1.63(0.98 -3.51)(Z=4.871,P<0.001)].However, there was no significantly difference between breast benign tumor patients and healthy controls(Z=-1.675,P=0.094).The MALAT1 levels of low grade patients(gradeI and II)were higher than those of breast benign tumor patients(Z=5.593,P<0.001).The relative expression of MALAT1 in postoperative plasma was significantly reduced(Z=-2.248,P=0.025).Areas under the ROC curve of MALAT1,CA153 and CEA were 0.744,0.619 and 0.553 respectively.The sensitivity and specificity were 54.1%,60.0%,70.0% and 86.3%,66.7%, 44.1%respectively.The levels of MALAT1 were associated with TNM stage(Z=-1.982,P=0.047), lymph node metastasis(Z=-2.186,P=0.029)and tumor differentiation(Z=-2.435,P=0.015). Conclusion The expressions of MALAT1 were highly elevated in BC patients.Plasma MALAT1 may be an important biomarker for the diagnosis of breast cancer.