Silencing TUBB3 Expression Destroys the Tegument and Flame Cells of <i>Echinococcus multilocularis</i> Protoscoleces
Alveolar echinococcosis (AE), caused by infection with the larvae of <i>Echinococcus multilocularis</i>, 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 re...
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MDPI AG
2022-09-01
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| Series: | Animals |
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| author | Qiqi Shi Congshan Liu Lele Huo Yi Tao Haobing Zhang |
| author_facet | Qiqi Shi Congshan Liu Lele Huo Yi Tao Haobing Zhang |
| author_sort | Qiqi Shi |
| collection | DOAJ |
| description | Alveolar echinococcosis (AE), caused by infection with the larvae of <i>Echinococcus multilocularis</i>, 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 <i>E. multilocularis</i> 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 <i>E. multilocularis</i> PSCs. Novel drugs targeting this specific beta-tubulin isotype in <i>E. multilocularis</i> are potential methods for disease control and deserve further attention. |
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| spelling | doaj.art-00577cc1fad943f4ad2719cf2d44f1002023-11-23T14:43:28ZengMDPI AGAnimals2076-26152022-09-011218247110.3390/ani12182471Silencing TUBB3 Expression Destroys the Tegument and Flame Cells of <i>Echinococcus multilocularis</i> ProtoscolecesQiqi Shi0Congshan Liu1Lele Huo2Yi Tao3Haobing Zhang4National 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, ChinaNational 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, ChinaNational 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, ChinaNational 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, ChinaNational 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, ChinaAlveolar echinococcosis (AE), caused by infection with the larvae of <i>Echinococcus multilocularis</i>, 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 <i>E. multilocularis</i> 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 <i>E. multilocularis</i> PSCs. Novel drugs targeting this specific beta-tubulin isotype in <i>E. multilocularis</i> are potential methods for disease control and deserve further attention.https://www.mdpi.com/2076-2615/12/18/2471<i>Echinococcus multilocularis</i>RNAimicrotubuleTUBB3tegumentflame cell |
| spellingShingle | Qiqi Shi Congshan Liu Lele Huo Yi Tao Haobing Zhang Silencing TUBB3 Expression Destroys the Tegument and Flame Cells of <i>Echinococcus multilocularis</i> Protoscoleces Animals <i>Echinococcus multilocularis</i> RNAi microtubule TUBB3 tegument flame cell |
| title | Silencing TUBB3 Expression Destroys the Tegument and Flame Cells of <i>Echinococcus multilocularis</i> Protoscoleces |
| title_full | Silencing TUBB3 Expression Destroys the Tegument and Flame Cells of <i>Echinococcus multilocularis</i> Protoscoleces |
| title_fullStr | Silencing TUBB3 Expression Destroys the Tegument and Flame Cells of <i>Echinococcus multilocularis</i> Protoscoleces |
| title_full_unstemmed | Silencing TUBB3 Expression Destroys the Tegument and Flame Cells of <i>Echinococcus multilocularis</i> Protoscoleces |
| title_short | Silencing TUBB3 Expression Destroys the Tegument and Flame Cells of <i>Echinococcus multilocularis</i> Protoscoleces |
| title_sort | silencing tubb3 expression destroys the tegument and flame cells of i echinococcus multilocularis i protoscoleces |
| topic | <i>Echinococcus multilocularis</i> RNAi microtubule TUBB3 tegument flame cell |
| url | https://www.mdpi.com/2076-2615/12/18/2471 |
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