Skip to main content
SpringerLink
Log in

Vaginal Suppositories with siRNA and Paclitaxel-Incorporated Solid Lipid Nanoparticles for Cervical Cancer: Preparation and In Vitro Evaluation

  • Protocol
  • First Online:
RNA Interference and Cancer Therapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1974))

Abstract

The objective of this study is to prepare vaginal suppository containing chemotherapeutic agent and genetic material that can be applied locally for cervical cancer. Cervical cancer is one of the most life-threatening types of cancer among women and is generally resistant to chemotherapy. Paclitaxel has been selected as chemotherapeutic agent, and siRNA that inhibits the Bcl-2 oncogene has been selected as the genetic material for simultaneous vaginal delivery. For this purpose, three different solid lipid nanoparticles (SLNs) were prepared that include Bcl-2 siRNA and paclitaxel and paclitaxel/Bcl-2 siRNA combination separately, and these SLN formulations were dispersed in vaginal suppositories prepared with PEG 6000. First, the physicochemical properties of SLNs, their cytotoxicities on HeLa cell lines, and the transfection ability of siRNA-incorporated SLN on the cells have been examined. Afterward, the release of SLNs from the three different vaginal suppositories prepared has been determined via horizontal diffusion chamber system. The loaded amount to the SLNs and release amount from suppositories of paclitaxel have been determined via HPLC, whereas stability, loading, and release amount of siRNA has been determined via gel retardation system and UV spectrophotometer.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Urasa M, Darj E (2011) Knowledge of cervical cancer and screening practices of nurses at a regional hospital in Tanzania. Afr Health Sci 11(1):48–57

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Volodko V, Gordon M, Salla M, Ghazaleh HA, Baksh S (2014) RASSF tumor suppressor gene family: biological functions and regulation. FEBS Lett 588:2671–2684

    Article  CAS  PubMed  Google Scholar 

  3. Zhang S, Zhao B, Jiang H, Wang B, Ma B (2007) Cationic lipids and polymers mediated vectors for delivery of siRNA. J Control Release 123:1–10

    Article  CAS  PubMed  Google Scholar 

  4. Xu C, Wang J (2015) Delivery systems for siRNA drug development in cancer therapy. Asian J Pharm Sci 10:1–12

    Article  Google Scholar 

  5. Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411:494–498

    Article  CAS  PubMed  Google Scholar 

  6. Tseng YC, Mozumdar S, Huang L (2009) Lipid-based systemic delivery of siRNA. Adv Drug Deliv Rev 61:721–731

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Li CX, Parker A, Menocal E, Xiang S, Borodyansky L, Fruehauf JH (2006) Delivery of RNA interference. Cell Cycle 5(18):2103–2109

    Article  CAS  PubMed  Google Scholar 

  8. Takabatake Y, Isaka Y, Mizui M, Kawachi H, Takahara S, Imai E (2007) Chemically modified siRNA prolonged RNA interference in renal disease. Biochem Biophys Res Commun 363(2):432–437

    Article  CAS  PubMed  Google Scholar 

  9. Gary D, Puri N, Won YY (2007) Polymer-based siRNA delivery: perspectives on the fundamental and phenomenological distinctions from polymer-based DNA delivery. J Control Release 121:64–73

    Article  CAS  PubMed  Google Scholar 

  10. Reischl D, Zimmer A (2009) Drug delivery of siRNA therapeutics: potentials and limits of nanosystems. Nanomed Nanotechnol Biol Med 5(1):8–20

    Article  CAS  Google Scholar 

  11. Gorbatyuk M, Justilien V, Liu J, Hauswirth WW, Lewin AS (2007) Suppression of mouse rhodopsin expression in vivo by AAV mediated siRNA delivery. VisionRes 47(9):1202–1208

    CAS  Google Scholar 

  12. Chen GX, Dong QH, Zhang JW, Fu FF, Xu ZF, Liang QY, Zheng S, Ding JY (2006) Knockdown of PLK1 mRNA by special siRNA. Yi Chuan 28(1):21–25

    PubMed  Google Scholar 

  13. Sioud M, Sørensen DR (2003) Cationic liposome-mediated delivery of siRNAs in adult mice. Biochem Biophys Res Commun 312(4):1220–1225

    Article  CAS  PubMed  Google Scholar 

  14. Bouxsein NF, Mcallister CS, Ewert KK, Samuel CE, Safinya RC (2007) Structure and gene silencing activities of monovalent and pentavalent cationic lipid vectors complexed with siRNA. Biochemistry 46(16):4785–4792

    Article  CAS  PubMed  Google Scholar 

  15. Kataoka K, Itaka K, Nishiyama N, Yamasaki Y, Oishi M, Nagasaki Y (2005) Smart polymeric micelles as nanocarriers for oligonucleotides and siRNA delivery. Nucleic Acids Symp Ser 49(1):17–18

    Article  Google Scholar 

  16. Katas H, Alpar HO (2006) Development and characterisation of chitosan nanoparticles for siRNA delivery. J Control Release 115(2):16–25

    Article  Google Scholar 

  17. Savaese A, Cognetti F (2003) New drugs in the treatment of recurrent or metastatic cervical cancer. Crit Rev Oncol Hematol 48:323–327

    Article  Google Scholar 

  18. Wu J, Liu Q, Lee RJ (2006) A folate receptor-targeted liposomal formulation for paclitaxel. Int J Pharm 316:148–153

    Article  CAS  PubMed  Google Scholar 

  19. Thu TH, Dass CR, Dunstan DE (2008) Injectable chitosan hydrogels for localized cancer therapy. J Control Release 126:205–216

