School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen, Guangdong 518055, P.R. China.
Science and Education Department, Shenzhen Samii Medical Center, Shenzhen, Guangdong 518118, P.R. China.
Oncol Rep. 2020 Oct;44(4):1596-1604. doi: 10.3892/or.2020.7695. Epub 2020 Jul 17.
The aim of the present study was to explore the antitumor effects of sinoporphyrin sodium (DVDMS)‑mediated photodynamic therapy (PDT) and sonodynamic therapy (SDT) in glioma, and to reveal the underlying mechanisms. The uptake of DVDMS by U‑118 MG cells was detected by flow cytometry (FCM). A 630‑nm semiconductor laser and 1‑MHz ultrasound were used to perform PDT and SDT, respectively. Cell proliferation and apoptosis were evaluated using the Cell Counting Kit‑8 assay, FCM and Hoechst 33258 staining, respectively. Western blot analysis was used to detect protein expression and phosphorylation levels. BALB/c nude mice were used to establish a xenograft model of U‑118 MG cells. DVDMS was injected intravenously and PDT and SDT were performed 24 h later. An in vivo imaging system was used to evaluate the fluorescence of DVDMS, to measure tumor sizes, and to evaluate the therapeutic effects. The uptake of DVDMS by U‑118 MG cells was optimal after 4 h. PDT and SDT following DVDMS injection significantly inhibited the proliferation and increased apoptosis of glioma cells in vitro (P<0.05, P<0.01) respectively. In vivo, the fluorescence intensity of DVDMS was lower in the PDT and SDT groups compared with the DVDMS group, while tumor cell proliferation and weight were lower in the PDT and SDT groups than in the control group (P<0.05, P<0.01). However, there was no significant difference when laser, ultrasound or DVDMS were applied individually, compared with the control group. Hematoxylin and eosin staining suggested that both PDT and SDT induced significant apoptosis and vascular obstruction in cancer tissues. DVDMS‑mediated PDT and SDT inhibited the expression levels of proliferating cell nuclear antigen (PCNA) and Bcl‑xL, increased cleaved ‑caspase 3 levels, and decreased the protein phosphorylation of the PI3K/AKT/mTOR signaling pathway. Changes in the expression of PCNA, and Bcl‑xL and in the levels of cleaved‑caspase 3 were partly reversed by N‑acetyl‑L‑cysteine, a reactive oxygen species (ROS) scavenger. Similar results were obtained with FCM. DVDMS‑mediated PDT and SDT inhibited glioma cell proliferation and induced cell apoptosis in vitro and in vivo, potentially by increasing the generation of ROS and affecting protein expression and phosphorylation levels.
本研究旨在探讨血卟啉单甲醚(DVDMS)介导的光动力疗法(PDT)和声动力疗法(SDT)在胶质瘤中的抗肿瘤作用,并揭示其潜在机制。采用流式细胞术(FCM)检测 U-118 MG 细胞对 DVDMS 的摄取。分别采用 630nm 半导体激光和 1MHz 超声进行 PDT 和 SDT。采用细胞计数试剂盒-8 检测细胞增殖,FCM 和 Hoechst 33258 染色检测细胞凋亡,Western blot 分析检测蛋白表达和磷酸化水平。BALB/c 裸鼠用于建立 U-118 MG 细胞的异种移植模型。静脉注射 DVDMS 24 小时后进行 PDT 和 SDT。采用活体成像系统评估 DVDMS 的荧光,测量肿瘤大小,并评价治疗效果。U-118 MG 细胞摄取 DVDMS 4 小时后达到最佳水平。PDT 和 SDT 后,DVDMS 注射显著抑制了体外胶质瘤细胞的增殖和凋亡(P<0.05,P<0.01)。在体内,与 DVDMS 组相比,PDT 和 SDT 组的 DVDMS 荧光强度较低,与对照组相比,PDT 和 SDT 组的肿瘤细胞增殖和重量较低(P<0.05,P<0.01)。然而,与对照组相比,单独应用激光、超声或 DVDMS 时,差异无统计学意义。苏木精-伊红染色提示 PDT 和 SDT 均诱导了显著的肿瘤组织凋亡和血管阻塞。DVDMS 介导的 PDT 和 SDT 抑制了增殖细胞核抗原(PCNA)和 Bcl-xL 的表达水平,增加了 cleaved-caspase 3 水平,并降低了 PI3K/AKT/mTOR 信号通路的蛋白磷酸化水平。用活性氧(ROS)清除剂 N-乙酰-L-半胱氨酸部分逆转了 PCNA、Bcl-xL 和 cleaved-caspase 3 的表达变化。FCM 也得到了类似的结果。DVDMS 介导的 PDT 和 SDT 抑制了胶质瘤细胞的增殖,并在体外和体内诱导了细胞凋亡,可能是通过增加 ROS 的产生和影响蛋白表达和磷酸化水平来实现的。