乳腺癌患者外周血CK19 mRNA表达及不同化疗方案对其影响

Expression of peripheral blood CK19 mRNA in breast cancer and affection of different chemotherapy regimens

目的以CK19(cytokeratin-19)mRNA为基因标志检测不同乳腺癌患者外周血肿瘤细胞,检测化疗前后外周血有核细胞CK19mRNA表达量的改变及其与临床病理因素的关系,比较不同化疗方案下外周血有核细胞CK19mRNA表达量的变化。方法采用实时定量RT-PCR方法检测217例乳腺癌患者外周血CK19mRNA表达(17例导管内癌、176例浸润性导管癌和24例乳腺癌远处转移)。217患者中有67例完成了化疗全过程常规取血和查血,对这些患者的化疗前后外周血CK19mRNA拷贝数的改变及其与临床病理因素的关系进行分析。化疗方案为CEF方案(CEF组23例)、TEC方案(TEC组28例)和TP方案(TP组16例),3组均行化疗6个周期,21d为1个周期。由于217例患者拷贝数改变值在各组中的分布均不符合正态分布,统计学分析采用非参数秩和检验。结果在导管内癌组、浸润性导管癌组和乳腺癌转移组3组间,外周血CK19mRNA拷贝数差异有统计学意义(P=0.048),乳腺癌远处转移组外周血CK19mRNA拷贝中位数高于导管内癌组和浸润性导管癌组(P<0.050)。67例患者中,化疗前后外周血CK19mRNA拷贝数改变在临床分期组间差异有统计学意义(P=0.041),临床Ⅳ期患者化疗前后外周血CK19mRNA拷贝数改变的中位数高于Ⅰ～Ⅲ期患者(P<0.050)。PCNA阳性患者化疗前后外周血CK19mRNA拷贝数改变的中位数高于PCNA阴性患者,两组间差异有统计学意义(P=0.032)。肿瘤大小、淋巴结转移、组织学分级、ER、PR、以及HER-2亚组间化疗前后外周血CK19mRNA拷贝数改变的差异均无统计学意义(P>0.050)。3组患者化疗后外周血CK19mRNA拷贝数均有改变,3组间差异有统计学意义(P=0.011)。结论乳腺癌患者外周血CK19mRNA表达可以提示转移风险,TEC方案较TP和CEF方案对外周血CK19mRNA表达量改变的影响更显著,因此可以通过检测化疗前后外周血CK19mRNA表达量改变为术后化疗实施提供参考。

Objective To assay cancer cells in peripheral blood of breast cancer patient using CK19mRNA as gene marker and analyze the variation of CK19mRNA expression before and after chemotherapy and its relation with clinical pathologic factors, and compare the variation of CK19mRNA expression under different chemotherapy regimens. Methods RT-PCR was used to assay the expression of peripheral blood CK19mRNA in 217 breast cancer patients, including 176 cases of infiltrative ductal carcinoma （IDC group）, 17 in situ ductal carcinoma （DCIS group） and 24 metastatic breast cancer （MBC group）. Among the 217 patients, only 67 completed routine blood taking and examination dwing the whole process of chemotherapy. The variation of peripheral blood CK19mRNA copy number before and after chemotherapy and its relation with clinicopathological factors of these 67 patients were analyzed. Chemotherapy included CEF regime （Cyclophosphamide 600 mg/M^2, Epirubicin 90 mg/M^2,5 Fluorouracil 500 mg/M^2, CEF group n = 23）, TEC regime （Paclitaxel 140 mg/M^2, Epirubicin 90 mg/M^2 , Cyclophosphamide 600 mg/M^2 , TEC group n=28） and TP regime （Paclitaxel 140 mg/M^2 , Cis-platinum 60 mg/M^2 , TP group n=16）. There was a total of 6 cycles, 21 days as one cycle. Kruskal- Wallis test and Wilcoxon ran sum test were used for statistical analysis, for the distribution of CK19 mRNA copy numbers data of the 217 cases was not a normal distribution. Results Between the three groups of IDC group, DCIS group and MBC group, the peripheral blood CK19mRNA copy numbers were statistically different （P = 0. 048） ; the median number of the MBC group was higher than that of the other two groups （P〈0. 050）. For the 67 patients, the peripheral blood CK19mRNA copy number variation before and after chemotherapy was statistically different among subgroups of clinical stage Ⅰ - Ⅳ patients; the median number of the clinical stage Ⅳ patients was higher than that of stages Ⅰ - Ⅲ patients （P〈0. 050）. The median number of peripheral blood CK19mRNA copy number variation before and after chemotherapy was higher in PCNA positive patients than in PCNA negative patients, with statistical difference between the two （P= 0. 032）. The peripheral blood CK19mRNA copy number variations before and after chemotherapy in tumour size, lymph node metastasis, histological stage, ER, PR, and HER-2 had no statistical difference （P〉0. 050）. The peripheral blood CK19mRNA copy numbers after chemotherapy in the three chemotherapy groups all changed, there was a statistical difference between the three groups （P = 0. 011 ）. Conclusion The peripheral blood CK19mRNA expression of breast cancer can be used to assay distance metastatic status. TEC has stronger effect on CK19mRNA expression of peripheral blood compared with TP and CEF chemotherapy regimes, so the assessment of peripheral blood CK19mRNA expression can indirectly provide an reference for postoperative chemotherapy.