Subtractive Cell-SELEX Selection of DNA Aptamers Binding Specifically and Selectively to Hepatocellular Carcinoma Cells with High Metastatic Potential.
Relapse and metastasis are two key risk factors of hepatocellular carcinoma (HCC) prognosis; thus, it is emergent to develop an early and accurate detection method for prognostic evaluation of HCC after surgery. In this study, we sought to acquire oligonucleotide DNA aptamers that specifically bind to HCC cells with high metastatic potential. Two HCC cell lines derived from the same genetic background but with different metastatic potential were employed: MHCC97L (low metastatic properties) as subtractive targets and HCCLM9 (high metastatic properties) as screening targets. To mimic a fluid combining environment, initial DNA aptamers library was firstly labelled with magnetic nanoparticles using biotin-streptavidin system and then applied for aptamers selection. Through 10-round selection with subtractive Cell-SELEX, six aptamers, LY-1, LY-13, LY-46, LY-32, LY-27/45, and LY-7/43, display high affinity to HCCLM9 cells and do not bind to MHCC97L cells, as well as other tumor cell lines, including breast cancer, lung cancer, colon adenocarcinoma, gastric cancer, and cervical cancer, suggesting high specificity for HCCLM9 cells. Thus, the aptamers generated here will provide solid basis for identifying new diagnostic targets to detect HCC metastasis and also may provide valuable clues for developing new targeted therapeutics.
Department of Laboratory Medicine and Center for Gene Diagnosis, Zhongnan Hospital of Wuhan University, 169 Donghu Road, Wuchang District, Wuhan 430071, China.
Hao
H
Chen
Department of Immunology, School of Basic Medical Sciences, Wuhan University, 185 Donghu Road, Wuchang District, Wuhan 430071, China.
Chun-Hui
CH
Yuan
Department of Immunology, School of Basic Medical Sciences, Wuhan University, 185 Donghu Road, Wuchang District, Wuhan 430071, China.
Yi-Fei
YF
Yang
Department of Laboratory Medicine and Center for Gene Diagnosis, Zhongnan Hospital of Wuhan University, 169 Donghu Road, Wuchang District, Wuhan 430071, China.
Chang-Qing
CQ
Yin
Department of Immunology, School of Basic Medical Sciences, Wuhan University, 185 Donghu Road, Wuchang District, Wuhan 430071, China.
Qing
Q
Guan
Department of Laboratory Medicine and Center for Gene Diagnosis, Zhongnan Hospital of Wuhan University, 169 Donghu Road, Wuchang District, Wuhan 430071, China.
Fu-Bing
FB
Wang
Department of Laboratory Medicine and Center for Gene Diagnosis, Zhongnan Hospital of Wuhan University, 169 Donghu Road, Wuchang District, Wuhan 430071, China.
Jian-Cheng
JC
Tu
D052157:Aptamers, Nucleotide / Q000627:therapeutic use*; D001665:Binding Sites; D001710:Biotin; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology* / Q000628:therapy; D045744:Cell Line, Tumor; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology* / Q000628:therapy; D009362:Neoplasm Metastasis; D009364:Neoplasm Recurrence, Local; D052156:SELEX Aptamer Technique; D019809:Streptavidin
D052157:Aptamers, Nucleotide; D001710:Biotin; D019809:Streptavidin