School of Pharmaceutical Science, Shandong University, Jinan 250012, China.
Institute of Basic Medical Sciences, Qilu Hospital, Shandong University, Jinan 250012, China; The Key Laboratory of Cardiovasular Remodelling and Function Research, Chinese Ministry of Health, Qilu Hospital, Shandong University, Jinan 250012, China.
Acta Biomater. 2017 Oct 15;62:257-272. doi: 10.1016/j.actbio.2017.08.034. Epub 2017 Aug 30.
In this study, novel prodrug-modified cationic liposome nanocomplexes (Combo NCs) were reported for gemcitabine (GEM) and docetaxel (DTX) co-delivery. This nanoplatform exhibited multiple favorable characteristics, such as a 'green' fabrication with a one-step chemical reaction, appropriate size (∼200nm) and distribution (PDI<0.2), low zeta potential (-31.1mv), high drug-loading efficiency (9.3% GEM plus 3.1% DTX, wt%) and pH and enzymatic dual-stimulus-responsive release properties. Immunofluorescence and cellular uptake studies showed that Combo NCs efficiently targeted overexpressed CD44 in MDA-MB-231 carcinoma. In vitro studies revealed that Combo NCs played a critical role in the synergistic induction of cytotoxicity, apoptosis and inhibition of wound healing. Combo NCs were confirmed to exhibit great potency for increasing S phase arrest and remodeling the CDA and dCK balance by decreasing the mRNA expression of CDA down to 0.09-fold and increasing the mRNA expression of dCK by 1.36-fold, remarkably increasing the dCK/CDA ratio to 15.3-fold compared with the blank control. The biodistribution results obtained in vivo revealed an effective accumulation in tumor foci. All of these advantages of Combo NCs contributed to their remarkable anti-tumor efficacy without systemic toxicity as well as their apoptosis-enhancing and anti-proliferative capacities, as determined by TUNEL and Ki67 immunohistochemistry in vivo. Consequently, such a rationally contemplated co-delivery system demonstrated the promising potential of clinical applications for triple-negative breast cancer therapy.
The Combo NCs were innovatively applied for co-delivery of hydrophilic GEM and hydrophobic DTX. The ester bond linking and shielding effect of HA-GEM made the carriers achieve synchronous release properties, which was determined in in vitro release study. Due to the HA modification, the vectors own great potency for positive targeting to CD44 overexpressed triple-negative breast cancer cells MDA-MB-231. Cytotoxicity and apoptosis studies confirmed the targeting effect and synergism between two drugs. Interestingly, we found in cell cycle study, drug combinations (free combination or Combo NCs) didn't show a rise in G2M phase, which was significantly higher when treated DTX alone. We further discovered the role of DTX in combinations may involve in modulating GEM associated enzymes thus enhancing the efficacy of GEM. Consequently, this nanoplatform provided a novel solution for achieving targeted co-delivery and potentiating effect in cancer therapy.
在这项研究中，报告了新型前药修饰的阳离子脂质体纳米复合物（Combo NCs）用于吉西他滨（GEM）和多西他赛（DTX）的共递送。该纳米平台具有多种有利的特性，例如“绿色”制造，一步化学反应，合适的尺寸（约 200nm）和分布（PDI<0.2），低zeta 电位（-31.1mv），高载药效率（9.3% GEM 加 3.1% DTX，wt%）和 pH 和酶双重刺激响应释放特性。免疫荧光和细胞摄取研究表明，Combo NCs 能够有效地靶向 MDA-MB-231 癌中过表达的 CD44。体外研究表明，Combo NCs 在协同诱导细胞毒性、凋亡和抑制伤口愈合方面发挥了关键作用。Combo NCs 被证实通过将 CDA 的 mRNA 表达降低到 0.09 倍并将 dCK 的 mRNA 表达增加 1.36 倍，从而显著增加 dCK/CDA 比值至 15.3 倍，来发挥增加 S 期阻滞和重塑 CDA 和 dCK 平衡的重要作用与空白对照相比。体内分布结果表明，纳米复合物在肿瘤病灶中有有效积聚。所有这些 Combo NCs 的优势都有助于它们在没有全身毒性的情况下表现出显著的抗肿瘤功效，以及通过体内 TUNEL 和 Ki67 免疫组化确定的增强凋亡和抗增殖能力。因此，这种经过深思熟虑的共递药系统为三阴性乳腺癌治疗的临床应用展示了有前途的潜力。
Combo NCs 被创新性地应用于亲水性 GEM 和疏水性 DTX 的共递送。HA-GEM 中的酯键连接和屏蔽作用使载体在体外释放研究中实现了同步释放特性。由于 HA 修饰，载体对过表达三阴性乳腺癌细胞 MDA-MB-231 的 CD44 具有强大的正靶向性。细胞毒性和凋亡研究证实了两种药物的靶向作用和协同作用。有趣的是，我们在细胞周期研究中发现，药物组合（游离组合或 Combo NCs）并没有显示出 G2M 期的升高，而单独用 DTX 处理时，G2M 期的升高显著更高。我们进一步发现，组合中的 DTX 可能通过调节 GEM 相关酶来发挥作用，从而增强 GEM 的功效。因此，该纳米平台为癌症治疗中实现靶向共递药和增效作用提供了新的解决方案。
