Postgraduate Training Base Alliance of Wenzhou Medical University (Zhejiang Cancer Hospital), Hangzhou, 310022, Zhejiang, China.
Traditional Chinese Medicine Pharmacy, Zhejiang Hospital, Hangzhou, 310022, Zhejiang, China.
Sci Rep. 2024 Sep 10;14(1):21130. doi: 10.1038/s41598-024-72198-1.
Morphine has been suggested to affect cancer cell dynamics and decrease survival rates in lung cancer patients at specific doses, but the precise mechanisms poorly understood. In this study, we aimed to investigate the molecular mechanisms by which morphine modulates the malignant characteristics of non-small cell lung cancer. Cell proliferation was assessed via the Cell Counting Kit-8 assay, and cell migration and invasion were examined via wound healing and Transwell assays. We employed immunofluorescence staining to evaluate E-cadherin expression in A549 and Lewis lung cancer (LLC) cell lines and immunohistochemistry to evaluate E-cadherin expression in nude mice tumours. Additionally, the in vivo effects of morphine on lung cancer progression were explored in a xenograft tumour experiments, in which naloxone was used as a morphine antagonist. Western blot analysis was performed to detect E-cadherin, phosphorylated mTOR (p-mTOR), mTOR, phosphorylated AKT (p-AKT), AKT, phosphorylated PI3K (p-PI3K), and PI3K protein levels in A549 and LLC cells as well as in tumour samples. Morphine (10 µM) significantly increased the proliferation of A549 and LLC cells in vitro (p < 0.05). It also enhanced the migratory and invasive capacities of these cell lines (p < 0.01). Mechanistically, morphine treatment (10 µM) led to a reduction in the expression of E-cadherin, and an increase in the phosphorylation of PI3K, AKT, and mTOR in A549 and LLC cells (p < 0.01). Morphine treatment (1.5 mg/kg) also reduced E-cadherin expression in xenograft tumours and promoted tumour growth in vivo (p < 0.05). This effect was reversed by naloxone (0.1 mg/kg). The results demonstrated that morphine stimulates the malignant proliferation of A549 and LLC cell lines and promotes xenograft tumour growth. Perhaps by specifically targeting MOR, morphine triggers a signalling cascade that activates the PI3K/AKT/mTOR pathway while inhibiting the EMT marker E-cadherin, which may consequently promote the progression of lung cancer.
吗啡已被证实会影响特定剂量下肺癌患者的癌细胞动力学并降低其存活率,但确切的机制尚不清楚。在这项研究中,我们旨在研究吗啡调节非小细胞肺癌恶性特征的分子机制。通过细胞计数试剂盒-8 检测细胞增殖,通过划痕愈合和 Transwell 检测评估细胞迁移和侵袭。我们采用免疫荧光染色评估 A549 和 Lewis 肺癌(LLC)细胞系中 E-钙黏蛋白的表达,采用免疫组化评估裸鼠肿瘤中 E-钙黏蛋白的表达。此外,在异种移植肿瘤实验中探索了吗啡对肺癌进展的体内影响,其中纳洛酮被用作吗啡拮抗剂。Western blot 分析用于检测 A549 和 LLC 细胞以及肿瘤样本中 E-钙黏蛋白、磷酸化 mTOR(p-mTOR)、mTOR、磷酸化 AKT(p-AKT)、AKT、磷酸化 PI3K(p-PI3K)和 PI3K 蛋白水平。吗啡(10 μM)显著增加了 A549 和 LLC 细胞的体外增殖(p<0.05)。它还增强了这些细胞系的迁移和侵袭能力(p<0.01)。在机制上,吗啡处理(10 μM)导致 A549 和 LLC 细胞中 E-钙黏蛋白表达减少,PI3K、AKT 和 mTOR 磷酸化增加(p<0.01)。吗啡处理(1.5mg/kg)也降低了异种移植肿瘤中的 E-钙黏蛋白表达并促进了体内肿瘤生长(p<0.05)。纳洛酮(0.1mg/kg)逆转了这种作用。结果表明,吗啡刺激 A549 和 LLC 细胞系的恶性增殖,并促进异种移植肿瘤的生长。也许通过特异性靶向 MOR,吗啡触发了一个信号级联,激活了 PI3K/AKT/mTOR 通路,同时抑制 EMT 标志物 E-钙黏蛋白,这可能促进肺癌的进展。