Nano Research

, Volume 10, Issue 9, pp 3049–3067 | Cite as

Biodegradable nanocarriers for small interfering ribonucleic acid (siRNA) co-delivery strategy increase the chemosensitivity of pancreatic cancer cells to gemcitabine

  • Chengbin Yang
  • Kok Ken Chan
  • Wen-Jen Lin
  • Alana Mauluidy Soehartono
  • Guimiao Lin
  • Huiting Toh
  • Ho Sup Yoon
  • Chih-Kuang Chen
  • Ken-Tye Yong
Research Article
  • 98 Downloads

Abstract

Ribonucleic acid (RNA) interference (RNAi) therapies are promising cancer treatment modalities that can specifically target abnormal proto-oncogenes, thus improving the therapeutic effect. For the treatment of pancreatic cancer, targeting one mutant proto-oncogene by RNAi usually does not yield the desired therapeutic efficiency. Both K-ras gene mutations and Notch1 overexpression are common symptoms in pancreatic cancer patients, and play a crucial role in pancreatic cancer cell drug resistance. In this study, biodegradable charged polyester-based vectors (BCPVs) were synthesized for the co-delivery of K-ras and Notch1 small interfering ribonucleic acid (siRNA) into MiaPaCa-2 cells (pancreatic cancer cell line) to overcome drug resistance to gemcitabine (GEM), a first-line chemotherapeutic drug used in the clinic. BCPVs could effectively absorb negative siRNA to form a capsule-like structure, prevent siRNA from nuclease digestion in the serum, and promote effective siRNA cell internalization and endosomal escape. Through K-ras and Notch1 gene silencing in MiaPaCa-2 cells, BCPV-siRNAK-ras-siRNANotch1 nanocomplexes effectively reversed the epithelia-mesenchymal transition (EMT) in MiaPaCa-2 cells, thereby greatly enhancing the sensitivity of MiaPaCa-2 cells to GEM. MiaPaCa-2 cell proliferation, migration, and invasion were effectively inhibited, and cell apoptosis was also significantly enhanced by the synergistic antitumor effect of BCPV-siRNAK-ras-siRNANotch1 nanocomplexes and GEM. These results suggest that this combination RNAi therapy can be used to improve cancer cell sensitivity to chemotherapeutic drugs. Specifically, this newly developed strategy has a great potential for treating pancreatic cancer.

Keywords

pancreatic cancer small interfering ribonucleic acid (siRNA) biodegradable charged polyester-based vector (BCPV) gemcitabine (GEM) epithelia-mesenchymal transition (EMT) 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Supplementary material

12274_2017_1521_MOESM1_ESM.pdf (2.7 mb)
Biodegradable nanocarriers for small interfering ribonucleic acid (siRNA) co-delivery strategy increase the chemosensitivity of pancreatic cancer cells to gemcitabine

