Sinoporphyrin Sodium-Mediated Sonodynamic Therapy Inhibits RIP3 Expression and Induces Apoptosis in the H446 Small Cell Lung Cancer Cell Line.
Sinoporphyrin Sodium-Mediated Sonodynamic Therapy Inhibits RIP3 Expression and Induces Apoptosis in the H446 Small Cell Lung Cancer Cell Line.
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华卟啉钠介导的声动力疗法抑制 H446 小细胞肺癌细胞系中 RIP3 的表达并诱导细胞凋亡。
DOI：
10.1159/000496045
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发表时间：
2018
影响因子：
--
通讯作者：
Yu Yan
中科院分区：
文献类型：
--
作者：
Shen Jing;Cao Shoubo;Sun Xin;Pan Bo;Cao Jingyan;Che Dehai;Jin Shi;Cao Yingyue;Tian Ye;Yu Yan
背景/目的超声动力治疗（SDT）有望成为解决肺癌晚期转移患者临床难题的新方法。超声波具有非侵入性和深穿透性的优点。本研究探讨了SDT联合新型声敏剂sinoporphyrin sodium （DVDMS）对人小细胞肺癌H446细胞株的体外和体内抗肿瘤作用。方法用荧光分光光度计检测DVDMS的吸收，用细胞计数试剂盒-8测定DVDMS的毒性。采用JC-1荧光探针测定线粒体膜电位（MMP）。流式细胞术检测细胞凋亡，western blotting检测细胞凋亡相关蛋白。细胞因子的表达采用酶联免疫吸附法和实时定量PCR检测。为了验证体外结果，我们检测了DVDMS-SDT后异种移植裸鼠模型的肿瘤体积和重量变化。采用苏木精和伊红染色观察小鼠肿瘤、心、肝、脾、肺、肾的变化，免疫组化检测肿瘤CD34和受体相互作用蛋白激酶-3 （RIP3）的表达变化，末端脱氧核苷酸转移酶介导的dUTP镍端标记观察肿瘤组织的凋亡。结果dvdms - sdt处理的H446细胞细胞凋亡率和活性氧（ROS）、裂解型caspase-3、裂解型caspase-8、裂解型caspase-9、caspase-10水平升高，MMP、RIP3、b细胞淋巴瘤2、血管内皮生长因子、肿瘤坏死因子-α水平降低。声毒性作用由活性氧介导，并被活性氧清除剂（n -乙酰- l-半胱氨酸）降低。在小鼠体内异种移植模型中，DVDMS-SDT显示出有效的抗癌作用，无明显副作用。结论dvdms - sdt诱导H446细胞凋亡，部分通过线粒体介导的凋亡信号通路靶向线粒体，外源性凋亡通路也参与其中。细胞凋亡和RIP3表达的变化都与ROS的产生密切相关。由于DVDMS-SDT的无创特性，它将有利于小细胞肺癌的治疗。
Background/AimsSonodynamic therapy (SDT) is expected to be a new method to solve the clinical problems caused by advanced metastasis in patients with lung cancer. The use of ultrasound has the advantage of being noninvasive, with deep-penetration properties. This study explored the anti-tumor effect of SDT with a new sonosensitizer, sinoporphyrin sodium (DVDMS), on the human small cell lung cancer H446 cell line in vitro and in vivo.MethodsAbsorption of DVDMS was detected by a fluorescence spectrophotometer, and DVDMS toxicity was determined using a Cell Counting Kit-8. Mitochondrial membrane potential (MMP) was assessed using the JC-1 fluorescent probe. Cell apoptosis was measured by flow cytometry, and apoptosis-related proteins were detected by western blotting. The expression of cytokines was measured using an enzyme-linked immunosorbent assay and quantitative real-time PCR. To verify the in vitro results, we detected tumor volumes and weight changes in a xenograft nude mouse model after DVDMS-SDT. Hematoxylin and eosin staining was used to observe changes to the tumor, heart, liver, spleen, lung, and kidney of the mice, and immunohistochemistry was used to examine changes in the expression of tumor CD34 and receptor-interacting protein kinase-3 (RIP3), while terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was used to observe apoptosis in tumor tissues.ResultsDVDMS-SDT-treated H446 cells increased the rate of cellular apoptosis and the levels of reactive oxygen species (ROS), cleaved caspase-3, cleaved caspase-8, cleaved caspase-9, and caspase-10, and decreased the levels of MMP, RIP3, B-cell lymphoma 2, vascular endothelial growth factor, and tumor necrosis factor-α. The sonotoxic effect was mediated by ROS and was reduced by a ROS scavenger (N-acetyl-L-cysteine). In the in vivo mouse xenograft model, DVDMS-SDT showed efficient anti-cancer effects with no visible side effects.ConclusionDVDMS-SDT induced apoptosis in H446 cells, in part by targeting mitochondria through the mitochondria-mediated apoptosis signaling pathway, and the extrinsic apoptosis pathway was also shown to be involved. Both apoptosis and changes in RIP3 expression were closely related to the generation of ROS. DVDMS-SDT will be advantageous for the management of small cell lung cancer due to its noninvasive characteristics.