Chitosan Coated Polylactic Acid Nanoparticle-Mediated Combinatorial Delivery of Cisplatin and siRNA/Plasmid DNA Chemosensitizes Cisplatin-Resistant Human Ovarian Cancer Cells
Chitosan Coated Polylactic Acid Nanoparticle-Mediated Combinatorial Delivery of Cisplatin and siRNA/Plasmid DNA Chemosensitizes Cisplatin-Resistant Human Ovarian Cancer Cells
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DOI：
10.1021/mp500259e
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发表时间：
2014-08-01
影响因子：
4.9
通讯作者：
Ramesh, Rajagopal
中科院分区：
文献类型：
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作者：
Babu, Anish;Wang, Qi;Ramesh, Rajagopal
对抗癌药物的耐药性的发展导致治疗无效，导致死亡率增加。因此，克服耐药性和恢复对抗癌药物的敏感性将提高治疗效果和降低死亡率。虽然之前已经证实了癌症耐药的许多机制，但最近的研究暗示蛋白酶体和自噬调节蛋白P62/SQSTM1 （P62）在促进耐药方面的作用。具体而言，已知蛋白酶体β 5亚基表达的减少和/或P62蛋白表达的增强有助于卵巢癌细胞中顺铂（CDDP）等癌症耐药。因此，我们假设在CDDP耐药的卵巢癌细胞中恢复)65的表达和/或抑制P62蛋白的表达将导致对CDDP敏感性的恢复和细胞杀伤的增强。为了验证我们的假设，我们开发了一种可生物降解的多功能纳米颗粒（MNP）系统，该系统可共同递送P62siRNA、β 5质粒DNA和CDDP，并测试了其对CDDP耐药的2008/C13卵巢癌细胞的疗效。MNP由负载CDDP的聚乳酸纳米颗粒作为内核，由离子连接的P62siRNA （siP62）和/或/35表达质粒DNA （p β 5）组成的阳离子壳聚糖（CS）作为外层。MNPs为球形，水动力直径在280 ~ 350 nm之间，对CDDP和siRNA的包封效率分别为8296%和78.5%。通过凝胶阻滞和分光光度法测定，MNPs有效地保护了siRNA，与裸siRNA相比，MNPs表现出更好的血清稳定性。MNPs成功递送了siP62和pfi5，导致P62敲低并恢复2008/C13细胞中/35的表达。交付siP62相结合,使用基于pβ5,CDDP导致明显减少的IC50值2008年CDDP / C13细胞125 + / - 1.3μM 98 + / - 0.6μM (p < 0.05, 21.696)相比,减少的减少ICso CDDP中观察到的细胞只有siP62交付(IC50 = 106 + / - 1.1μM, p < 0.05;减少15.2%)或pβS交付(IC50 = 115 + / - 2.8μM;减少8%)通过基于。最后，我们的研究表明，2008/ c13细胞的CDDP耐药指数从游离CDDP的4.62降低到MNP处理的3.62。总之，我们的研究结果证明了我们的MNP在克服卵巢癌细胞CDDP耐药方面的有效性。
Development of resistance toward anticancer drugs results in ineffective therapy leading to increased mortality. Therefore, overriding resistance and restoring sensitivity to anticancer drugs will improve treatment efficacy and reduce mortality. While numerous mechanisms for drug resistance in cancer have previously been demonstrated, recent studies implicate a role for proteasome and the autophagy regulatory protein P62/SQSTM1 (P62) in contributing to drug resistance. Specifically, reduction in the expression of the beta 5 subunit of the proteasome and/or enhanced P62 protein expression is known to contribute to cancer drug resistance such as cisplatin (CDDP) in ovarian cancer cells. Therefore, we hypothesized that restoration of)65 expression and/or suppression of P62 protein expression in CDDP-resistant ovarian cancer cells will lead to restoration of sensitivity to CDDP and enhanced cell killing. To test our hypothesis we developed a biodegradable multifunctional nanoparticle (MNP) system that codelivered P62siRNA, beta 5 plasmid DNA, and CDDP and tested its efficacy in CDDP resistant 2008/C13 ovarian cancer cells. MNP consisted of CDDP loaded polylactic acid nanoparticle as inner core and cationic chitosan (CS) consisting of ionically linked P62siRNA (siP62) and/or /35 expressing plasmid DNA (p beta 5) as the outer layer. The MNPs were spherical in shape with a hydrodynamic diameter in the range of 280-350 nm, and demonstrated encapsulation efficiencies of 8296 and 78.5% for CDDP and siRNA respectively. MNPs efficiently protected the siRNA and showed superior serum stability compared to naked siRNA as measured by gel retardation and spectrophotometry assays. The MNPs successfully delivered siP62 and pfi5 to cause P62 knockdown and restoration of /35 expression in 2008/C13 cells. Combined delivery of siP62, p beta 5, and CDDP using the MNPs resulted in a marked reduction in the IC50 value of CDDP in 2008/C13 cells from 125 +/- 1.3 mu M to 98 +/- 0.6 mu M (P < 0.05; 21.696 reduction) when compared to the reduction in the ICso of CDDP observed in cells that had only siP62 delivered (IC50 = 106 +/- 1.1 mu M; P < 0.05; 15.2% reduction) or p beta S delivered (IC50 = 115 +/- 2.8 mu M; 8% reduction) via MNPs. Finally, our studies showed that the CDDP resistance index in 2008/C 13 cells was reduced from 4.62 for free CDDP to 3.62 for MNP treatment. In conclusion our study results demonstrated the efficacy of our MNP in overcoming CDDP resistance in ovarian cancer cells.