Direct RT-qPCR for serum circulating Bmi-1 mRNA and its clinical application in colorectal cancer
作者单位：Department of Clinical LaboratoryQilu Hospital of Shandong University Department of Clinical LaboratoryThe Second Hospital of Shandong University
会议名称：《2017年中国肿瘤标志物学术大会暨第十一届肿瘤标志物青年科学家论坛》
会议日期：2017年
学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学]
摘 要：Purpose: Cell free Bmi-1 m RNA is stably detectable in the serum/plasma, and associated with development and progression of some tumors. Previous methods detecting extracellular Bmi-1 mRNA with RNA extraction are inefficient. This study developed a novel reverse transcription quantitative PCR(RTqPCR) approach directly applied in serum(RT-qPCR-D) to quantify Bmi-1 m RNA, and assessed its diagnostic and prognostic potential in colorectal cancer(CRC). Methods: The feasibility of RT-qPCR-D method was first analyzed in 50 serum samples. Then, using RT-qPCR-D method, Bmi-1 mRNA expression was validated in serum from an independent cohort of patients with 87 normal colonoscopy, 76 hyperplastic polyp, 82 inflammatory bowel disease, 68 adenoma and 158 CRC. Receiver operating characteristic(ROC) curves and Cox analyses were used to evaluate its diagnosis and prognosis value, respectively. Results: In pilot study, levels of Bmi-1 mRNA were increased in CRC serum samples detected by RT-qPCR-D, and significantly associated with results obtained by RT-qPCR. In validation cohort, serum Bmi-1 mRNA levels were significantly elevated in CRC group and adenoma group when compared with other groups. The area under ROC curve distinguishing CRC from benign colorectal diseases was 0.888 with 72.2% sensitivity and 94.9% specificity, which superior to CEA. Bmi-1 mRNA levels were significantly associated with survival. Cox analysis indicated Bmi-1 mRNA was an independent prognostic factor for overall survival. Conclusion: Detection of cell free Bmi-1 mRNA in serum by RT-qPCR-D is a simple and noninvasive approach, and may be used for CRC diagnosis and prognosis.