生物技术进展 ›› 2021, Vol. 11 ›› Issue (2): 244-251.DOI: 10.19586/j.2095-2341.2020.0121

• 研究论文 • 上一篇    

负载雷公藤甲素的TPGS-b-(PCL-ran-PGA)纳米制剂体外抑制宫颈癌细胞生长的研究

郑义1,郑静薇2,周煜玲1,梁欣1,包君丽1,杨菊红1,曾凡杞1*   

  1. 1.中国科学院大学深圳医院医学实验中心, 广东 深圳 518106;
    2.吉林大学临床医学院, 长春 220100
  • 收稿日期:2020-09-30 出版日期:2021-03-25 发布日期:2020-11-04
  • 通讯作者: 曾凡杞 E-mail: 332941241@qq.com
  • 作者简介:郑义 E-mail: zhengyi1205@126.com
  • 基金资助:
    广东省中医药局科研项目（20191287）；深圳市光明区科技局项目（GM2019010003）。

In vitro Study of the Inhibition Growth of Cervical Cancer Cells by Triptolide-loading TPGS-b-(PCL-ran-PGA) Nanoparticles

ZHENG Yi, ZHENG Jingwei, ZHOU Yuling, LIANG Xin, BAO Junli, YANG Juhong, ZENG Fanqi   

  1. 1.Center for Medical Experiments（CME）, University of Chinese Academy of Sciences-Shenzhen Hospital, Guangdong Shenzhen 518106, China；
    2.Clinical Medical School of Jilin University, Changchun 220100,  China
  • Received:2020-09-30 Online:2021-03-25 Published:2020-11-04

PDF

摘要： 雷公藤甲素（triptolide，TPL）是传统中药雷公藤的主要活性成分，具有抗炎、抗肿瘤活性，但其毒副作用限制了临床上的广泛使用。为了探讨以TPGS-b-(PCL-ran-PGA)为载体制备的TPGS-b-(PCL-ran-PGA)/TPL纳米粒的表征和体外对宫颈癌细胞的抑制作用，采用乳化/溶剂挥发法，优化TPGS-b-(PCL-ran-PGA)与TPL比例，制备TPGS-b-(PCL-ran-PGA)/TPL纳米粒，对纳米粒进行表征，包括粒径大小、ζ电位、包封率、累积释放率，用MTS法体外研究游离型TPL和TPGS-b-(PCL-ran-PGA)/TPL纳米粒对宫颈癌细胞半数抑制浓度（IC50），用克隆形成实验分析TPGS-b-(PCL-ran-PGA)/TPL纳米粒对宫颈癌细胞HeLa的抑制作用，用流式细胞仪分析纳米粒对HeLa细胞凋亡的影响。结果显示：当TPGS-b-(PCL-ran-PGA)与TPL为50∶1时制备的纳米粒粒径为(95.3±5.2)nm,zeta电位为(-12.2±0.9)mV,其累积释放曲线呈双相分布，TPGS-b-(PCL-ran-PGA)纳米粒对HeLa细胞在24、48和72 h的IC50（2.8、1.8、0.9 μg·L-1）远远低于游离型TPL（P<0.01）,克隆形成实验证明纳米粒能显著抑制肿瘤细胞生长，并能显著诱导HeLa细胞凋亡。研究结果表明，TPGS-b-(PCL-ran-PGA)/TPL纳米粒能抑制宫颈癌细胞HeLa的生长，其作用主要通过TPL和TPGS共同诱导细胞凋亡，可以作为抗宫颈癌等肿瘤的候选药物。

关键词: 雷公藤甲素, 纳米粒, 宫颈癌, TPGS-b-(PCL-ran-PGA)

Abstract: Triptolide(TPL), as a main compound isolated from traditional Chinese medicine Tripterygium wilfordii, has a broad range of bioactivities including anti inflammation and anticancer activity. However, its clinical application of TPL is restricted by its adverse effects. To explore the characteristics of TPGS-b-(PCL-ran-PGA)/TPL nanoparticles（NPs） and functions as anti-cervical cancer cells in vitro, TPGS-b-(PCL-ran-PGA)/TPL NPs were prepared by using emulsion/solvent evaporation method and optimizing the ratio of TPL and TPGS-b-(PCL-ran-PGA), and their size distribution, morphology, zeta potential, drug encapsulation efficiency, drug-release profile, were characterized. The 50% inhibitory concentration(IC50) and inhibition effect of both free TPL and TPGS-b-(PCL-ran-PGA)/TPL NPs on cervical cancer cells were measured by MTS method and colony formation assay, respectively. By using flow cytometer, HeLa cell apoptosis induced by TPGS-b-(PCL-ran-PGA)/TPL NPs was detected. Results showed that, TPGS-b-(PCL-ran-PGA)/TPL NPs were prepared at the 50∶1 ratio of TPGS-b-(PCL-ran-PGA)and TPL. The size of TPGS-b-(PCL-ran-PGA)/TPL NPs was (95.3±5.2) nm with a mean zeta potential (-12.2±0.9) mV.  A bi-phase drug-release profile from these NPs was observed at pH 7.4 in drug release studies. TPGS-b-(PCL-ran-PGA)/TPL NPs（IC50 2.8,1.8, 0.9 μg·L-1 at 24, 48 and 72 h, respectively）were proved more potent against HeLa cells than free TPL (IC50  232,12.5 and 6.8 at 24, 48 and 72 h, respectively). TPGS-b-(PCL-ran-PGA)/TPL NPs could significantly inhibit tumor cell colony formation and induce apoptosis compared with free TPL. The results indicated that, TPGS-b-(PCL-ran-PGA)/TPL NPs could inhibit the cancer cell growth in vitro, since in combination of TPL and TPGS leads to apoptosis. It suggested that TPGS-b-(PCL-ran-PGA)/TPLNPs can serve as candidate agents against cancers including cervical cancer.

Key words: triptolide(TPL), nanoparticles(NPs), cervical cancer, TPGS-b-(PCL-ran-PGA)

版权所有 © 2021《生物技术进展》编辑部

京ICP备07026971-2 　　 京公网安备11010802021232号

本系统由北京玛格泰克科技发展有限公司设计开发 技术支持：support@magtech.com.cn