hMAM联合MMP-9和C-erbB2mRNA表达检测在乳腺癌外周血微转移诊断中的应用

目的：探讨生物标记物人乳腺珠蛋白（hMAM）联合基质金属蛋白酶9（MMP-9）和人表皮生长因子受体2（C-erbB2） mRNA在乳腺癌外周血微转移患者中阳性表达情况，阐明其用于诊断乳腺癌外周血微转移的临床应用价值。方法：选择74例乳腺癌患者、21例乳腺纤维腺瘤患者和10名健康人作为研究对象，所有患者均行手术治疗，采集外周血，运用实时荧光定量PCR检测外周血hMAM、MMP-9和C-erbB2 mRAN表达水平，比较hMAM、MMP-9和C-erbB2联合检测阳性表达率，并分析不同临床病理特征患者之间hMAM、MMP-9和C-erbB2联合检测的差异。结果：MMP-9、C-erbB2 mRNA阳性表达率在有无淋巴结转移患者间比较差异有统计学意义（χ²=6.450，P < 0.05；χ²=5.636，P < 0.05），hMAM mRNA阳性表达率在HER-2阳性与阴性患者间比较差异有统计学意义（χ²=5.804，P < 0.05）。乳腺癌患者hMAM及联合MMP-9和C-erbB2 mRNA阳性表达率分别为37.84%（28/74）、59.46%（44/74）和48.65%（36/74），三者联合阳性表达率为64.86%（48/74），均高于健康对照组（χ²=5.676，P < 0.05；χ²=3.102，P > 0.05；χ²=5.339，P < 0.05；χ²=2.310，P > 0.05）、乳腺纤维腺瘤组（χ²=8.438，P < 0.01；χ²=4.491，P < 0.05；χ²=7.982，P < 0.01；χ²=4.844，P < 0.05）和非乳腺癌组（对照组+乳腺纤维腺瘤组）（χ²=13.093，P < 0.01；χ²=6.471，P < 0.05；χ²=11.837，P < 0.01；χ²=6.103，P < 0.05）。与Ⅰ+Ⅱ期比较，Ⅲ+Ⅳ期乳腺患者hMAM mRNA单独及联合阳性表达率均增加，其中hMAM mRNA单独及联合C-erbB2 mRNA阳性表达率比较差异有统计学意义（χ²=5.157，P < 0.05；χ²=4.912，P < 0.05）。结论：hMAM诊断乳腺癌微转移的阳性率较低，联合MMP-9和C-erbB2 mRNA检测可以提高诊断阳性率，对于早期乳腺癌微转移诊断具有一定临床应用价值。

Application of detection of expressions of hMAM combined with MMP-9 and C-erbB2 mRNA in peripheral blood in diagnosis of micrometastases of breast cancer

Objective:To explore the positive expressions of biological markers human mammaglobin (hMAM) combined with matrix metallopeptidase 9 (MMP-9) and human epidermal growth factor receptor 2 (C-erbB2) mRNA in peripheral blood of the breast cancer patients with micrometastases,and to clarify its clinical application value in diagnosis of the micrometastases in peripheral blood of the breast cancer patients.Methods:A total of 74 patients with breast cancer,21 patients with breast fibroadenoma and 10 healthy controls were selected as the subjects.All the patients received surgical treatment and the peripheral blood was collected.The mRNA expression levels of hMAM,MMP-9 and C-erbB2 in peripheral blood were measured by the real-time fluorescent quantitative PCR.The positive expression rates of detection of hMAM,MMP-9 and C-erbB2 were compared,and the differences in detection of hMAM combined with MMP-9 and C-erbB2 between the patients with different clinicopathologic features were analyzed.Results:In the breast cancer patients with lymph node metastasis,the differences of positive expression rates of MMP-9 and C-erbB2 mRNA were significant (x2=6.450,P＜0.05;x2=5.636,P＜0.05),and the difference of positive expression rate of hMAM mRNA was sigificant between HER-2 positive and negative patients (x2=5.804,P＜0.05).The positive expression rates of individual hMAM and combined with MMP-9 and C-erbB2 were 37.84％ (28/74),59.46％ (44/74) and 48.65％ (36/74) in the breast cancer patients,the combined postive expression rate of these three kinds of markers was 64.86 ％ (48/74),which were higher than those in healthy controls group (x2=5.676,P＜0.05;x2=3.102,P＞0.05;x2=5.339,P＜0.05;x2 =2.310,P＞0.05),fibroadenoma of breast group (x2 =8.438,P＜0.01;x2 =4.491,P＜ 0.05;x2 =7.982,P＜0.01;x2 =4.844,P＜0.05) and non-breast cancer group (healthy controls group+ breast fibroadenoma group) (x2 =13.093,P＜0.01;xx2 =6.471,P＜0.05;x2 =11.837,P＜0.01;x2 =6.103,P＜ 0.05).The positive expression rates of individual hMAM and the joint detection in the breast cancer patients at stage Ⅲ + Ⅳ were higher than those in the patients at stage Ⅰ + Ⅱ;the positive expression rates of individual hMAM and combined with C-erbB2 were statistically significant (x2 =5.157,P＜0.05;x2 =4.912,P＜0.05).Conclusion:hMAM has a low positive rate in the diagnosis of micrometastases in the breast cancer patients,while hMAM combined with MMP-9 and C-erbB2 detection could improve the positive rates.which presents some clinical application value for the early diagnosis of breast cancer micrometastases.