Enhanced in vitro cytotoxicity and intracellular uptake of Genipin via loaded on Nano-Liposomes made from soy lecithin in MCF-7 cells

Document Type : Research Paper


1 Nano Drug Delivery Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran

2 Substance Abuse Prevention Research Center, Health Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran

3 Pharmaceutical Research Center, Health Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran


Objective(s): As an alternative to chemical drugs, natural compounds such as Genipin can reduce toxicity and side effects. In recent years, Genipin's antioxidant properties have been considered a potential cancer treatment. Therefore, the present study investigated anti-cancer activity of newly formulated nano-liposomal loaded Genipin, made from soy lecithin, against MCF-7 cancer cell line.

Materials and Methods: After synthesis, the physicochemical properties of the liposomes were confirmed by Dynamic light scattering (DLS), Scanning Electron Microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), and UV-vis spectrophotometry.

Results: Our results showed that the prepared nano-liposome had a diameter of 166.2 nm. Its Zeta potential was -25.4 mV which indicates the good electrostatic stability of nano-liposomes. Also, a slight size distribution (PDI 0.2870) and a high encapsulation efficiency (EE% >82% and DL>28%) are other features of synthesized nano-liposomal loaded Genipin. The in vitro result profile demonstrated that the drug-controlled release from Genipin loaded-liposomal is 65% during 70h. The in vitro cytotoxic activity of nano-liposomal loaded Genipin in comparison with free Genipin, was explored on MCF-7 cell line using MTT colorimetric assay. Our results revealed that the IC50% (cytotoxicity) of MCF-7 cells treated with nano-liposomes loaded Genipin were higher than those treated with free Genipin (about 2.4 orders of magnitude). Additionally, cell uptake studies evidenced a higher uptake of negative nano-liposomal loaded Genipin.

Conclusion: In a nutshell, newly formulated nano-liposomal is an ideal vehicle for negative targeting (anticancer effect) of drugs to tumor cells that may result in improved efficacy and reduced toxicity of encapsulated drug moiety.


1.    Ko EY, Moon A. Natural products for chemoprevention of breast cancer. J Cancer Prev. 2015;20(4):223. 
2.    Freitas A, Kaplum V, Rossi DCP,  Buffoni R, Carvalho M, Tabordaac P, et al. Proanthocyanidin polymeric tannins from Stryphnodendron adstringens are effective against Candida spp. isolates and for vaginal candidiasis treatment. J. Ethnopharmacol. 2018;216(5):184-190.
3.    Ye J, Xiangfeng J, Wang L. Medicinal supplement genipin induces p53 and Bax‑dependent apoptosis in colon cancer cells. Oncol. Lett. 2018;16(3):2957-2964.
4.    Nam KN, Kyong Choib, Yo Gun, H Parkc, H Sang, Ki et al, Genipin inhibits the inflammatory response of rat brain microglial cells. Int. Immunopharmacol. 2010;10(4):493-499.
5.    Wang R, Yung KC, Zhao YJ, Poon K. A mechanism for the temporal potentiation of genipin to the cytotoxicity of cisplatin in colon cancer cells. Int J Med Sci. 2016;13(7):507.
6.    Kim ES, Jeong CS, Moon A. Genipin, a constituent of Gardenia jasminoides Ellis, induces apoptosis and inhibits invasion in MDA-MB-231 breast cancer cells. Oncol Rep. 2012;27(2):567-572.
7.    Shanmugam MK, Shen H, Tang F, Arfuso F, Rajesh M, Wang L, et al, Potential role of genipin in cancer therapy. Pharmacol Res. 2018;133(5):195-200.
8.    Jo M, Jeong S, Yun HK, Kim DY,  Kim BR, Kim JL, et al, Genipin induces mitochondrial dysfunction and apoptosis via downregulation of Stat3/mcl-1 pathway in gastric cancer. BMC Cancer. 2019;19(1):1-12.
9.    Ayyasamy V, Owens KM, Desouki MM, Liang P, Bakin A, Thangaraj K, et al, Cellular model of Warburg effect identifies tumor promoting function of UCP2 in breast cancer and its suppression by genipin. PLoS One. 2011;6(9):247-92.
10.    Kreiter J, Rupprecht A, Zimmermann L, Moschinger M, Rokitskaya T, Antonenko Y, et al. Molecular mechanisms responsible for pharmacological effects of genipin on mitochondrial proteins. Biophys J. 2019;117(10):1845-1857.
11.    Akbarzadeh A, Rezaei-Sadabady R, Davaran S, Joo S, Zarghami N, Hanifehpour Y, et al. Liposome: classification, preparation, and applications. Nanoscale Res Lett. 2013;8 (1):1-9.
12.    Fakhravar Z, Ebrahimnejad P, Daraee H, Akbarzadeh A,, et al. Nanoliposomes: Synthesis methods and applications in cosmetics. J Cosmet. 2016;18(3):174-181.
13.    Hadian Z, Sahari MA. Moghimi HR, Barzegard M, et al. Formulation, characterization and optimization of liposomes containing eicosapentaenoic and docosahexaenoic acids; a methodology approach. Iran J Pharm Res. 2014;13(2):393.
14.    Madni A, Shah K, Tariq M, Baloch A, Kanwal R, et al. Devising interactive dissolution experiment for pharmacy students (part II): use of dialysis bag method to evaluate effect of dialysis bag length on drug release from novel drug delivery systems. J Biol Res. 2017;2(1):7-12.
15.    Madankan R, Pouget S, Singla P, Bursik M, Dehne J, Jones M, Patra J, Pavolonisf M, et al. Computation of probabilistic hazard maps and source parameter estimation for volcanic ash transport and dispersion. J Comput Phys. 2014;271(5):39-59.
16.    Aktas B, Kasimir-Bauer S, Müller V, Janni W, Fehm T, Wallwiener D, et al. Comparison of the HER2, estrogen and progesterone receptor expression profile of primary tumor, metastases and circulating tumor cells in metastatic breast cancer patients. BMC Cancer. 2016;16(1):1-8.
17.    Kim JM, Ko H, Kim HJ, Shim SH, Ha Ch, Chang H, et al. Chemopreventive properties of genipin on AGS cell line via induction of JNK/Nrf2/ARE signaling pathway. J Biochem Mol Toxicol. 2016;30(1):45-54.
18.    Sufian HB. Studying the anticancer properties of Parthenolide (PTL) in MCF-7 breast cancer cells . Sufian-Thesis. 2018;8(1):40-88.
19.    Aghaz F, Vaisi Raygani A, Khazaei M, Arkan E, Sajadimajd S, Mozafarie H, et al. Co-encapsulation of tertinoin and resveratrol by solid lipid nanocarrier (SLN) improves mice in vitro matured oocyte/morula-compact stage embryo development. Theriogenology. 2021;171(5):1-13.
20.    Motiei M, S Kashanian. Novel amphiphilic chitosan nanocarriers for sustained oral delivery of hydrophobic drugs. Eur J Pharm Sci. 2017; 9(5):285-291.
21.    Gaonkar RH, Ganguly S, Dewanjee S, Sinha S, Gupta A, Ganguly S, et al. Garcinol loaded vitamin E TPGS emulsified PLGA nanoparticles: preparation, physicochemical characterization, in vitro and in vivo studies. Sci Rep. 2017;7(1):1-14.