Manibalan Subramaniyan, Shobana Ayyachamy, Kiruthika Manickam, Achary Anant, Swathi Madasamy, Venkatalakshmi Renganathan, Thirukumaran Kandasamy, Suhasini K, Roopathy Sharon
Centre for Research, Kamaraj College of Engineering and Technology, Madurai, India.
Department of Biotechnology, Kamaraj College of Engineering and Technology, Madurai, India.
Front Bioeng Biotechnol. 2020 May 13;8:328. doi: 10.3389/fbioe.2020.00328. eCollection 2020.
The prevalence of polycystic ovary syndrome (PCOS) has been gradually increasing among adult females worldwide. Laparoscopy drilling on ovary is the only available temporary solution with a high incidence of reoccurrence. S100A8 with S100A9 complex is believed to facilitate the cyst migration in PCOS condition. The high evident protein interaction network studies between PCOS biomarkers, cancer invasion markers, and the interactors of S100A8 confirm that this protein has strong interaction with other selective PCOS biomarkers, which may be associative in the immature cyst invasion process. Through the network studies, intensive structural and pathway analysis, S100A8 is identified as a targetable protein. In this research, the non-SELEX method is adapted to construct RNA Library based on the consensus DNA sequence of Glucocorticoid Response Element (GRE) and screened the best nucleotide fragments which are bound within the active sites of the target protein. Selected sequences are joined as a single strand and screened the one which competitively binds with minimal energy. follow-up of this computational research, the designed RNA aptamer was used to infect the MCF7 cell line through Lipofectamine 2000 mediated delivery to study the anti-cell migration effect. Wound Scratch assay confirms that the synthesized 18-mer oligo has significant inhibition activity toward tumor cell migration at the cellular level.
多囊卵巢综合征（PCOS）在全球成年女性中的患病率一直在逐渐上升。卵巢腹腔镜打孔术是唯一可用的临时解决方案，但复发率很高。S100A8与S100A9复合物被认为在PCOS情况下促进囊肿迁移。PCOS生物标志物、癌症侵袭标志物与S100A8相互作用分子之间的高可信度蛋白质相互作用网络研究证实，该蛋白与其他选择性PCOS生物标志物有很强的相互作用，这可能在未成熟囊肿侵袭过程中存在关联。通过网络研究、深入的结构和通路分析，S100A8被确定为一个可靶向的蛋白质。在本研究中，采用非SELEX方法基于糖皮质激素反应元件（GRE）的共有DNA序列构建RNA文库，并筛选出与靶蛋白活性位点结合的最佳核苷酸片段。将选定的序列连接成单链，并筛选出以最小能量竞争性结合的序列。在这项计算研究的后续实验中，设计的RNA适体通过脂质体2000介导的递送感染MCF7细胞系，以研究其抗细胞迁移作用。伤口划痕试验证实，合成的18聚体寡核苷酸在细胞水平上对肿瘤细胞迁移具有显著的抑制活性。