References

  1. [1]
    Siegel, R.; Ma, J. M.; Zou, Z. H.; Jemal, A. Cancer statistics, 2014. CACancer J.Clin. 2014, 64, 9–29.CrossRefGoogle Scholar
  2. [2]
    Siegel, R. L.; Miller, K. D.; Jemal, A. Cancer statistics, 2016. CA Cancer J. Clin. 2016, 66, 7–30.CrossRefGoogle Scholar
  3. [3]
    Waddell, N.; Pajic, M.; Patch, A. M.; Chang, D. K.; Kassahn, K. S.; Bailey, P.; Johns, A. L.; Miller, D.; Nones, K.; Quek, K. et al. Whole genomes redefine the mutational landscape of pancreatic cancer. Nature 2015, 518, 495–501.CrossRefGoogle Scholar
  4. [4]
    Fleming, J. B.; Shen, G. L.; Holloway, S. E.; Davis, M.; Brekken, R. A. Molecular consequences of silencing mutant K-ras in pancreatic cancer cells: Justification for K-rasdirected therapy. Mol. Cancer Res. 2005, 3, 413–423.CrossRefGoogle Scholar
  5. [5]
    Yang, C. B.; Hu, R.; Anderson, T.; Wang, Y. C.; Lin, G. M.; Law, W. C.; Lin, W. J.; Nguyen, Q. T.; Toh, H. T.; Yoon, H. S. et al. Biodegradable nanoparticle-mediated K-ras down regulation for pancreatic cancer gene therapy. J. Mater. Chem. B 2015, 3, 2163–2172.CrossRefGoogle Scholar
  6. [6]
    Vakiani, E.; Solit, D. B. KRAS and BRAF: Drug targets and predictive biomarkers. J. Pathol. 2011, 223, 220–230.CrossRefGoogle Scholar
  7. [7]
    Weijzen, S.; Rizzo, P.; Braid, M.; Vaishnav, R.; Jonkheer, S. M.; Zlobin, A.; Osborne, B. A.; Gottipati, S.; Aster, J. C.; Hahn, W. C. et al. Activation of Notch-1 signaling maintains the neoplastic phenotype in human ras-transformed cells. Nat. Med. 2002, 8, 979–986.CrossRefGoogle Scholar
  8. [8]
    Miyamoto, Y.; Maitra, A.; Ghosh, B.; Zechner, U.; Argani, P.; Iacobuzio-Donahue, C. A.; Sriuranpong, V.; Iso, T.; Meszoely, I. M.; Wolfe, M. S. et al. Notch mediates TGFa-induced changes in epithelial differentiation during pancreatic tumorigenesis. Cancer Cell 2003, 3, 565–576.CrossRefGoogle Scholar
  9. [9]
    Bao, B.; Wang, Z. W.; Ali, S.; Kong, D. J.; Li, Y. W.; Ahmad, A.; Banerjee, S.; Azmi, A. S.; Miele, L.; Sarkar, F. H. Notch-1 induces epithelial–mesenchymal transition consistent with cancer stem cell phenotype in pancreatic cancer cells. Cancer Lett. 2011, 307, 26–36.CrossRefGoogle Scholar
  10. [10]
    Avila, J. L.; Kissil, J. L. Notch signaling in pancreatic cancer: Oncogene or tumor suppressor? Trends Mol. Med. 2013, 19, 320–327.Google Scholar
  11. [11]
    De La J.P.; Emerson, L. L.; Goodman, J. L.; Froebe, S. C.; Illum, B. E.; Curtis, A. B.; Murtaugh, L. C. Notch and Kras reprogram pancreatic acinar cells to ductal intraepithelial neoplasia. Proc. Natl. Acad. Sci. USA 2008, 105, 18907–18912.CrossRefGoogle Scholar
  12. [12]
    Yabuuchi, S.; Pai, S. G.; Campbell, N. R.; de Wilde, R. F.; De Oliveira, E.; Korangath, P.; Streppel, M. M.; Rasheed, Z. A.; Hidalgo, M.; Maitra, A. et al. Notch signaling pathway targeted therapy suppresses tumor progression and metastatic spread in pancreatic cancer. Cancer Lett. 2013, 335, 41–51.CrossRefGoogle Scholar
  13. [13]
    Wang, Z. W.; Banerjee, S.; Li, Y. W.; Rahman, K. M. W.; Zhang, Y. X.; Sarkar, F. H. Down-regulation of Notch-1 inhibits invasion by inactivation of nuclear factor-κB, vascular endothelial growth factor, and matrix metalloproteinase-9 in pancreatic cancer cells. Cancer Res. 2006, 66, 2778–2784.CrossRefGoogle Scholar