    Article  Google Scholar 

  20. Chimmiri P, Rajalakshmi R, Mahitha B, Ramesh G, Noor Ahmed VH (2012) Solid lipid nanoparticles: a novel carrier for cancer therapy. Int J Pharm Biol Sci 3(3):405–413

    Google Scholar 

  21. Serpea L, Catalano MG, Cavalli R, Ugazio E, Bosco O, Canaparo R, Muntoni E, Frairia R, Gasco MR, Eandi M, Zara GP (2004) Cytotoxicity of anticancer drugs incorporated in solid lipid nanoparticles on HT-29 colorectal cancer cell line. Eur J Pharm Biopharm 58:673–680

    Article  Google Scholar 

  22. Shah R, Eldridge D, Palombo E, Harding I (2014) Optimisation and stability assessment of solid lipid nanoparticles using particle size and zeta potential. J Phys Sci 25(1):59–75

    CAS  Google Scholar 

  23. Banerjee I, De K, Mukherjee D, Dey D, Chattopadhyay S, Mukherjee M, Mandal M, Bandyopadhyay AK, Gupta A, Ganguly S, Misra M (2016) Paclitaxel-loaded solid lipid nanoparticles modified with Tyr-3-octreotide for enhanced antiangiogenic and anti-glioma therapy. Acta Biomater 38:69–81

    Article  CAS  PubMed  Google Scholar 

  24. Yu YH, Kim E, Park DE, Shim G, Lee S, Kim YB, Kim CW, Oha YK (2012) Cationic solid lipid nanoparticles for co-delivery of paclitaxel and siRNA. Eur J Pharm Biopharm 80:268–273

    Article  CAS  PubMed  Google Scholar 

  25. Pavelić Z, Skalko-Basnet N, Filipović-Grcić J, Martinac A, Jalsenjak I (2005) Development and in vitro evaluation of a liposomal vaginal delivery system for acyclovir. J Control Release 106:34–43

    Article  PubMed  Google Scholar 

  26. Hussain A, Ahsan F (2005) The vagina as a route for systemic drug delivery. J Control Release 103:301–313

    Article  CAS  PubMed  Google Scholar 

  27. Dobaria N, Mashru R, Vadia NH (2007) Vaginal drug delivery systems: a review of current status. East Cent Afr J Pharm Sci 10:3–13

    Google Scholar 

  28. Choudhury A, Das S, Kar M (2011) A review on novelty and potentiality of vaginal drug delivery. Int J PharmTech Res 3(2):1033–1044

    CAS  Google Scholar 

  29. Haas S, Woerdenbag H, Sznitowska M (2015) Rectal and vaginal. In: Bouwman-Boer Y, Fenton-May V, Brun PL (eds) Practical pharmaceutics: an international guideline for the preparation, care and use of medicinal products. Springer International Publishing AG, Cham, pp 190–225

    Google Scholar 

  30. Büyükköroğlu G, Şenel B, Başaran E, Yenilmez E, Yazan Y (2016) Preparation and in vitro evaluation of vaginal formulations including siRNA and paclitaxel-loaded SLNs for cervical cancer. Eur J Pharm Biopharm 109:174–183

    Article  PubMed  Google Scholar 

  31. Büyükköroğlu G, Yazan EY, Öner AF (2015) Preparation and physicochemical characterizations of solid lipid nanoparticles containing DOTAP® for DNA delivery. Turk J Chem 39:1012–1024

    Article  Google Scholar 

  32. Gvili K, Benny O, Danino D, Machluf M (2006) Poly(D,L-lactide-coglycolide acid) nanoparticles for DNA delivery: waiving preparation complexity and increasing efficiency. Biopolymers 85:379–391

    Google Scholar 

  33. Mehnert W, Mäder K (2001) Solid lipid nanoparticles: production, characterization and applications. Adv Drug Deliv Rev 47(2–3):165–196

    Article  CAS  PubMed  Google Scholar 

  34. Şenel B, Büyükköroğlu G, Yazan Y (2015) Solid lipid and chitosan particulate systems for delivery of siRNA. Pharmazie 8:698–705

    Google Scholar 

  35. Neves J, Amaral MH, Bahia MF (2008) Performance of an in vitro mucoadhesion testing method for vaginal semisolids: influence of different testing conditions and instrumental parameters. Eur J Pharm Biopharm 69:622–632

    Article  PubMed  Google Scholar 

  36. Owen DH, Katz DF (1999) A vaginal fluid simulant. Contraception 59:91–95

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Anadolu University, Eskisehir, Turkey

    Gülay Büyükköroğlu & Behiye Şenel

  2. Department of Pharmaceutical Technology, Faculty of Pharmacy, Anadolu University, Eskisehir, Turkey

    Evrim Yenilmez

Authors
  1. Gülay Büyükköroğlu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Behiye Şenel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Evrim Yenilmez
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gülay Büyükköroğlu .

Editor information

Editors and Affiliations

  1. Cancer Biology, CSIR-Centre for Cellular and Molecular Biology, Hyderabad, Telangana, India

    Lekha Dinesh Kumar

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Büyükköroğlu, G., Şenel, B., Yenilmez, E. (2019). Vaginal Suppositories with siRNA and Paclitaxel-Incorporated Solid Lipid Nanoparticles for Cervical Cancer: Preparation and In Vitro Evaluation. In: Dinesh Kumar, L. (eds) RNA Interference and Cancer Therapy. Methods in Molecular Biology, vol 1974. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9220-1_22

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9220-1_22

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9219-5

  • Online ISBN: 978-1-4939-9220-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation