Cytological Study in vitro on Co-delivery of siRNA and Paclitaxel within Solid Lipid Nanoparticles to Overcome Multidrug Resistance in Tumors_Journal of Biomedical Engineering

Authors：

HUANGRui ^1,2,3 , YAOXinyu ⁴ , CHENYuan ^1,2,3 , SUNXun ⁴ ,  LINYunzhu ^1,2,3

1. Department of Pharmacy, West China Second University Hospital, Sichuan University, Chengdu 610041, China;
2. Evidence-Based Pharmacy Center, West China Second University Hospital, Sichuan University, Chengdu, 610041, China;
3. Key Laboratory of Birth Defects and Related Diseases of Women and Children ,Sichuan University, Ministry of Education, Chengdu 610041, China;
4. Key Laboratory of Drug Targeting and Drug Delivery Systems, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu 610041, China;

Corresponding author：

LINYunzhu, Email: linyunzhu99@163.com

Keywords：

small interfering RNA; paclitaxel; solid lipid nanoparticles; multidrug resistance; P-glycoprotein

DOI：

10.7507/1001-5515.20160020

Video：

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Multidrug resistance (MDR) remains the major obstacle to the success of clinical cancer chemotherapy. P-glycoprotein (P-gp), encoded by the MDR1, is an important part with complex mechanisms associated with the MDR. In order to overcome the MDR of tumors, we in the present experimental design incorporated small interfering RNA (siRNA) targeting MDR1 gene and anticancer drug paclitaxel (PTX) into the solid lipid nanoparticles (SLNs) to achieve the combinational therapeutic effects of genetherapy and chemotherapy. In this study, siRNA-PTX-SLNs were successfully prepared. The cytotoxicity of blank SLNs and siRNA-PTX-SLNs in MCF-7 cells and MCF-7/ADR cells were detected by MTT; and the uptake efficiency of PTX in MCF-7/ADR cells were detected via HPLC method; quantitative real-time PCR and flow cytometry were performed to investigate the silencing effect of siRNA-PTX-SLNs on MDR1 gene in MCF-7/ADR cells. The results showed that PTX loaded SLNs could significantly inhibit the growth of tumor cells, and more importantly, the MDR tumor cells treated with siRNA-PTX-SLNs showed the lowest viability. HPLC study showed that SLNs could enhance the cellular uptake for PTX. Meanwhile, siRNA delivered by SLNs significantly decreased the P-gp expression in MDR tumor cells, thus increased the cellular accumulation of rhodamine123 as a P-gp substrate. In conclusion, the MDR1 gene could be silenced by siRNA-PTX-SLNs, which could promote the growth inhibition efficiency of PTX on tumor cells, leading to synergetic effect on MDR tumor therapy.

Citation： HUANGRui, YAOXinyu, CHENYuan, SUNXun, LINYunzhu. Cytological Study in vitro on Co-delivery of siRNA and Paclitaxel within Solid Lipid Nanoparticles to Overcome Multidrug Resistance in Tumors. Journal of Biomedical Engineering, 2016, 33(1): 108-114. doi: 10.7507/1001-5515.20160020 Copy

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20.	CARRSTENSEN H, MLLER R H, MLLER B W. Particle size, surface hydrophobicity and interaction with serum of parenteral fat emulsions and model drug carriers as parameters related to RES uptake[J]. Clin Nutr, 1992, 11(5): 289-297.
21.	XIA Zhongsheng, ZHU Zhaohua, ZHANG Liyong, et al. Specific reversal of MDR1/P-gp-dependent multidrug resistance by RNA interference in colon cancer cells[J]. Oncol Rep, 2008, 20(6): 1433-1439.
22.	STEGE A, KRHN A, LAGE H. Overcoming multidrug resistance by RNA interference[J]. Methods Mol Biol, 2010, 596: 447-465.
23.	BUDWORTH J, DAVIES R, MALKHANDI J, et al. Comparison of staurosporine and four analogues: their effects on growth, rhodamine 123 retention and binding to P-glycoprotein in multidrug-resistant MCF-7/Adr cells[J]. Br J Cancer, 1996, 73(9): 1063-1068.
24.	CHEN Liming, LIANG Yongju, RUAN Jiwu, et al. Reversal of P-gp mediated multidrug resistance in-vitro and in-vivo by FG020318[J]. J Pharm Pharmacol, 2004, 56(8): 1061-1066.

