Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science and Technology (Deemed to be University), Chennai, Tamilnadu, India.
School of Health Sciences, Purdue University, 550 Stadium Mall Drive, West Lafayette, IN, 47907, USA.
Tissue Eng Regen Med. 2024 Feb;21(2):353-366. doi: 10.1007/s13770-023-00599-w. Epub 2023 Nov 7.
Regeneration is a highly complex process that requires the coordination of numerous molecular events, and identifying the key ruler that governs is important to investigate. While it has been shown that TCTP is a multi-functional protein that regulates cell proliferation, differentiation, apoptosis, anti-apoptosis, stem cell maintenance, and immune responses, but only a few studies associated to regeneration have been reported. To investigate the multi-functional role of TCTP in regeneration, the earthworm Perionyx excavatus was chosen.
Through pharmacological suppression of TCTP, amputation, histology, molecular docking, and western blotting, the multi-function role of TCTP involved in regeneration is revealed.
Amputational studies show that P. excavatus is a clitellum-independent regenerating earthworm resulting in two functional worms upon amputation. Arresting cell cycle at the G1/S boundary using 2 mM Thymidine confirms that P. excavatus execute both epimorphosis and morphallaxis regeneration mode. The pharmacological suppression of TCTP using buclizine results in regeneration suppression. Following the combinatorial injection of 2 mM Thymidine and buclizine, the earthworm regeneration is completely blocked, which suggests a critical functional role of TCTP in morphallaxis. The pharmacological inhibition of TCTP also suppresses the key proteins involved in regeneration: Wnt3a (stem cell marker), PCNA (cell proliferation) and YAP1 (Hippo signalling) but augments the expression of cellular stress protein p53.
The collective results indicate that TCTP synchronously is involved in the process of stem cell activation, cell proliferation, morphallaxis, and organ development in the regeneration event.
再生是一个高度复杂的过程，需要协调众多分子事件，鉴定关键调控因子对于研究再生非常重要。虽然已经证明 TCTP 是一种具有多种功能的蛋白质，可调节细胞增殖、分化、凋亡、抗凋亡、干细胞维持和免疫反应，但仅有少数与再生相关的研究被报道。为了研究 TCTP 在再生中的多功能作用，选择了正蚓属（Perionyx）的一种蚓，即环毛蚓（Perionyx excavatus）。
通过 TCTP 的药理学抑制、截断、组织学、分子对接和 Western blot 实验，揭示了 TCTP 在再生中涉及的多功能作用。
截断实验表明，环毛蚓是一种无节间体再生的蚯蚓，在截断后会形成两个具有功能的个体。使用 2mM 胸苷将细胞周期阻滞在 G1/S 边界，证实环毛蚓执行了胚胎型再生和组织型再生两种模式。使用丁布曲明（buclizine）药理学抑制 TCTP 会导致再生受到抑制。联合注射 2mM 胸苷和丁布曲明后，蚯蚓的再生完全被阻断，这表明 TCTP 在组织型再生中具有关键的功能作用。TCTP 的药理学抑制也抑制了参与再生的关键蛋白：Wnt3a（干细胞标志物）、PCNA（细胞增殖）和 YAP1（Hippo 信号通路），但增加了细胞应激蛋白 p53 的表达。
综合结果表明，TCTP 同步参与了再生过程中的干细胞激活、细胞增殖、组织型再生和器官发育过程。
