PMID- 37326754 OWN - NLM STAT- MEDLINE DCOM- 20230731 LR - 20230829 IS - 1573-4978 (Electronic) IS - 0301-4851 (Linking) VI - 50 IP - 8 DP - 2023 Aug TI - Mechanisms of autophagy and endoplasmic reticulum stress in the reversal of platinum resistance of epithelial ovarian cancer cells by naringin. PG - 6457-6468 LID - 10.1007/s11033-023-08558-3 [doi] AB - OBJECTIVE: Our previous studies showed that naringin (Nar) can effectively reverse the cisplatin resistance of ovarian cancer cells. This study aims to explore the potential mechanism by which Nar reverses cisplatin resistance in ovarian cancer. METHODS: The proliferative activity of cells was evaluated using CCK8 and cell clone formation assays. Autophagic flux in cells was evaluated via LC3B immunofluorescence and monodansylcadaverine (MDC) staining. The expression levels of autophagy, endoplasmic reticulum (ER) stress, and apoptosis-related proteins were detected via Western blotting. Autophagy and ER stress were regulated using siATG5, siLC3B, rapamycin (Rap), chloroquine (CQ), 4-phenylbutyric acid (4-PBA), and thapsigargin (TG). siATG5 and siLC3B are short interfering RNAs (siRNAs) used to knock down the expression of ATG5 and LC3B genes, respectively. RESULTS: Nar inhibited autophagy in SKOV3/DDP cells by activating the PI3K/AKT/mTOR pathway. And Nar increased the levels of ER stress-related proteins, namely, P-PERK, GRP78, and CHOP, and promoted apoptosis in SKOV3/DDP cells. Moreover, treatment with the inhibitor of ER stress alleviated apoptosis induced by Nar in SKOV3/DDP cells. In addition, compared to cisplatin or naringin alone, the combination of Nar and cisplatin significantly reduced the proliferative activity of SKOV3/DDP cells. And siATG5, siLC3B, CQ or TG pretreatment further inhibited the proliferative activity of SKOV3/DDP cells. Conversely, Rap or 4-PBA pretreatment alleviated the cell proliferation inhibition caused by Nar combined with cisplatin. CONCLUSION: Nar not only inhibited the autophagy in SKOV3/DDP cells by regulating the PI3K/AKT/mTOR signalling pathway, but also promoted apoptosis in SKOV3/DDP cells by targeting ER stress. Nar can reverse the cisplatin resistance in SKOV3/DDP cells through these two mechanisms. CI - (c) 2023. The Author(s), under exclusive licence to Springer Nature B.V. FAU - Zhu, Jun AU - Zhu J AD - The Third Affiliated Hospital of Nanchang University, The First Hospital of Nanchang City, Nanchang, Jiangxi, China. AD - Jiangxi Provincial Key Laboratory of Tumor Metastasis and Precision Therapy, Nanchang, Jiangxi, China. AD - Nanchang Key Laboratory of Precision Therapy for Gynecological Neoplasms, Nanchang, Jiangxi, China. FAU - Lin, Shixin AU - Lin S AD - The Third Affiliated Hospital of Nanchang University, The First Hospital of Nanchang City, Nanchang, Jiangxi, China. AD - Jiangxi Provincial Key Laboratory of Tumor Metastasis and Precision Therapy, Nanchang, Jiangxi, China. AD - Nanchang Key Laboratory of Precision Therapy for Gynecological Neoplasms, Nanchang, Jiangxi, China. FAU - Zou, Xia AU - Zou X AD - Department of Gynecologic Oncology, Jiangxi Cancer Hospital, Nanchang, Jiangxi, China. FAU - Chen, Xintong AU - Chen X AD - Department of Gynecologic Oncology, Jiangxi Cancer Hospital, Nanchang, Jiangxi, China. FAU - Liu, Yanlan AU - Liu Y AD - The Third Affiliated Hospital of Nanchang University, The First Hospital of Nanchang City, Nanchang, Jiangxi, China. FAU - Yang, Xiaorong AU - Yang X AD - Department of Gynecologic Oncology, Jiangxi Cancer Hospital, Nanchang, Jiangxi, China. FAU - Gao, Jun AU - Gao J AD - Department of Gynecologic Oncology, Jiangxi Cancer Hospital, Nanchang, Jiangxi, China. drgaojun@yeah.net. AD - Nanchang Key Laboratory of Precision Therapy for Gynecological Neoplasms, Nanchang, Jiangxi, China. drgaojun@yeah.net. FAU - Zhu, Hong AU - Zhu H AD - Department of Gynecologic Oncology, Jiangxi Cancer Hospital, Nanchang, Jiangxi, China. 24zhuhong@163.com. LA - eng GR - 82060477/National Natural Science Foundation of China/ GR - 20202BAB206054/Natural Science Foundation of Jiangxi Province/ GR - 202120072/Science and Technology Plan of Jiangxi Provincial Health Commission in 2021/ GR - Hongke Zi (2019) No.228-15/Nanchang Advantage science and technology Innovation team construction Plan project/ GR - Hongke Zi (2021) No. 156-29/Nanchang high-level scientific and technological innovation talents"Double Hundred Plan"project/ GR - 2021B666/General project of Traditional Chinese Medicine Science and Technology Plan of Jiangxi Province in 2021/ GR - Hongke Zi (2019) No.258-6/Nanchang Science and Technology Bureau Science and technology support project/ GR - Hongke Zi (2021) No.129-1/Nanchang City medical health science and technology support project key project/ GR - YC2022-B069/iangxi Province Graduate Students Innovation Special Fund project in 2022(Grant numbers/ PT - Journal Article DEP - 20230616 PL - Netherlands TA - Mol Biol Rep JT - Molecular biology reports JID - 0403234 RN - Q20Q21Q62J (Cisplatin) RN - 0 (4-phenylbutylamine) RN - 49DFR088MY (Platinum) RN - N7TD9J649B (naringin) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - EC 2.7.1.- (Phosphatidylinositol 3-Kinases) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - 0 (Apoptosis Regulatory Proteins) SB - IM MH - Female MH - Humans MH - *Cisplatin/pharmacology MH - Carcinoma, Ovarian Epithelial/drug therapy MH - Platinum MH - Proto-Oncogene Proteins c-akt MH - Phosphatidylinositol 3-Kinases MH - Drug Resistance, Neoplasm MH - Cell Line, Tumor MH - *Ovarian Neoplasms/drug therapy/genetics/metabolism MH - TOR Serine-Threonine Kinases/metabolism MH - Apoptosis MH - Apoptosis Regulatory Proteins MH - Autophagy MH - Cell Proliferation MH - Endoplasmic Reticulum Stress OTO - NOTNLM OT - Autophagy OT - Cisplatin-resistant ovarian cancer OT - Endoplasmic reticulum stress OT - Naringin EDAT- 2023/06/16 13:10 MHDA- 2023/07/31 06:42 CRDT- 2023/06/16 11:10 PHST- 2023/03/30 00:00 [received] PHST- 2023/05/30 00:00 [accepted] PHST- 2023/07/31 06:42 [medline] PHST- 2023/06/16 13:10 [pubmed] PHST- 2023/06/16 11:10 [entrez] AID - 10.1007/s11033-023-08558-3 [pii] AID - 10.1007/s11033-023-08558-3 [doi] PST - ppublish SO - Mol Biol Rep. 2023 Aug;50(8):6457-6468. doi: 10.1007/s11033-023-08558-3. Epub 2023 Jun 16.