Zihao Zhao , Qihong Wu , Yangyang Xu , Yuhuan Qin , Runsang Pan , Qingqi Meng , Siming Li
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Cell scratch assay, CCK-8, FCM, and the EdU assay were utilized to evaluate the effect of groenlandicine on cisplatin-resistant cells. WB, immunofluorescence, and PCR were conducted to measure the levels of TOP1, Bcl-2, BAX, Caspase-9, and Caspase-3. Additionally, a subcutaneous tumor model was established in nude mice to verify the efficacy of groenlandicine.</p><p>Groenlandicine reduced the migration and proliferation while promoting apoptosis in OS cells, effectively damaging them. Meanwhile, groenlandicine exhibited weak cytotoxicity in 293T cells. Combination with cisplatin enhanced tumor-killing activity, markedly activating BAX, cleaved-Caspase-3, and cleaved-Caspase-9, while inhibiting the Bcl2 pathway in cisplatin-resistant OS cells. Moreover, the level of TOP1, elevated in cisplatin-resistant OS cells, was down-regulated by groenlandicine both <em>in vitro</em> and <em>in vivo</em>. 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引用次数: 0
摘要
格根地新是从黄连中分离出来的一种原小檗碱，黄连是一种广泛使用的传统中药，以其多种生物活性而闻名。本研究采用配体虚拟筛选法（LBVS）和分子对接法来筛选药物。采用配体虚拟筛选（LBVS）法和分子对接法筛选药物，并用CCK-8和FCM测定格仑地新对转染过表达或低表达TOP1慢病毒的OS细胞的影响。细胞划痕试验、CCK-8、FCM 和 EdU 试验用于评估格列宁对顺铂耐药细胞的影响。WB、免疫荧光和 PCR 检测了 TOP1、Bcl-2、BAX、Caspase-9 和 Caspase-3 的水平。此外，还建立了裸鼠皮下肿瘤模型来验证格仑地新的疗效。格仑地新在促进 OS 细胞凋亡的同时减少了其迁移和增殖，有效地破坏了 OS 细胞。同时，格陵兰碱对 293T 细胞的细胞毒性较弱。与顺铂联用可增强肿瘤杀伤活性，显著激活顺铂耐药 OS 细胞中的 BAX、裂解-Caspase-3 和裂解-Caspase-9，同时抑制 Bcl2 通路。此外，在顺铂耐药的OS细胞中升高的TOP1水平在体外和体内均被格罗恩地辛下调。动物实验证实，格罗兰地辛与顺铂联用可抑制OS的生长，且肾毒性较低。格罗兰地辛通过BAX/Bcl-2/Caspase-9/Caspase-3途径诱导细胞凋亡，并增强耐药OS细胞对顺铂的敏感性。因此，格罗兰地辛有望成为逆转顺铂治疗耐药性的潜在药物。
Groenlandicine enhances cisplatin sensitivity in cisplatin-resistant osteosarcoma cells through the BAX/Bcl-2/Caspase-9/Caspase-3 pathway
Groenlandicine is a protoberberine alkaloid isolated from Coptidis Rhizoma, a widely used traditional Chinese medicine known for its various biological activities. This study aims to validate groenlandicine’s effect on both cisplatin-sensitive and cisplatin-resistant osteosarcoma (OS) cells, along with exploring its potential molecular mechanism.
The ligand-based virtual screening (LBVS) method and molecular docking were employed to screen drugs. CCK-8 and FCM were used to measure the effect of groenlandicine on the OS cells transfected by lentivirus with over-expression or low-expression of TOP1. Cell scratch assay, CCK-8, FCM, and the EdU assay were utilized to evaluate the effect of groenlandicine on cisplatin-resistant cells. WB, immunofluorescence, and PCR were conducted to measure the levels of TOP1, Bcl-2, BAX, Caspase-9, and Caspase-3. Additionally, a subcutaneous tumor model was established in nude mice to verify the efficacy of groenlandicine.
Groenlandicine reduced the migration and proliferation while promoting apoptosis in OS cells, effectively damaging them. Meanwhile, groenlandicine exhibited weak cytotoxicity in 293T cells. Combination with cisplatin enhanced tumor-killing activity, markedly activating BAX, cleaved-Caspase-3, and cleaved-Caspase-9, while inhibiting the Bcl2 pathway in cisplatin-resistant OS cells. Moreover, the level of TOP1, elevated in cisplatin-resistant OS cells, was down-regulated by groenlandicine both in vitro and in vivo. Animal experiments confirmed that groenlandicine combined with cisplatin suppressed OS growth with lower nephrotoxicity.
Groenlandicine induces apoptosis and enhances the sensitivity of drug-resistant OS cells to cisplatin via the BAX/Bcl-2/Caspase-9/Caspase-3 pathway. Groenlandicine inhibits OS cells growth by down-regulating TOP1 level.Therefore, groenlandicine holds promise as a potential agent for reversing cisplatin resistance in OS treatment.
期刊介绍：
The Journal of Bone Oncology is a peer-reviewed international journal aimed at presenting basic, translational and clinical high-quality research related to bone and cancer.
As the first journal dedicated to cancer induced bone diseases, JBO welcomes original research articles, review articles, editorials and opinion pieces. Case reports will only be considered in exceptional circumstances and only when accompanied by a comprehensive review of the subject.
The areas covered by the journal include:
Bone metastases (pathophysiology, epidemiology, diagnostics, clinical features, prevention, treatment)
Preclinical models of metastasis
Bone microenvironment in cancer (stem cell, bone cell and cancer interactions)
Bone targeted therapy (pharmacology, therapeutic targets, drug development, clinical trials, side-effects, outcome research, health economics)
Cancer treatment induced bone loss (epidemiology, pathophysiology, prevention and management)
Bone imaging (clinical and animal, skeletal interventional radiology)
Bone biomarkers (clinical and translational applications)
Radiotherapy and radio-isotopes
Skeletal complications
Bone pain (mechanisms and management)
Orthopaedic cancer surgery
Primary bone tumours
Clinical guidelines
Multidisciplinary care
Keywords: bisphosphonate, bone, breast cancer, cancer, CTIBL, denosumab, metastasis, myeloma, osteoblast, osteoclast, osteooncology, osteo-oncology, prostate cancer, skeleton, tumour.
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