Perez-Matas Edgar, Hanano Abdulsamie, Moyano Elisabeth, Bonfill Mercedes, Cusido Rosa M, Palazon Javier
Secció de Fisiologia Vegetal, Facultat de Farmacia i Ciències de l'Alimentació, Universitat de Barcelona, Barcelona, Spain.
Department of Molecular Biology and Biotechnology, Atomic Energy Commission of Syria, Damascus, Syria.
Front Plant Sci. 2022 Jul 29;13:942433. doi: 10.3389/fpls.2022.942433. eCollection 2022.
More knowledge is needed about the molecular/cellular control of paclitaxel (PTX) production in spp. cell cultures. In this study, the yield of this anticancer agent in cell suspensions was improved 11-fold after elicitation with coronatine (COR) compared to the untreated cells, and 18-fold when co-supplemented with methyl-β-cyclodextrins (β-CDs). In the dual treatment, the release of taxanes from the producer cells was greatly enhanced, with 81.6% of the total taxane content being found in the medium at the end of the experiment. The experimental conditions that caused the highest PTX production also induced its maximum excretion, and increased the expression of taxane biosynthetic genes, especially the flux-limiting BAPT and DBTNBT. The application of COR, which activates PTX biosynthesis, together with β - CDs, which form inclusion complexes with PTX and related taxanes, is evidently an efficient strategy for enhancing PTX production and release to the culture medium. Due to the recently described role of lipid droplets (LDs) in the trafficking and accumulation of hydrophobic taxanes in spp. cell cultures, the structure, number and taxane storage capacity of these organelles was also studied. In elicited cultures, the number of LDs increased and they mainly accumulated taxanes with a side chain, especially PTX. Thus, PTX constituted up to 50-70% of the total taxanes found in LDs throughout the experiment in the COR + β - CD-treated cultures. These results confirm that LDs can store taxanes and distribute them inside and outside cells.
关于红豆杉属植物细胞培养中紫杉醇（PTX）产生的分子/细胞控制，我们还需要更多的知识。在本研究中，与未处理的细胞相比，用冠菌素（COR）诱导后，这种抗癌药物在红豆杉细胞悬浮液中的产量提高了11倍，而当与甲基-β-环糊精（β-CDs）共同添加时，产量提高了18倍。在双重处理中，紫杉烷从生产细胞中的释放大大增强，在实验结束时，培养基中发现了总紫杉烷含量的81.6%。导致最高PTX产量的实验条件也诱导了其最大排泄，并增加了紫杉烷生物合成基因的表达，尤其是通量限制的BAPT和DBTNBT。激活PTX生物合成的COR与能与PTX及相关紫杉烷形成包合物的β - CDs一起应用，显然是提高PTX产量并释放到培养基中的有效策略。由于最近描述了脂滴（LDs）在红豆杉属植物细胞培养中疏水性紫杉烷的运输和积累中的作用，因此还研究了这些细胞器的结构、数量和紫杉烷储存能力。在诱导培养中，LDs的数量增加，并且它们主要积累带有侧链的紫杉烷，尤其是PTX。因此，在COR + β - CD处理的培养物中，在整个实验过程中，PTX占LDs中发现的总紫杉烷的50 - 70%。这些结果证实LDs可以储存紫杉烷并在细胞内外进行分配。
