The Effect of Triptolide-Loaded Exosomes on the Proliferation and Apoptosis of Human Ovarian Cancer SKOV3 Cells
The Effect of Triptolide-Loaded Exosomes on the Proliferation and Apoptosis of Human Ovarian Cancer SKOV3 Cells
复制标题
雷公藤甲素外泌体对人卵巢癌SKOV3细胞增殖和凋亡的影响
DOI：
10.1155/2019/2595801
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发表时间：
2019-01-01
影响因子：
--
通讯作者：
Li, He
中科院分区：
文献类型：
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作者：
Liu, Huan;Shen, Ming;Li, He
雷公藤内酯醇具有抗肿瘤作用，但其水溶性差、肝毒性和肾毒性限制了其临床应用。本研究构建了雷公藤甲素外泌体给药系统（TP-Exos），并观察其对SKOV 3细胞增殖和凋亡的影响。通过超离心和超滤离心收集SKOV 3-外泌体（SK-Exos）。通过超声处理和超滤离心构建TP-Exos。SK-Exos和TP-Exos通过透射电子显微镜、蛋白质印迹法、纳米颗粒跟踪分析和高效液相色谱法进行表征。采用细胞摄取exosomes、MTT法、BrdU法和细胞凋亡实验研究TP-Exos对卵巢癌细胞的作用。采用肿瘤靶向性研究、小鼠肿瘤体积监测和TdT介导的dUTP缺口末端标记（TUNEL）法评价TP-Exos对卵巢癌的体内作用。通过肝和肾功能以及主要器官（心、肝、脾、肺、肾和卵巢）的组织病理学评价TP-Exos的体内毒性。结果表明，TP-Exos不仅具有exosomes的一般特征，而且具有较高的药物包封率。PKH 26标记的exosomes（PKH 26-Exos）可被SKOV 3细胞摄取，Dir标记的exosomes（Dir-Exos）可富集到荷瘤小鼠的肿瘤部位。TP-Exos对SKOV 3细胞的细胞毒作用和凋亡作用均弱于游离TP，而对肿瘤细胞增殖和肿瘤生长的抑制作用强于游离TP。此外，TP-Exos对肝脏和脾脏有毒性作用。总之，TP-Exos可能是卵巢癌的一种有前途的策略，但它们需要进一步优化以减轻对肝脏和脾脏的损害。
Triptolide has been proven to possess anticancer efficacy; however, its application in the clinical practice was limited by poor water solubility, hepatotoxicity, and nephrotoxicity. In this study, a triptolide-loaded exosomes delivery system (TP-Exos) was constructed and its effects on the proliferation and apoptosis of SKOV3 cells in vitro and in vivo were observed. SKOV3-exosomes (SK-Exos) were collected by ultracentrifugation and ultrafiltration centrifugation. TP-Exos was constructed by sonication and ultrafiltration centrifugation. SK-Exos and TP-Exos were characterized by transmission electron microscopy, western blotting, nanoparticle-tracking analysis, and high-performance liquid chromatography. Cellular uptake of exosomes, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, bromodeoxyuridine (BrdU) cell proliferation assay, and cell apoptosis experiment were used to study the effect of TP-Exos on ovarian cancer in vitro. Tumor-targeting study of exosomes, monitoring the tumor volume of mice, and TdT-mediated dUTP Nick-End labeling (TUNEL) assay were used to evaluate the effect of TP-Exos on ovarian cancer in vivo. The toxicity of TP-Exos in vivo was evaluated by liver and kidney function and histopathology of major organs (heart, liver, spleen, lung, kidney, and ovary). The results revealed that TP-Exos not only have the general characteristics of exosomes but also have high drug encapsulation efficiency. Besides, PKH26 labeled exosomes (PKH26-Exos) could be uptaken by SKOV3 cells, and Dir labeled exosomes (Dir-Exos) could be enriched to the tumor site of tumor bearing mice. Furthermore, the cytotoxic and apoptotic effects on SKOV3 cells of TP-Exos were weaker than those of free TP, and tumor cell proliferation inhibition and tumor growth inhibition were stronger than that of free TP. Moreover, TP-Exos have toxic effect on liver and spleen. In conclusion, the TP-Exos could be a promising strategy for ovarian cancer, but they need to be further optimized to attenuate the damage to liver and spleen.