  14. [14]
    Ji, Z. Y.; Mei, F. C.; Xie, J. W.; Cheng, X. D. Oncogenic KRAS activates hedgehog signaling pathway in pancreatic cancer cells. J. Biol. Chem. 2007, 282, 14048–14055.CrossRefGoogle Scholar
  15. [15]
    Collins, M. A.; Brisset, J. C.; Zhang, Y. Q.; Bednar, F.; Pierre, J.; Heist, K. A.; Galbán, C. J.; Galbán, S.; di Magliano, M. P. Metastatic pancreatic cancer is dependent on oncogenic Kras in mice. PLoS One 2012, 7, e49707.CrossRefGoogle Scholar
  16. [16]
    Burris, H. A.; Moore, M. J.; Andersen, J.; Green, M. R.; Rothenberg, M. L.; Modiano, M. R.; Cripps, M. C.; Portenoy, R. K.; Storniolo, A. M.; Tarassoff, P. et al. Improvements in survival and clinical benefit with gemcitabine as first-line therapy for patients with advanced pancreas cancer: Arandomized trial. J. Clin. Oncol. 1997, 15, 2403–2413.CrossRefGoogle Scholar
  17. [17]
    Conroy, T.; Desseigne, F.; Ychou, M.; Bouché, O.; Guimbaud, R.; Bécouarn, Y.; Adenis, A.; Raoul, J. L.; Gourgou-Bourgade, S.; de la Fouchardière, C. et al. FOLFIRINOX versus gemcitabine for metastatic pancreatic cancer. N. Engl. J. Med. 2011, 364, 1817–1825.Google Scholar
  18. [18]
    Mittal, A.; Chitkara, D.; Behrman, S. W.; Mahato, R. I. Efficacy of gemcitabine conjugated and miRNA-205 complexed micelles for treatment of advanced pancreatic cancer. Biomaterials 2014, 35, 7077–7087.CrossRefGoogle Scholar
  19. [19]
    Lima, C. M. S. R.; Savarese, D.; Bruckner, H.; Dudek, A.; Eckardt, J.; Hainsworth, J.; Yunus, F.; Lester, E.; Miller, W.; Saville, W. et al. Irinotecan plus gemcitabine induces both radiographic and CA19–9 tumor marker responses in patients with previously untreated advanced pancreatic cancer. J. Clin. Oncol. 2002, 20, 1182–1191.CrossRefGoogle Scholar
  20. [20]
    Berlin, J. D.; Adak, S.; Vaughn, D. J.; Flinker, D.; Blaszkowsky, L.; Harris, J. E.; Al Benson, B. III. A phase II study of gemcitabine and 5-fluorouracil in metastatic pancreatic cancer: An eastern cooperative oncology group study (E3296). Oncology 2000, 58, 215–218.CrossRefGoogle Scholar
  21. [21]
    Lin, G. M.; Hu, R.; Law, W. C.; Chen, C. K.; Wang, Y. C.; Li, C. H.; Nguyen, Q. T.; Lai, C. K.; Yoon, H. S.; Wang, X. M. et al. Biodegradable nanocapsules as siRNA carriers for mutant K-ras gene silencing of human pancreatic carcinoma cells. Small 2013, 9, 2757–2763.CrossRefGoogle Scholar
  22. [22]
    Lin, G. M.; Yang, C. B.; Hu, R.; Chen, C. K.; Law, W. C.; Anderson, T.; Zhang, B. T.; Nguyen, Q. T.; Toh, H. T.; Yoon, H. S. et al. Interleukin-8 gene silencing on pancreatic cancer cells using biodegradable polymer nanoplexes. Biomater. Sci. 2014, 2, 1007–1015.CrossRefGoogle Scholar
  23. [23]
    Zheng, N.; Song, Z. Y.; Liu, Y.; Zhang, R. J.; Zhang, R. Y.; Yao, C.; Uckun, F. M.; Yin, L. C.; Cheng, J. J. Redoxresponsive, reversibly-crosslinked thiolated cationic helical polypeptides for efficient siRNA encapsulation and delivery. J. Control. Release 2015, 205, 231–239.CrossRefGoogle Scholar
  24. [24]
