Zhang Qiang, Wang Guihua, Xie Yongfang, Gao Zhiqin, Liang Zumu, Pan Zhifang, Wang Guohui, Feng Weiguo
College of Bioscience and Technology, Weifang Medical University, Weifang, China.
Department of Fundamental Veterinary, College of Veterinary Medicine, Shandong Agricultural University, Tai'an, China.
Front Pharmacol. 2020 Feb 26;11:89. doi: 10.3389/fphar.2020.00089. eCollection 2020.
Microcystin-leucine arginine (MC-LR) is a potent tumor initiator that can induce malignant cell transformation. Cellular mechanical characteristics are pivotal parameters that are closely related to cell invasion. The aim of this study is to determine the effect of MC-LR on mechanical parameters, microfilament, and cell invasion in DU145 and WPMY cells. Firstly, 10 μM MC-LR was selected as the appropriate concentration cell viability assay. Subsequently, after MC-LR treatment, the cellular deformability and viscoelastic parameters were tested using the micropipette aspiration technique. The results showed that MC-LR increased the cellular deformability, reduced the cellular viscoelastic parameter values, and caused the cells to become softer. Furthermore, microfilament and microfilament-associated proteins were examined by immunofluorescence and Western blot, respectively. Our results showed that MC-LR induced microfilament reorganization and increased the expression of p-VASP and p-ezrin. Finally, the impact of MC-LR on cell invasion was evaluated. The results revealed that MC-LR promoted cell invasion. Taken together, our results suggested that mechanical changes and microfilament reorganization were involved in MC-LR-promoted cell invasion in DU145 and WPMY cells. Our data provide novel information to explain the toxicological mechanism of MC-LR.
微囊藻毒素 - 亮氨酸精氨酸（MC-LR）是一种强效的肿瘤起始剂，可诱导恶性细胞转化。细胞力学特性是与细胞侵袭密切相关的关键参数。本研究的目的是确定MC-LR对DU145和WPMY细胞的力学参数、微丝和细胞侵袭的影响。首先，选择10 μM MC-LR作为合适的浓度进行细胞活力测定。随后，在MC-LR处理后，使用微量移液器吸液技术测试细胞的变形能力和粘弹性参数。结果表明，MC-LR增加了细胞的变形能力，降低了细胞的粘弹性参数值，并使细胞变得更柔软。此外，分别通过免疫荧光和蛋白质印迹法检测微丝和微丝相关蛋白。我们的结果表明，MC-LR诱导微丝重组并增加了p-VASP和p-埃兹蛋白的表达。最后，评估了MC-LR对细胞侵袭的影响。结果显示，MC-LR促进了细胞侵袭。综上所述，我们的结果表明，力学变化和微丝重组参与了MC-LR促进DU145和WPMY细胞侵袭的过程。我们的数据为解释MC-LR的毒理学机制提供了新的信息。
