背景
顺铂 (DDP) 耐药性在卵巢癌 (OC) 患者中普遍存在，并导致预后不良。因此，开发新的药物干预甚至逆转OC的DDP耐药性具有重要意义。川楝素 (TSN) 是一种从川楝树皮或果实中提取的三萜类化合物，已被证明具有显着的抗肿瘤活性。然而，TSN 对 OC 中 DDP 耐药性的疗效尚未见报道。
目的
本研究的目的是研究TSN对OC对DDP耐药性的影响，并探索体外和体内的分子机制。
方法
使用人 OC 细胞系 (SKOV3) 和 DDP 抗性细胞系 (SKOV3/DDP)。通过CCK-8和集落形成测定法测量细胞增殖。Annexin V/PI双染和hoechst 33342核染色检测细胞凋亡。Transwell和伤口愈合试验分别用于确定细胞的侵袭和迁移潜力。采用实时定量PCR（qPCR）和western blotting检测miR-195/ERK/β-catenin通路相关分子的表达。使用异种移植模型、TUNEL 染色试验和免疫组织化学研究了 TSN 在体内对 OC 的 DDP 抗性的影响和机制。
结果
TSN 在体外和体内提高了 SKOV3/DDP 细胞的 DDP 敏感性，体现在促进抑制增殖、侵袭、迁移和上皮间充质转化 (EMT) 以及诱导细胞凋亡。TSN 可以通过上调 miR-195-5p 表达然后抑制在 SKOV3/DDP 细胞中激活的 ERK/GSK3β/β-catenin 通路来调节 miR-195/ERK/β-catenin 轴。此外，β-连环蛋白通路激活剂 LiCl 或 miR-195-5p 沉默的共同处理部分恢复了先前被 TSN 抑制的 DDP 抗性。
结论
体外和体内数据均表明，TSN 可通过调节 miR-195/ERK/β-catenin 通路降低 OC 中的 DDP 耐药性，突出了 TSN 作为有利于克服 OC 中临床 DDP 耐药性的有效药物的潜力。
"点击查看英文标题和摘要"
Toosendanin reduces cisplatin resistance in ovarian cancer through modulating the miR-195/ERK/β-catenin pathway
Background
Cisplatin (DDP) resistance is prevalent in ovarian cancer (OC) patients and contributes to the poor prognosis. Therefore, it is of great significance to develop new agent to intervene and even reverse DDP resistance in OC. Toosendanin (TSN), a triterpenoid extracted from the bark or fruits of Melia toosendan Sieb et Zucc, has been proved to possess significant antitumor activities. However, the efficacy of TSN on DDP resistance in OC has not been reported yet.
Purpose
The aim of this study is to investigate the effects of TSN on DDP resistance in OC and explore the molecular mechanism in vitro and in vivo.
Methods
Human OC cell line (SKOV3) and DDP-resistant cell line (SKOV3/DDP) were used. Cell proliferation was measured by CCK-8 and colony formation assay. Annexin V/PI double staining and hoechst 33342 nuclear staining were employed to detect cell apoptosis. Transwell and wound-healing assay were used to determine the invasion and migration potential of cells respectively. Quantitative real-time PCR (qPCR) and western blotting were performed to detect the expression of molecules related to miR-195/ERK/β-catenin pathway. The effects and mechanism of TSN on DDP resistance of OC in vivo was investigated using xenograft model, TUNEL staining assay and immunohistochemistry.
Results
TSN improved the DDP sensitivity of SKOV3/DDP cells in vitro and in vivo, reflected in promoting inhibition of proliferation, invasion, migration and epithelial mesenchymal transformation (EMT) as well as induction of apoptosis. TSN could modulate the miR-195/ERK/β-catenin axis by upregulating the miR-195-5p expression and then suppressing ERK/GSK3β/β-catenin pathway which were activated in SKOV3/DDP cells. Moreover, co-treatment of β-catenin pathway activator LiCl or miR-195-5p silencing partially recovered the DDP resistance which was previously repressed by TSN.
Conclusion
Both in vitro and in vivo data demonstrated that TSN could reduce DDP resistance in OC through regulating the miR-195/ERK/β-catenin pathway, highlighting the potential of TSN as an effective agent for favoring overcoming clinical DDP resistance in OC.
京公网安备 11010802027423号