    Wang, Z. W.; Li, Y. W.; Kong, D. J.; Banerjee, S.; Ahmad, A.; Azmi, A. S.; Ali, S.; Abbruzzese, J. L.; Gallick, G. E.; Sarkar, F. H. Acquisition of epithelial–mesenchymal transition phenotype of gemcitabine-resistant pancreatic cancer cells is linked with activation of the notch signaling pathway. Cancer Res. 2009, 69, 2400–2407.CrossRefGoogle Scholar
  25. [25]
    Liu, C. X.; Zhao, G.; Liu, J.; Ma, N. C.; Chivukula, P.; Perelman, L.; Okada, K.; Chen, Z. Y.; Gough, D.; Yu, L. Novel biodegradable lipid nano complex for siRNA delivery significantly improving the chemosensitivity of human colon cancer stem cells to paclitaxel. J.Control. Release 2009, 140, 277–283.CrossRefGoogle Scholar
  26. [26]
    Birmingham, A.; Anderson, E.; Sullivan, K.; Reynolds, A.; Boese, Q.; Leake, D.; Karpilow, J.; Khvorova, A. A protocol for designing siRNAs with high functionality and specificity. Nat. Protoc. 2007, 2, 2068–2078.CrossRefGoogle Scholar
  27. [27]
    Conde, J.; Ambrosone, A.; Hernandez, Y.; Tian, F. R.; McCully, M.; Berry, C. C.; Baptista, P. V.; Tortiglione, C.; de la Fuente, J. M. 15 years on siRNA delivery: Beyond the state-of-the-art on inorganic nanoparticles for RNAi therapeutics. Nano Today 2015, 10, 421–450.CrossRefGoogle Scholar
  28. [28]
    Wang, J.; Lu, Z.; Wientjes, M. G.; Au, J. L. S. Delivery of siRNA therapeutics: Barriers and carriers. AAPS J. 2010, 12, 492–503.CrossRefGoogle Scholar
  29. [29]
    Kanasty, R.; Dorkin, J. R.; Vegas, A.; Anderson, D. Delivery materials for siRNA therapeutics. Nat. Mater. 2013, 12, 967–977.CrossRefGoogle Scholar
  30. [30]
    Zhao, X.; Li, F.; Li, Y. Y.; Wang, H.; Ren, H.; Chen, J.; Nie, G. J.; Hao, J. H. Co-delivery of HIF1a siRNA and gemcitabine via biocompatible lipid-polymer hybrid nanoparticles for effective treatment of pancreatic cancer. Biomaterials 2015, 46, 13–25.CrossRefGoogle Scholar
  31. [31]
    Ozcan, G.; Ozpolat, B.; Coleman, R. L.; Sood, A. K.; Lopez-Berestein, G. Preclinical and clinical development of siRNA-based therapeutics. Adv. Drug Deliv. Rev. 2015, 87, 108–119.CrossRefGoogle Scholar
  32. [32]
    Davidson, B. L.; McCray, P. B. Current prospects for RNA interference-based therapies. Nat. Rev. Gen. 2011, 12, 329–340.CrossRefGoogle Scholar
  33. [33]
    Petrocca, F.; Lieberman, J. Promise and challenge of RNA interference-based therapy for cancer. J. Clin. Oncol. 2011, 29, 747–754.CrossRefGoogle Scholar
  34. [34]
    Yang, C. B.; Panwar, N.; Wang, Y. C.; Zhang, B. T.; Liu, M. X.; Toh, H.; Yoon, H. S.; Tjin, S. C.; Chong, P. H. J.; Law, W. C. et al. Biodegradable charged polyester-based vectors (BCPVs) as an efficient non-viral transfection nanoagent for gene knockdown of the BCR-ABL hybrid oncogene in a human chronic myeloid leukemia cell line. Nanoscale 2016, 8, 9405–9416.CrossRefGoogle Scholar
  35. [35]
    Jones, C. H.; Chen, C. K.; Jiang, M.; Fang, L.; Cheng, C.; Pfeifer, B. A. Synthesis of cationic polylactides with tunable charge densities as nanocarriers for effective gene delivery. Mol. Pharmaceutics 2013, 10, 1138–1145.CrossRefGoogle Scholar
  36. [36]
    Wang, Y. C.; Wu, B.; Yang, C. B.; Liu, M. X.; Sum, T. C.; Yong, K. T. Synthesis and characterization of Mn: ZnSe/ZnS/ZnMnS sandwiched QDs for multimodal imaging and theranostic applications. Small 2016, 12, 534–546.CrossRefGoogle Scholar
