Identification of an aptamer through whole cell-SELEX for targeting high metastatic liver cancers.
Identification of an aptamer through whole cell-SELEX for targeting high metastatic liver cancers.
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DOI：
10.18632/oncotarget.6988
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发表时间：
2016-02-16
期刊：
影响因子：
--
通讯作者：
Wang FB
中科院分区：
文献类型：
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作者：
Rong Y;Chen H;Zhou XF;Yin CQ;Wang BC;Peng CW;Liu SP;Wang FB
肝细胞癌（HCC）因其侵袭和转移能力而成为最致命的人类癌症之一。因此，鉴定抑制 HCC 侵袭和转移的潜在化合物的方法对于治疗该疾病至关重要。在本研究中，我们使用高转移潜能的HCCLM9细胞和低转移潜能的MHCC97L细胞作为模型系统来研究HCC转移的分子机制。通过对活细胞应用细胞配体指数富集系统进化（SELEX），我们以 HCCLM9 为靶细胞，MHCC97L 细胞为对照，筛选出一组 HCC 转移和细胞特异性 DNA 适体。选定的适体之一 LY-1 可以与转移性 HCC 特异性结合，解离常数 (Kd) 在纳摩尔范围内。体外研究表明，LY-1 可以识别并结合转移性 HCC 细胞的膜蛋白。此外，QD605标记的LY-1适体可以识别局部肝癌组织和肺转移HCC异种移植模型中肺转移部位的HCC细胞。进一步的生化和免疫染色研究表明，LY-1 可以选择性地与 HCCLM9 细胞中更多转移细胞的亚群结合，这些细胞表达更多的 CK19 和波形蛋白。最后，用 LY-1 处理高度转移的细胞可减少 HCCLM9 细胞的体外迁移和侵袭性，并抑制体内异种移植物的生长。综上所述，本研究证明了 LY-1 的肿瘤靶向和肿瘤抑制作用，其可能成为转移性 HCC 的有前途的分子探针和转移性 HCC 化疗的潜在候选者。
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.