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白头翁皂苷 E 通过负向调控脂筏中的 flotillin-2 抑制 Akt/FASN 通路抑制非小细胞肺癌细胞的活力、迁移、侵袭并促进其凋亡。

Pulsatilla saponin E suppresses viability, migration, invasion and promotes apoptosis of NSCLC cells through negatively regulating Akt/FASN pathway inhibition of flotillin-2 in lipid raft.

机构信息

Department of Cardiothoracic Surgery, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, PR China.

Department of Cardiovascular Medicine, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, PR China.

出版信息

J Recept Signal Transduct Res. 2022 Feb;42(1):23-33. doi: 10.1080/10799893.2020.1839764. Epub 2020 Nov 26.

Abstract

PURPOSE

Pulsatilla saponins from have potential anti-tumor activities to certain human cancers. However, the roles of pulsatilla saponin E separated from pulsatilla saponins in non-small cell lung cancer (NSCLC) have not been reported.

MATERIALS AND METHODS

After treating NSCLC cells by pulsatilla saponin E at different concentrations, cell viability was measured by MTT and CCK-8 assays, and cell migration, invasion and apoptosis were detected by scratch wound-healing, transwell and flow cytometry assays. The contents of free cholesterol (FC) and total cholesterol (TC) were measured by high performance liquid chromatography (HPLC). The expression levels of flotillin-1, flotillin-2, Akt, fatty acid synthase (FASN) were detected by qRT-PCR and Western blot assays.

RESULTS

Pulsatilla saponin E suppressed viability, migration, invasion and promoted apoptosis of NSCLC cells followed by regulation of apoptosis-related proteins, reduced contents of FC and TC, and the expression levels of flotillin-1, flotillin-2, Akt, and FASN in a concentration-dependent manner. However, the inhibitory effects of pulsatilla saponin E on viability, migration, invasion of A549 cells and the expression levels of flotillin-1, flotillin-2, Akt, and FASN were reversed by flotillin-2 overexpression.

CONCLUSIONS

Our study revealed that pulsatilla saponin E suppressed migration, invasion and promoted apoptosis of NSCLC cells through negatively regulating Akt/FASN signaling pathway the inhibition of flotillin-2 in lipid raft (LR). The current findings could be explored for developing a novel therapeutic drug for NSCLC treatment.

摘要

目的

白头翁皂苷来自白头翁具有潜在的抗肿瘤活性对某些人类癌症。然而,白头翁皂苷 E 从白头翁皂苷分离在非小细胞肺癌(NSCLC)中的作用尚未报道。

材料与方法

用不同浓度的白头翁皂苷 E 处理 NSCLC 细胞后,用 MTT 和 CCK-8 法测定细胞活力,划痕愈合、transwell 和流式细胞术检测细胞迁移、侵袭和凋亡。用高效液相色谱法(HPLC)测定游离胆固醇(FC)和总胆固醇(TC)的含量。用 qRT-PCR 和 Western blot 法检测 flotillin-1、flotillin-2、Akt、脂肪酸合酶(FASN)的表达水平。

结果

白头翁皂苷 E 抑制 NSCLC 细胞活力、迁移、侵袭,促进细胞凋亡,同时调节凋亡相关蛋白,降低 FC 和 TC 含量,浓度依赖性地上调 flotillin-1、flotillin-2、Akt 和 FASN 的表达水平。然而,白头翁皂苷 E 对 A549 细胞活力、迁移、侵袭的抑制作用以及 flotillin-1、flotillin-2、Akt 和 FASN 的表达水平被 flotillin-2 过表达逆转。

结论

本研究表明,白头翁皂苷 E 通过负调控 Akt/FASN 信号通路抑制脂质筏(LR)中的 flotillin-2,抑制 NSCLC 细胞的迁移、侵袭并促进细胞凋亡。这一发现为开发 NSCLC 治疗的新型治疗药物提供了依据。

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