  37. [37]
    Ngamcherdtrakul, W.; Morry, J.; Gu, S. D.; Castro, D. J.; Goodyear, S. M.; Sangvanich, T.; Reda, M. M.; Lee, R.; Mihelic, S. A.; Beckman, B. L. et al. Cationic polymer modified mesoporous silica nanoparticles for targeted siRNA delivery to HER2+ breast cancer. Adv. Funct. Mater. 2015, 25, 2646–2659.CrossRefGoogle Scholar
  38. [38]
    Song, L. L.; Peng, Y.; Yun, J.; Rizzo, P.; Chaturvedi, V.; Weijzen, S.; Kast, W. M.; Stone, P. J. B.; Santos, L.; Loredo, A. et al. Notch-1 associates with IKKa and regulates IKK activity in cervical cancer cells. Oncogene 2008, 27, 5833–5844.CrossRefGoogle Scholar
  39. [39]
    Yang, C.; Mo, X.; Lv, J.; Liu, X.; Yuan, M.; Dong, M.; Li, L.; Luo, X.; Fan, X.; Jin, Z. Lipopolysaccharide enhances FcεRI-mediated mast cell degranulation by increasing Ca2+ entry through store-operated Ca2+ channels: Implications for lipopolysaccharide exacerbating allergic asthma. Exp. Physiol. 2012, 97, 1315–1327.CrossRefGoogle Scholar
  40. [40]
    Panwar, N.; Yang, C. B.; Yin, F.; Yoon, H. S.; Chuan, T. S.; Yong, K. T. RNAi-based therapeutic nanostrategy: IL-8 gene silencing in pancreatic cancer cells using gold nanorods delivery vehicles. Nanotechnology 2015, 26, 365101.CrossRefGoogle Scholar
  41. [41]
    Yin, F.; Yang, C. B.; Wang, Q. Q.; Zeng, S. W.; Hu, R.; Lin, G. M.; Tian, J. L.; Hu, S. Y.; Lan, R. F.; Yoon, H. S. et al. A light-driven therapy of pancreatic adenocarcinoma using gold nanorods-based nanocarriers for co-delivery of doxorubicin and siRNA. Theranostics 2015, 5, 818–833.CrossRefGoogle Scholar
  42. [42]
    Lennon, A. M.; Wolfgang, C. L.; Canto, M. I.; Klein, A. P.; Herman, J. M.; Goggins, M.; Fishman, E. K.; Kamel, I.; Weiss, M. J.; Diaz, L. A. et al. The early detection of pancreatic cancer: What will it take to diagnose and treat curable pancreatic neoplasia? Cancer Res. 2014, 74, 3381–3389.CrossRefGoogle Scholar
  43. [43]
    Anderson, T.; Hu, R.; Yang, C. B.; Yoon, H. S.; Yong, K. T. Pancreatic cancer gene therapy using an siRNA-functionalized single walled carbon nanotubes (SWNTs) nanoplex. Biomater. Sci. 2014, 2, 1244–1253.CrossRefGoogle Scholar
  44. [44]
    Hu, R.; Yang, C. B.; Wang, Y. C.; Lin, G. M.; Qin, W.; Ouyan, Q. L.; Law, W. C.; Nguyen, Q. T.; Yoon, H. S.; Wang, X. M. et al. Aggregation-induced emission (AIE) dye loaded polymer nanoparticles for gene silencing in pancreatic cancer and their in vitro and in vivo biocompatibility evaluation. Nano Res. 2015, 8, 1563–1576.CrossRefGoogle Scholar
  45. [45]
    Li, L.; Gu, W. Y.; Liu, J.; Yan, S. Y.; Xu, Z. P. Aminefunctionalized SiO2 nanodot-coated layered double hydroxide nanocomposites for enhanced gene delivery. Nano Res. 2015, 8, 682–694.CrossRefGoogle Scholar
  46. [46]
    McCully, M.; Hernandez, Y.; Conde, J.; Baptista, P. V.; de la Fuente, J. M.; Hursthouse, A.; Stirling, D.; Berry, C. C. Significance of the balance between intracellular glutathione and polyethylene glycol for successful release of small interfering RNA from gold nanoparticles. Nano Res. 2015, 8, 3281–3292.CrossRefGoogle Scholar
  47. [47]
