Division of Nephrology, Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan.
Department of Obstetrics and Gynecology, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan.
Int J Mol Sci. 2023 Feb 8;24(4):3353. doi: 10.3390/ijms24043353.
This study investigated whether melatonin (Mel) would promote cisplatin to suppress the proliferation and growth of bladder cancer (BC) cells by inhibiting cellular prion protein (PrP)-mediated cell stress and cell proliferation signaling. An immunohistochemical staining of tissue arrays from BC patients demonstrated that the PrP expression was significantly upregulated from stage I to III BC ( < 0.0001). The BC cellline of T24 was categorized into G1 (T24), G2 (T24 + Mel/100 μM), G3 (T24+cisplatin/6 μM), G4 (PrP overexpression in T24 (i.e., PrP-T24)), G5 (PrP-T24+Mel), and G6 (PrP-T24+cisplatin). When compared with a human uroepithelial cell line (SV-HUC-1), the cellular viability/wound healing ability/migration rate were significantly increased in T24 cells (G1) and further significantly increased in PrP-T24 cells (G4); and they were suppressed in Mel (G2/G5) or cisplatin (G3/G6) treatment (all < 0.0001). Additionally, the protein expressions of cell proliferation (PI3K/p-Akt/p-m-TOR/MMP-9/PrP), cell cycle/mitochondrial functional integrity (cyclin-D1/clyclin-E1/ckd2/ckd4/mitochondrial-cytochrome-C/PINK1), and cell stress (RAS/c-RAF/p-MEK1/2, p-ERK1/2) markers showed a similar pattern of cell viability among the groups (all < 0.001). After the BC cell line of UMUC3 was implanted into nude mouse backs, by day 28 mthe BC weight/volume and the cellular levels of PrP/MMP-2/MMP-9 were significantly, gradually reduced from groups one to four (all < 0.0001). The protein expressions of cell proliferation (PI3K/p-Akt/p-m-TOR/MMP-9/PrP), cell cycle/mitophagy (cyclin-D1/clyclin-E1/ckd2/ckd4/PINK1), and cell stress (RAS/c-RAF/p-MEK1,2/p-ERK1,2) signaling were significantly, progressively reduced from groups one to four, whereas the protein expressions of apoptotic (Mit-Bax/cleaved-caspase-3/cleaved-PARP) and oxidative stress/mitochondrial damaged (NOX-1/NOX-2/cytosolic-cytochrome-C/p-DRP1) markers expressed an opposite pattern of cell proliferation signaling among the groups (all < 0.0001). Mel-cisplatin suppressed BC cell growth/proliferation via inhibiting the PrP in upregulating the cell proliferation/cell stress/cell cycle signaling.
本研究旨在探讨褪黑素（Mel）是否通过抑制细胞朊蛋白（PrP）介导的细胞应激和细胞增殖信号来促进顺铂抑制膀胱癌（BC）细胞的增殖和生长。对 BC 患者组织阵列的免疫组织化学染色表明，PrP 的表达从 I 期到 III 期 BC 显著上调（<0.0001）。T24 的 BC 细胞系被分为 G1（T24）、G2（T24+Mel/100 μM）、G3（T24+顺铂/6 μM）、G4（PrP 在 T24 中的过表达（即 PrP-T24））、G5（PrP-T24+Mel）和 G6（PrP-T24+顺铂）。与人类尿路上皮细胞系（SV-HUC-1）相比，T24 细胞（G1）的细胞活力/伤口愈合能力/迁移率显著增加，PrP-T24 细胞（G4）的细胞活力/伤口愈合能力/迁移率进一步显著增加；而 Mel（G2/G5）或顺铂（G3/G6）处理则抑制了它们的增殖（均<0.0001）。此外，细胞增殖（PI3K/p-Akt/p-m-TOR/MMP-9/PrP）、细胞周期/线粒体功能完整性（cyclin-D1/clyclin-E1/ckd2/ckd4/线粒体细胞色素-C/PINK1）和细胞应激（RAS/c-RAF/p-MEK1/2、p-ERK1/2）标志物的蛋白表达在各组之间表现出相似的细胞活力模式（均<0.001）。当将 UMUC3 的 BC 细胞系植入裸鼠背部后，到第 28 天，BC 的重量/体积和细胞内 PrP/MMP-2/MMP-9 的水平从第 1 组到第 4 组逐渐降低（均<0.0001）。细胞增殖（PI3K/p-Akt/p-m-TOR/MMP-9/PrP）、细胞周期/自噬（cyclin-D1/clyclin-E1/ckd2/ckd4/PINK1）和细胞应激（RAS/c-RAF/p-MEK1,2/p-ERK1,2）信号的蛋白表达从第 1 组到第 4 组逐渐降低，而凋亡（Mit-Bax/cleaved-caspase-3/cleaved-PARP）和氧化应激/线粒体损伤（NOX-1/NOX-2/cytosolic-cytochrome-C/p-DRP1）标志物的蛋白表达则表现出与细胞增殖信号相反的模式（均<0.0001）。Mel-顺铂通过抑制 PrP 上调细胞增殖/细胞应激/细胞周期信号来抑制 BC 细胞生长/增殖。
