Department of Genetics & Biotechnology, Osmania University, Hyderabad, Telangana, 500 007, India.
School of Biosciences Engineering and Technology, VIT Bhopal University, Bhopal, Madhya Pradesh, 466114, India.
Med Oncol. 2022 Sep 1;39(11):179. doi: 10.1007/s12032-022-01826-5.
The DEAD-box helicase family member DDX3 is involved in many diseases, such as viral infection, inflammation, and cancer. Many studies in the last decade have revealed the role of DDX3 in tumorigenesis and metastasis. DDX3 has both tumour suppressor and oncogenic effect, in the present study we have evaluated the expression levels of DDX3 in cervical squamous cell carcinoma at mRNA level via real-time PCR and protein level via Immunohistochemistry. DDX3 has become a molecule of interest in cancer biology that promotes drug resistance by adaptive response inevitably leading to treatment failure. One approach to avoid the development of resistant to disease is to create novel drugs that target the overexpressed proteins, we designed and synthesized a novel 7-azaindole derivative (7-AID) compound, {5-[1H-pyrrolo (2, 3-b) pyridin-5-yl] pyridin-2-ol]} that could lodge within the adenosine-binding pocket of the DDX3 (PDB ID: 2I4I). The binding efficacy of 7-AID compound with DDX3 was analysed by molecular docking studies. 7-AID was found to interact with the key residues Tyr200 and Arg202 from the Q-motif rendered by π-interactions and hydrogen bonds within the binding pocket with good docking score - 7.99 kcal/mol. The cytotoxicity effect of 7-AID compound was evaluated using MTT assay on human cervical carcinoma cells (HeLa) and breast cancer cells (MCF-7 and MDA MB-231) and the compound shown effective inhibitory concentration (IC) on Hela cells 16.96 µM/ml and 14.12 and 12.69 µM/ml on MCF-7 and MDA MB-231, respectively. Further, the in-vitro, in-vivo anti-cancer and anti-angiogenic assessment of 7-AID compound was evaluated on Hela cells using scratch wound-healing assay, DAPI staining, cell cycle analysis, immunoblotting, and chorioallontoic membrane assay. Furthermore, the inhibitory effect of derivative compound on DDX3 was investigated in HeLa, MCF-7, and MDA MB-231 cells at the mRNA and protein levels. The results showed that the 7-AID compound effectively inhibited DDX3 in a dose-dependent manner, and the findings suggest that the compound could be used as a potential DDX3 inhibitor.
DEAD 盒解旋酶家族成员 DDX3 参与多种疾病，如病毒感染、炎症和癌症。过去十年的许多研究揭示了 DDX3 在肿瘤发生和转移中的作用。DDX3 具有肿瘤抑制和致癌作用，本研究通过实时 PCR 评估了宫颈鳞状细胞癌中 DDX3 的 mRNA 水平，通过免疫组织化学评估了 DDX3 的蛋白水平。DDX3 已成为癌症生物学中一个有研究意义的分子，通过适应性反应促进耐药性，不可避免地导致治疗失败。避免疾病耐药性发展的一种方法是设计针对过度表达蛋白的新型药物，我们设计并合成了一种新型的 7-氮杂吲哚衍生物（7-AID）化合物，{5-[1H-吡咯并[2,3-b]吡啶-5-基]吡啶-2-醇}，该化合物可以位于 DDX3 的腺苷结合口袋中（PDB ID：2I4I）。通过分子对接研究分析了 7-AID 化合物与 DDX3 的结合效力。发现 7-AID 与 Q 基序中的关键残基 Tyr200 和 Arg202 相互作用，通过 π 相互作用和氢键在结合口袋中具有良好的对接评分-7.99 kcal/mol。通过 MTT 测定法评估了 7-AID 化合物对人宫颈癌细胞（HeLa）和乳腺癌细胞（MCF-7 和 MDA MB-231）的细胞毒性作用，该化合物对 Hela 细胞的有效抑制浓度（IC）为 16.96 μM/ml，对 MCF-7 和 MDA MB-231 的 IC 分别为 14.12 和 12.69 μM/ml。此外，还通过划痕愈合试验、DAPI 染色、细胞周期分析、免疫印迹和绒毛尿囊膜试验在 Hela 细胞上评估了 7-AID 化合物的体内、体内抗癌和抗血管生成作用。此外，还在 HeLa、MCF-7 和 MDA MB-231 细胞中研究了衍生物化合物对 DDX3 的抑制作用。结果表明，7-AID 化合物以剂量依赖性方式有效抑制 DDX3，研究结果表明，该化合物可用作潜在的 DDX3 抑制剂。
