Identification of an aptamer through whole cell-SELEX for targeting high metastatic liver cancers
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Yuan Rong1, Hao Chen1, Xue-Feng Zhou2, Chang-Qing Yin1, Bi-Cheng Wang3, Chun-Wei Peng4, Shao-Ping Liu5, Fu-Bing Wang1
1Department of Laboratory Medicine & Center for Gene Diagnosis, Zhongnan Hospital of Wuhan University, Wuchang, Wuhan 430071, P.R. China
2Department of Thoracic and Cardiovascular Surgery, Zhongnan Hospital of Wuhan University, Wuchang, Wuhan 430071, P.R. China
3Department of Pathology, Zhongnan Hospital of Wuhan University, Wuchang, Wuhan 430071, P.R. China
4Department of Oncology, Zhongnan Hospital of Wuhan University, Wuchang, Wuhan 430071, P.R. China
5Hubei Key Laboratory of Tumor Biological Behaviors & Hubei Cancer Clinical Study Center, Wuchang, Wuhan 430071, P.R. China
Fu-Bing Wang, e-mail: firstname.lastname@example.org
Keywords: hepatocellular carcinoma, migration, invasion, cell-SELEX, aptamer
Received: May 12, 2015 Accepted: January 13, 2016 Published: January 23, 2016
Hepatocellular carcinoma (HCC) is one of the most deadly human cancers due to its ability of invasion and metastasis. Thus, the approaches to identify potential compounds that inhibit invasion and metastasis of HCC are critical for treatment of this disease. In the present study, we used HCCLM9 cells with high metastatic potential and MHCC97L with low metastatic potential as a model system to study the molecular mechanisms of HCC metastasis. By applying cell- Systematic Evolution of Ligands by Exponential enrichment (SELEX) against living cells, we used HCCLM9 as target cells and MHCC97L cells as control to screen a group of HCC metastasis- and cell-specific DNA aptamers. One of selected aptamers, LY-1, could specifically bind to metastatic HCC with a dissociation constant (Kd) in nanomolar range. In vitro studies demonstrated that LY-1 can recognize and bind to membrane protein of metastatic HCC cells. Furthermore, QD605 labeled LY-1 aptamer could recognize HCC cells in both local liver cancer tissues and pulmonary metastatic sites in a xenograft model of HCC with pulmonary metastasis. Further biochemical and immunostaining studies showed that LY-1 could selectively bind to a subpopulation of more metastatic cells in HCCLM9 cells, which express more CK19 and vimentin. Finally, treatment of highly metastatic cells with LY-1 led to reduced migration and invasiveness of HCCLM9 cells in vitro and suppression of xenograft growth in vivo. Taken together, the present study demonstrated the tumor targeting and tumor suppressive effects of LY-1, which could be a promising molecular probe for metastatic HCC and a potential candidate of chemotherapy for metastatic HCC.
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