1. FUKUMURA D, JAIN R K. Tumor microvasculature and microenvironment: targets for anti-angiogenesis and normalization[J]. Microvasc Res, 2007, 74(2-3): 72-84.
2. CAMPBELL R B. Tumor physiology and delivery of nanopharmaceuticals[J]. Anticancer Agents Med Chem, 2006, 6(6): 503-512.
3. REED J C. Bcl-2: prevention of apoptosis as a mechanism of drug resistance[J]. Hematol Oncol Clin North Am, 1995, 9(2): 451-473.
4. MORROW C S, COWAN K H. Glutathiones-transferases and drug resistance[J]. Cancer Cells, 1990, 2(1): 15-22.
5. DONG Xiaowei, MUMPER R J. Nanomedicinal strategies to treat multidrug-resistant tumors: current progress[J]. Nanomedicine, 2010, 5(4): 597-615.
6. 许景峰．肿瘤细胞MDR1基因多药耐药的研究进展[J]．解放军药学学报，2010，26（5）：453-455.
7. CHEN Y, BATHULA S R, LI Jun, et al. Multifunctional nanoparticles delivering small interfering RNA and doxorubicin overcome drug resistance in cancer[J]. J Biol Chem, 2010, 285(29): 22639-22650.
8. TSENG Y C, MOZUMDAR S, HUANG L. Lipid-based systemic delivery of siRNA[J]. Adv Drug Deliv Rev, 2009, 61(9): 721-731.
9. YADAV S, VAN VLERKEN L E, LITTLE S R, et al. Evaluations of combination MDR-1 gene silencing and paclitaxel administration in biodegradable polymeric nanoparticle formulations to overcome multidrug resistance in cancer cells[J]. Cancer Chemother Pharmacol, 2009, 63(4): 711-722.
10. WU H, HAIT W N, YANG J M. Small interfering RNA-induced suppression of MDR1 (P-glycoprotein) restores sensitivity to multidrug-resistant cancer cells[J]. Cancer Res, 2003, 63(7): 1515-1519.
11. 姚欣宇，石三军，孙逊．小分子干扰RNA-紫杉醇固体脂质纳米粒的制备[J]．华西药学杂志，2014，29（4）：368-370.
12. JP2〗LEE M, KOH W S, HAN S S. Down-regulation of Raf-1 kinase is associated with paclitaxel resistance in human breast cancer MCF-7/Adr cells[J]. Cancer Lett, 2003, 193(1): 57-64.JP〗.
13. YAO Hongjuan, JU Ruijun, WANG Xiaoxing, et al. The antitumor efficacy of functional paclitaxel nanomicelles in treating resistant breast cancers by oral delivery[J]. Biomaterials, 2011, 32(12): 3285-3302.
14. LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J]. Methods, 2001, 25(4): 402-408.
15. SAIJO N. Chemotherapy: the more the better? Overview[J]. Cancer Chemother Pharmacol, 1997, 40(Suppl): S100-S106.
16. JP2〗TEODORI E, DEI S, MARTELLI C, et al. The functions and structure of ABC transporters: implications for the design of new inhibitors of Pgp and MRP1 to control multidrug resistance (MDR)[J]. Curr Drug Targets, 2006, 7(7): 893-909.
17. FOJO A T, UEDA K, SLAMON D J, et al. Expression of a multidrug-resistance gene in human tumors and tissues[J]. Proc Natl Acad Sci U S A, 1987, 84(1): 265-269.
18. MEHNERT W, MADER K. Solid lipid nanoparticles-Production, characterization and applications[J]. Adv Drug Deliv Rev, 2001, 47(2/3): 165-196.
19. TORCHILIN V. Tumor delivery of macromolecular drugs based on the EPR effect[J]. Adv Drug Deliv Rev, 2011, 63(3): 131-135.
20. CARRSTENSEN H, MLLER R H, MLLER B W. Particle size, surface hydrophobicity and interaction with serum of parenteral fat emulsions and model drug carriers as parameters related to RES uptake[J]. Clin Nutr, 1992, 11(5): 289-297.
21. XIA Zhongsheng, ZHU Zhaohua, ZHANG Liyong, et al. Specific reversal of MDR1/P-gp-dependent multidrug resistance by RNA interference in colon cancer cells[J]. Oncol Rep, 2008, 20(6): 1433-1439.
22. STEGE A, KRHN A, LAGE H. Overcoming multidrug resistance by RNA interference[J]. Methods Mol Biol, 2010, 596: 447-465.
23. BUDWORTH J, DAVIES R, MALKHANDI J, et al. Comparison of staurosporine and four analogues: their effects on growth, rhodamine 123 retention and binding to P-glycoprotein in multidrug-resistant MCF-7/Adr cells[J]. Br J Cancer, 1996, 73(9): 1063-1068.
24. CHEN Liming, LIANG Yongju, RUAN Jiwu, et al. Reversal of P-gp mediated multidrug resistance in-vitro and in-vivo by FG020318[J]. J Pharm Pharmacol, 2004, 56(8): 1061-1066.

Journal of Biomedical Engineering

Cytological Study in vitro on Co-delivery of siRNA and Paclitaxel within Solid Lipid Nanoparticles to Overcome Multidrug Resistance in Tumors

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