Chang Kun, Kong Yun-Yi, Dai Bo, Ye Ding-Wei, Qu Yuan-Yuan, Wang Yue, Jia Zhong-Wei, Li Gao-Xiang
Department of Urology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China.
Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China.
Oncotarget. 2015 Dec 8;6(39):41825-36. doi: 10.18632/oncotarget.6167.
Although circulating tumor cell (CTC) enumeration in peripheral blood has already been validated as a reliable biomarker in predicting prognosis in metastatic castration-resistant prostate cancer (mCRPC), patients with favorable CTC counts (CTC < 5/7.5 ml) still experience various survival times. Assays that can reduce patients' risks are urgently needed. In this study, we set up a real-time quantitative polymerase chain reaction (RT-qPCR) method to detect epithelial-mesenchymal transition (EMT) and stem cell gene expression status in peripheral blood to validate whether they could complement CTC enumeration. From January 2013 to June 2014 we collected peripheral blood from 70 mCRPC patients and enumerated CTC in these blood samples using CellSearch system. At the same time, stem cell-related genes (ABCG2, PROM1 and PSCA) and EMT-related genes (TWIST1 and vimentin) were detected in these peripheral blood samples using an RT-qPCR assay. Patient overall survival (OS) and treatment methods were recorded in the follow-up. For patients who received first-line chemotherapy, docetaxel plus prednisone, PSA progression-free survival (PSA-PFS) and PSA response rate were recorded. At the time of analysis, 35 patients had died of prostate cancer with a median follow-up of 16.0 months. Unfavorable CTC enumerations (CTC ≥5/7.5 ml) were predictive of shorter OS (p = 0.01). Also, positive stem cell gene expression indicated poor prognosis in mCRPC patients (p = 0.01). However, EMT gene expression status failed to show any prognostic value in OS (p = 0.78). A multivariate analysis indicated that serum albumin (p = 0.04), ECOG performance status (p < 0.01), CTC enumeration (p = 0.02) and stem cell gene expression status (p = 0.01) were independent prognostic factors for OS. For the 40 patients categorized into the favorable CTC enumeration group, positive stem cell gene expression also suggested poor prognosis (p < 0.01). A combined prognostic model consisting of stem cell gene expression and CTC enumeration increased the concordance probability estimated value from 0.716 to 0.889 in comparison with CTC enumeration alone. For patients who received docetaxel plus prednisone as first-line chemotherapy, positive stem cell gene expression suggested a poor PSA-PFS (p = 0.01) and a low PSA response rate (p = 0.008). However, CTC enumeration and EMT gene expression status did not affect PSA-PFS or PSA response rates. As a result, detection of peripheral blood stem cell gene expression could complement CTC enumeration in predicting OS and docetaxel-based treatment effects in mCRPC patients.
尽管外周血循环肿瘤细胞(CTC)计数已被确认为预测转移性去势抵抗性前列腺癌(mCRPC)预后的可靠生物标志物,但CTC计数良好(CTC<5/7.5 ml)的患者仍有不同的生存时间。迫切需要能够降低患者风险的检测方法。在本研究中,我们建立了一种实时定量聚合酶链反应(RT-qPCR)方法来检测外周血中上皮-间质转化(EMT)和干细胞基因表达状态,以验证它们是否可以补充CTC计数。2013年1月至2014年6月,我们收集了70例mCRPC患者的外周血,并使用CellSearch系统对这些血样中的CTC进行计数。同时,使用RT-qPCR检测这些外周血样本中的干细胞相关基因(ABCG2、PROM1和PSCA)和EMT相关基因(TWIST1和波形蛋白)。随访中记录患者的总生存期(OS)和治疗方法。对于接受一线化疗(多西他赛加泼尼松)的患者,记录前列腺特异抗原无进展生存期(PSA-PFS)和PSA缓解率。在分析时,35例患者死于前列腺癌,中位随访时间为16.0个月。不利的CTC计数(CTC≥5/7.5 ml)预示着较短的OS(p=0.01)。此外,干细胞基因表达阳性表明mCRPC患者预后不良(p=0.01)。然而,EMT基因表达状态在OS中未显示出任何预后价值(p=0.78)。多因素分析表明,血清白蛋白(p=0.04)、美国东部肿瘤协作组(ECOG)体能状态(p<0.01)、CTC计数(p=0.02)和干细胞基因表达状态(p=0.01)是OS的独立预后因素。对于40例CTC计数良好的患者,干细胞基因表达阳性也提示预后不良(p<0.01)。与单独的CTC计数相比,由干细胞基因表达和CTC计数组成的联合预后模型将一致性概率估计值从