Gandomkari Maryam Shahbazi, Ayat Hoda, Ahadi Ali Mohammad
Department of Genetics, Shahrekord University, Rahbar Blvd, P O Box 115, 8818634141, Shahrekord, IR, Iran.
Biotechnol Lett. 2022 Jun;44(5-6):703-712. doi: 10.1007/s10529-022-03254-x. Epub 2022 May 7.
Gliomas are highly invasive and lethal malignancy that do not respond to current therapeutic approaches. Novel therapeutic agents are required to target molecular mechanisms involved in glioma progression. MeICT is a new short-chain toxin isolated from Mesobuthus eupeus scorpion venom. This toxin contained 34 amino acid residues and belongs to chloride channels toxins. In this study, the coding sequence of MeICT was cloned into the pET32Rh vector and a high yield of soluble recombinant MeICT was expressed and purified. Recombinant MeICT-His significantly inhibited the proliferation and migration of glioma cells at low concentration. In vivo studies showed that MeICT was not toxic when administrated to mice at high doses. We also determined the effect of MeICT on the mRNA expression of MMP-2, Annexin A2 and FOXM-2 that are key molecules in the progression and invasion of glioma. Expression of Annexin A2 and FOXM1 mRNA was significantly down-regulated following treatment with MeICT. However, no significant decrease in the expression of MMP-2 gene was identified. In this study a short toxin with four disulfide bonds was successfully produced and its anti-cancer effects was detected. Our findings suggest that recombinant MeICT can be considered as a new potent agent for glioma targeting.
神经胶质瘤是一种具有高度侵袭性和致命性的恶性肿瘤,对目前的治疗方法没有反应。需要新型治疗药物来靶向参与神经胶质瘤进展的分子机制。MeICT是一种从东亚钳蝎毒液中分离出的新型短链毒素。这种毒素含有34个氨基酸残基,属于氯离子通道毒素。在本研究中,将MeICT的编码序列克隆到pET32Rh载体中,表达并纯化了高产量的可溶性重组MeICT。重组MeICT-His在低浓度下显著抑制神经胶质瘤细胞的增殖和迁移。体内研究表明,高剂量给予小鼠MeICT时无毒。我们还确定了MeICT对MMP-2、膜联蛋白A2和FOXM-2 mRNA表达的影响,这些是神经胶质瘤进展和侵袭中的关键分子。用MeICT处理后,膜联蛋白A2和FOXM1 mRNA的表达显著下调。然而,未发现MMP-2基因表达有显著降低。在本研究中,成功制备了一种具有四个二硫键的短毒素,并检测了其抗癌作用。我们的研究结果表明,重组MeICT可被视为一种新的有效的神经胶质瘤靶向药物。