Cancer Research Unit, PG Department of Botany, Ramakrishna Mission Vivekananda Centenary College, Rahara, Kolkata 700118, India.
Exp Oncol. 2020 Sep;42(3):178-182. doi: 10.32471/exp-oncology.2312-8852.vol-42-no-3.15086.
Pancreatic cancer is an aggressive malignancy with poor prognosis. New options for its treatment including the use of the substances of natural origin are now on the agenda.
An attempt has been taken to examine the effect of ethanolic extract (EE) of edible mushroom Calocybe indica on PANC-1 and MIAPaCa2 cell lines of pancreatic cancer in vitro.
Study of cell morphology, evaluation of apoptosis by DAPI staining under florescence microscope, anti-proliferative study by MTT assay, lactate dehydrogenase assay, pro-apoptotic and anti-apoptotic protein detection by Western blotting were performed to test effects of Calocybe indica EE on the cells of these two lines. Effect on in vitro migration was studied by scratch method.
Both cell lines treated with 250 or 500 µg/mL of EE underwent morphological changes - became more round and shrunken, with membrane blebbing and the reduced cell confluence. The nuclei of treated cells became condensed and fragmented. The percentage of apoptotic cells increased as EE concentration increased from 100 µg/mL to 500 µg/mL. Anti-proliferative effect was recorded at all concentrations within 100-750 µg/mL. The effect EE on lactate dehydrogenase leakage was concentration-dependent in both cell lines. Western blotting showed that caspase-3 and -9, and p53 protein levels were increased and BcL2 protein was decreased after treatment of the cells with EE compared to the control. PANC-1 cell migration was inhibited by 80.12 ± 4.25% after 24 h treatment of the cells with 500 µg/mL EE compared to the control.
EE of Calocybe indica inhibits the growth and induces apoptosis in pancreatic cancer cell lines in vitro.
胰腺癌是一种侵袭性恶性肿瘤，预后不良。目前，治疗胰腺癌的新方法包括使用天然物质。
本研究旨在探讨食用蘑菇 Calocybe indica 的醇提物（EE）对体外胰腺癌细胞系 PANC-1 和 MIAPaCa2 的影响。
通过 DAPI 染色在荧光显微镜下观察细胞形态，通过 MTT 法、乳酸脱氢酶法检测细胞增殖，通过 Western blot 检测促凋亡和抗凋亡蛋白，研究 Calocybe indica EE 对这两种细胞系的作用。通过划痕法研究 EE 对体外迁移的影响。
用 250µg/mL 或 500µg/mL EE 处理的两种细胞系均发生形态变化，细胞变得更圆、皱缩，细胞膜起泡，细胞融合减少。处理后的细胞核出现浓缩和碎裂。随着 EE 浓度从 100µg/mL 增加到 500µg/mL，凋亡细胞的比例增加。在 100-750µg/mL 浓度范围内，均观察到抗增殖作用。EE 对两种细胞系的乳酸脱氢酶漏出的影响呈浓度依赖性。Western blot 显示，与对照组相比，用 EE 处理细胞后，caspase-3 和 -9 以及 p53 蛋白水平升高，BcL2 蛋白水平降低。与对照组相比，用 500µg/mL EE 处理 PANC-1 细胞 24 小时后，细胞迁移抑制率为 80.12%±4.25%。
Calocybe indica EE 抑制胰腺癌细胞系的生长并诱导其凋亡。
