Shi Qiqi, Liu Congshan, Huo Lele, Tao Yi, Zhang Haobing
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention (Chinese Center for Tropical Diseases Research), NHC Key Laboratory of Parasite and Vector Biology, WHO Collaborating Centre for Tropical Diseases, National Center for International Research on Tropical Diseases, Shanghai 200025, China.
Animals (Basel). 2022 Sep 19;12(18):2471. doi: 10.3390/ani12182471.
Alveolar echinococcosis (AE), caused by infection with the larvae of , is a neglected tropical disease and zoonosis that causes remarkable morbidity in humans and has economic importance in the livestock industry worldwide. The growth of this parasite resembles the invasion and proliferation of malignant tumours. Microtubules, especially the β-tubulin subunit in the exposed end, are the targets of many antitumour drugs. However, the role of TUBB3, which is the most studied isotype in solid tumours and is also a marker of biological aggressiveness associated with the modulation of tumour metastatic abilities in the growth and development of platyhelminths, is unknown. In this study, protoscoleces (PSCs) are cultivated in monophasic medium in vitro. Using electroporated short interfering RNA (siRNA), EmTUBB3 knockdown was performed with two EmTUBB3-specific siRNAs (siRNA-1 and siRNA-2). qRT-PCR was performed to detect the expression of TUBB3. PSCs viability and the evagination rate and number of body contractions were quantified under a light microscope. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to observe the ultra-morphological changes of the parasites. After siRNA interference, the EmTUBB3 expression in PSCs was significantly reduced. Reduced viability, a decreased evagination rate and a decreased number of body contractions were also documented. In particular, shrinkage and roughness of the tegument were observed. Ultrastructural changes included marked damage to flame cells, cracked cilia structures enclosed in the cell body and ruptured microtubule structures. EmTUBB3 possibly plays a crucial role in tegument and flame cell integrity in PSCs. Novel drugs targeting this specific beta-tubulin isotype in are potential methods for disease control and deserve further attention.
泡型包虫病（AE）由 的幼虫感染引起，是一种被忽视的热带疾病和人畜共患病，可导致人类显著发病，并在全球畜牧业中具有经济重要性。这种寄生虫的生长类似于恶性肿瘤的侵袭和增殖。微管，尤其是暴露端的β-微管蛋白亚基，是许多抗肿瘤药物的靶点。然而，TUBB3的作用尚不清楚，TUBB3是实体瘤中研究最多的亚型，也是与扁形动物生长发育过程中肿瘤转移能力调节相关的生物学侵袭性标志物。在本研究中，原头节（PSCs）在单相培养基中进行体外培养。使用电穿孔短干扰RNA（siRNA），用两种EmTUBB3特异性siRNA（siRNA-1和siRNA-2）进行EmTUBB3基因敲低。进行qRT-PCR检测TUBB3的表达。在光学显微镜下对PSCs的活力、外翻率和体收缩次数进行量化。使用扫描电子显微镜（SEM）和透射电子显微镜（TEM）观察寄生虫的超微形态变化。siRNA干扰后，PSCs中EmTUBB3的表达显著降低。还记录到活力降低、外翻率降低和体收缩次数减少。特别是，观察到皮层收缩和粗糙。超微结构变化包括焰细胞明显受损、包裹在细胞体内的纤毛结构破裂和微管结构断裂。EmTUBB3可能在PSCs的皮层和焰细胞完整性中起关键作用。针对该特定β-微管蛋白亚型的新型药物是疾病控制的潜在方法，值得进一步关注。
