College of Biological Science and Engineering, Hebei University of Science and Technology, Shijiazhuang, China.
Laboratory of Veterinary Drug Residues, Hebei Institute of Veterinary Drugs Control, Shijiazhuang, Hebei provience, PR China.
Nutr Cancer. 2022;74(2):640-649. doi: 10.1080/01635581.2021.1897632. Epub 2021 Mar 15.
Lentinan can lead to apoptosis of tumor cells and improve immune function. However, limited research focused on the immunogenic death regulation mechanism of lentinan on mouse H22 cells. The study aimed to explore the effect of Lentinan on the expression of immunogenic death-related proteins in mice H22 cells. MTT method was used to detect and evaluate the effect of 200-1000 μg/mL lentinan on the survival rate of H22 cells after 24 h, 48 h, and 72 h, respectively. Flow cytometry was employed to collect the apoptotic rate of lentinan at different concentrations (200-800μg/mL) on H22 cells for 48 h, and obtain the apoptotic rate of 600 μg/mL lentinan at different times (12-72 h). The effect of Lentinan on the expression of H22 Immunogenic Cell Death proteins was analyzed by ELISA and HPLC-MS afterward. Results suggest that lentinan cytotoxic and pro-apoptotic have a concentration-dependent manner with the H22 cells. Moreover, the rate of apoptosis increased significantly ( < 0.05) in 24 h. Lentinan can induce the expression of Calreticulin(CRT), High mobility protein 1(HMGB1), ATP and Heat shock protein 70 (HSP70) .Therefore, the antitumor effect of lentinan may be related to the regulation of immunogenic death-related protein expression, which was beneficial to the future development of liver cancer vaccines.
香菇多糖可诱导肿瘤细胞凋亡，提高免疫功能。但目前关于香菇多糖诱导小鼠 H22 肝癌细胞免疫原性细胞死亡的调控机制的研究较少。本研究旨在探讨香菇多糖对小鼠 H22 肝癌细胞免疫原性细胞死亡相关蛋白表达的影响。MTT 法检测和评价 200-1000μg/ml 香菇多糖对 H22 细胞 24、48、72 h 存活率的影响。流式细胞术收集不同浓度（200-800μg/ml）香菇多糖作用 48 h 后 H22 细胞的凋亡率，并获得 600μg/ml 香菇多糖作用不同时间（12-72 h）的凋亡率。ELISA 和 HPLC-MS 分析香菇多糖对 H22 免疫原性细胞死亡蛋白表达的影响。结果表明，香菇多糖对 H22 细胞具有浓度依赖性的细胞毒性和促凋亡作用，且在 24 h 时凋亡率显著增加（<0.05）。香菇多糖能诱导钙网织蛋白（CRT）、高迁移率族蛋白 1（HMGB1）、ATP 和热休克蛋白 70（HSP70）的表达。因此，香菇多糖的抗肿瘤作用可能与免疫原性细胞死亡相关蛋白表达的调节有关，这有助于肝癌疫苗的未来发展。