    Aied, A.; Greiser, U.; Pandit, A.; Wang, W. X. Polymer gene delivery: Overcoming the obstacles. Drug Discov. Today 2013, 18, 1090–1098.CrossRefGoogle Scholar
  48. [48]
    De La O, J.P.; Murtaugh, L. C. Notch and Kras in pancreatic cancer: At the crossroads of mutation, differentiation and signaling. Cell Cycle 2009, 8, 1860–1864.CrossRefGoogle Scholar
  49. [49]
    Sundaram, M. V. The love-hate relationship between Ras and Notch. Genes Dev. 2005, 19, 1825–1839.CrossRefGoogle Scholar
  50. [50]
    Ali, S.; Ahmad, A.; Banerjee, S.; Padhye, S.; Dominiak, K.; Schaffert, J. M.; Wang, Z. W.; Philip, P. A.; Sarkar, F. H. Gemcitabine sensitivity can be induced in pancreatic cancer cells through modulation of miR-200 and miR-21 expression by curcumin or its analogue CDF. Cancer Res. 2010, 70, 3606–3617.CrossRefGoogle Scholar
  51. [51]
    Cheng, G. Z.; Chan, J.; Wang, Q.; Zhang, W. Z.; Sun, C. D.; Wang, L. H. Twist transcriptionally up-regulates AKT2 in breast cancer cells leading to increased migration, invasion, and resistance to paclitaxel. Cancer Res. 2007, 67, 1979–1987.CrossRefGoogle Scholar
  52. [52]
    Fuchs, B. C.; Fujii, T.; Dorfman, J. D.; Goodwin, J. M.; Zhu, A. X.; Lanuti, M.; Tanabe, K. K. Epithelial-tomesenchymal transition and integrin-linked kinase mediate sensitivity to epidermal growth factor receptor inhibition in human hepatoma cells. Cancer Res. 2008, 68, 2391–2399.CrossRefGoogle Scholar
  53. [53]
    Thiery, J. P. Epithelial–mesenchymal transitions in tumour progression. Nat. Rev. Cancer 2002, 2, 442–454.CrossRefGoogle Scholar
  54. [54]
    Brabletz, T.; Jung, A.; Reu, S.; Porzner, M.; Hlubek, F.; Kunz-Schughart, L. A.; Knuechel, R.; Kirchner, T. Variable ß-catenin expression in colorectal cancers indicates tumor progression driven by the tumor environment. Proc. Natl. Acad. Sci. USA 2001, 98, 10356–10361.CrossRefGoogle Scholar
  55. [55]
    Li, Y. W.; VandenBoom, T. G.; Kong, D. J.; Wang, Z. W.; Ali, S.; Philip, P. A.; Sarkar, F. H. Up-regulation of miR-200 and let-7 by natural agents leads to the reversal of epithelial-to-mesenchymal transition in gemcitabine-resistant pancreatic cancer cells. Cancer Res. 2009, 69, 6704–6712.CrossRefGoogle Scholar

Copyright information

© Tsinghua University Press and Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Chengbin Yang
    • 1
  • Kok Ken Chan
    • 1
  • Wen-Jen Lin
    • 2
  • Alana Mauluidy Soehartono
    • 1
  • Guimiao Lin
    • 3
  • Huiting Toh
    • 4
  • Ho Sup Yoon
    • 4
    • 5
  • Chih-Kuang Chen
    • 2
  • Ken-Tye Yong
    • 1
  1. 1.School of Electrical and Electronic EngineeringNanyang Technological UniversitySingaporeSingapore
  2. 2.Polymeric Biomaterials Lab, Department of Fiber and Composite MaterialsFeng Chia UniversityTaichungTaiwan
  3. 3.Key Lab of Biomedical Engineering, School of MedicineShenzhen UniversityShenzhenChina
  4. 4.Division of Structural Biology & Biochemistry, School of Biological SciencesNanyang Technological UniversitySingaporeSingapore
  5. 5.Department of Genetic Engineering, College of Life SciencesKyung Hee UniversityYongin-si Gyeonggi-doRepublic of Korea

Personalised recommendations