Detecting carcinoma cells in peripheral blood of patients with hepatocellular carcinoma by Immunomagnetic beads and RT-PCR

	Detecting carcinoma cells in peripheral blood of patients with hepatocellular carcinoma by Immunomagnetic beads and RT-PCR
	Guo, JM; Yao, F; Lou, YL; Xu, CF; Xiao, BX; Zhou, WH; Chen, J; Hu, YR; Liu, Z
刊名	JOURNAL OF CLINICAL GASTROENTEROLOGY
	2007
卷号	41 期号:8 页码:783-788
关键词	liver cancer metastasis AFP mRNA circulating tumor cells
通讯作者	Guo, JM (reprint author), Ningbo Univ Sch Med, Ningbo 315211, Zhejiang, Peoples R China.,junmingguo@yahoo.com
英文摘要	Background: Increasing the sensitivity and specificity of detecting circulating carcinoma cells of patients with hepatocellular carcinoma (HCC) is very important for monitoring recurrence. Goal: To establish a novel method of detecting circulating carcinoma cells. Study: For method development, 3 sets of controls using HCC cell line HepG2 cells were used. (A): Serial dilutions of HepG2 cells were directly used to extract total RNA for nested reverse transcription-polymerase chain reaction (RT-PCR). (B): Five milliliter of healthy blood was spiked with a serial dilution of HepG2 cells and was used for Ficoll density gradient centrifugation to recover cells. The cells were used to extract total RNA for RT-PCR. (Q: After cell recovery with the same procedure as B, the cells were sorted sequentially by CD45 and Ber-EP4 immunomagnetic beads and used for RNA extraction and RT-PCR. For clinical samples, 44 patients with HCC and 7 healthy subjects were included. The alpha-fetoprotein mRNA was amplified using nested RT-PCR technique. Results: The spiking experiments using HepG2 cells showed that 10 cells in 5 mL blood could be detected by method C and an excellent dose-response to the number of spiked cells. Whereas, method B lacked any dose-response and would yield high false-positive rates. In clinical samples, the improved method led to a positive detection rate of 52.9%, 76.9%, and 92.9% in Child-Plug class A. B, and C. respectively. There was significant difference between class A and class C (P < 0.05). The total positive detection rate was 72.7%.
学科主题	Gastroenterology & Hepatology
类目[WOS]	Gastroenterology & Hepatology
关键词[WOS]	FETOPROTEIN MESSENGER-RNA ; ALPHA-FETOPROTEIN ; TUMOR-CELLS ; LIVER RESECTION ; GENE-EXPRESSION ; BONE-MARROW ; CANCER ; RECURRENCE ; DISEASE ; DISSEMINATION
收录类别	SCI
语种	英语
WOS记录号	WOS:000249135000010
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1586]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Guo, JM,Yao, F,Lou, YL,et al. Detecting carcinoma cells in peripheral blood of patients with hepatocellular carcinoma by Immunomagnetic beads and RT-PCR[J]. JOURNAL OF CLINICAL GASTROENTEROLOGY,2007,41(8):783-788.
APA	Guo, JM.,Yao, F.,Lou, YL.,Xu, CF.,Xiao, BX.,...&Liu, Z.(2007).Detecting carcinoma cells in peripheral blood of patients with hepatocellular carcinoma by Immunomagnetic beads and RT-PCR.JOURNAL OF CLINICAL GASTROENTEROLOGY,41(8),783-788.
MLA	Guo, JM,et al."Detecting carcinoma cells in peripheral blood of patients with hepatocellular carcinoma by Immunomagnetic beads and RT-PCR".JOURNAL OF CLINICAL GASTROENTEROLOGY 41.8(2007):783-788.