Department of Histology and Embryology, Faculty of Medicine and Dentistry, Palacky University, 779 00 Olomouc, Czech Republic.
Department of Histology and Embryology, Faculty of Medicine and Dentistry, Palacky University, 779 00 Olomouc, Czech Republic.
Biochim Biophys Acta Gen Subj. 2023 Dec;1867(12):130496. doi: 10.1016/j.bbagen.2023.130496. Epub 2023 Oct 20.
Intestinal epithelial differentiation is a highly organised process. It is influenced by a variety of signalling pathways and enzymes, such as the PI3K pathway and soluble epoxide hydrolase (sEH) from arachidonic acid metabolism. We investigated the changes in the expression of enzymes and lipid messenger from the PI3K pathway, including PTEN, during intestinal cell differentiation in vitro using HT-29 and Caco2 cells and compared them with immunohistochemical patterns of these proteins in human colon. To investigate the possible crosstalk between the PI3K pathway and sEH, we treated HT-29 and Caco2 cells with the sEH inhibitor TPPU. Administration of TPPU to differentiated cells decreased the expression of PTEN, thus reversing the change in its expression observed during cell differentiation. In addition, multiplex immunofluorescence staining confirmed the relationship between the expression of PTEN and villin, a marker of intestinal cell differentiation, ranging from a moderate correlation in undifferentiated cells to a very strong correlation in differentiated cells treated with TPPU. Furthermore, we confirm that PTEN and sEH mirrored their expression patterns in samples of prenatal and adult human intestine compared to tumours using immunohistochemical staining. Taken together, it appears that PTEN and sEH cooperate in the process of intestinal cell differentiation. A better understanding of the crosstalk between the PI3K pathway and sEH and its consequences for cell differentiation is highly desirable, as several sEH inhibitors are under clinical investigation for the treatment of various diseases.
肠上皮细胞分化是一个高度有序的过程。它受到多种信号通路和酶的影响，如 PI3K 通路和来自花生四烯酸代谢的可溶性环氧化物水解酶 (sEH)。我们使用 HT-29 和 Caco2 细胞在体外研究了 PI3K 通路中包括 PTEN 在内的酶和脂质信使在肠细胞分化过程中的表达变化，并将其与这些蛋白质在人结肠中的免疫组织化学模式进行了比较。为了研究 PI3K 通路和 sEH 之间可能的串扰，我们用 sEH 抑制剂 TPPU 处理 HT-29 和 Caco2 细胞。向分化细胞中给予 TPPU 会降低 PTEN 的表达，从而逆转细胞分化过程中观察到的其表达变化。此外，多重免疫荧光染色证实了 PTEN 和微管蛋白之间的表达关系，微管蛋白是肠细胞分化的标志物，在未分化细胞中呈中度相关，在用 TPPU 处理的分化细胞中呈很强的相关性。此外，我们通过免疫组织化学染色证实，与肿瘤相比，PTEN 和 sEH 在产前和成人肠道样本中的表达模式与其在肿瘤中的表达模式相匹配。综上所述，PTEN 和 sEH 似乎在肠细胞分化过程中合作。更好地了解 PI3K 通路和 sEH 之间的串扰及其对细胞分化的影响是非常可取的，因为几种 sEH 抑制剂正在临床研究中用于治疗各种疾病。
