Preparation and characterization of nanoparticles containing farnesol and evaluation of their in vitro anti-cancer effects in colorectal cancer
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Atefeh Ebrahimi,1 Neda Mousavi-Niri,2,* Maryam Naseroleslami,3 Leila YouseftTabar,4
1. Department of Biotechnology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
2. Department of Biotechnology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
3. Department of Cellular and Molecular Biology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
4. Department of Organic Chemistry , Faculty of Pharmaceutical Chemistry, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
- Introduction: The progression of colon cancer is mainly characterized by a rapid increase in tumor mass and volume (in solid tumors), an invasive increase, and an overall decrease in patient survival. In cancer, autophagy plays a dual role in preventing tumor growth by inhibiting the accumulation of damaged proteins and, on the other hand, promotes tumor growth by helping its cells survive. In tumor therapy, apoptosis tolerance is an important mechanism of tumor resistance to treatment. Autophagy can prevent drug-induced apoptosis and tumor resistance. However, autophagic cell death may also be a form of death for apoptotic tolerant tumor cells. Farnesol is an isoprenoid alcohol obtained from various plants such as citronella,cyclamen, roses, and musk. Its anti-neoplastic effects in various types of human cancers, such as breast cancer, lung cancer, prostate cancer, pancreatic cancer and multiple myeloma, have been investigated by inhibiting cell proliferation in vitro and suppressing tumor growth in vivo. On the other hand, niosomes, which are one of the new drug delivery systems, have received a lot of attention today due to their better penetration and controlled release in position, biodegradable, and biocompatibility with the body. There is currently no effective treatment for tumors among the conventional therapies used because most conventional therapies kill cancer cells but also damage healthy tissue. This study aimed to evaluate the effect of farnesol-bearing nanoniosome in the treatment of colon cancer.
- Methods: Niosome nanoparticles containing farnesol were prepared using a thin-layer hydration method containing a mixture of Span60, and cholesterol in different molar ratios. All formulations were identified in terms of size, entrapment efficiency, release specifications. Sampling was performed for soluble farnesol and niosome containing farnesol for up to 72 hours during specified times (1, 2, 4, 8, 24, 48, and 72). The effect of farnesol-containing nanocarriers on the HT29 cell line of colon cancer was investigated by MTT assay and cell cycle by flow cytometry. The effect of drug containing nanocarriers on the expression of LC3 and ATG genes involved in autophagy was also investigated by Real-Time PCR
- Results: According to the morphology of nanoparticles with a SEM microscope, the particles have a uniform spherical structure and suitable size. The optimal sample with a lipid to drug ratio of 30 and a surfactant to cholesterol ratio of 2 was selected under 7 min sonication time. The drug retention efficiency in the sample optimized by the box-Behnken method is 94.57 ± 1.24. The average size of nanoparticles without Farnesol is 209 nm with a dispersion index of 0.7, which increases to 233 nm when the drug is introduced into the nanoparticle. In the first 6 hours, approximately 85% of soluble farnesol is released and after 48 hours, the release reaches 100%. While the release of the Farnesol-bearing Niosome is a two-step process, the first step is the rapid phase, with 50% of the drug released in the first 24 hours, but after 72 hours the drug release reaches almost 60%. Farnesol-containing nanoniosomes significantly (Pvalue˂0.001) reduced the cytotoxicity of the compound and showed higher efficacy than the free drug. The results of this study showed that niosomal nanocarriers containing farnesol had a lethal effect on cancer cells in 72 hours at all concentrations.
Flowcytometric studies of Annexin V-PI showed that cell death, early apoptosis of HT29 cells treated with IC50 nanoparticles containing farnesol increased by about 12% after 48 hours compared to the control group. The results of cell cycle analysis showed that about 24% of the cells were treated in the sub G1 stage as a result of treatment with biological nanoparticles. The increase in cell accumulation in the sub G1 stage compared to the control group was statistically significant (Pvalue˂0.001). On the other hand, a significant decrease was seen in the cell population located in the G2 stage compared to the control group. In the expression of genes involved in autophagy (LC3, ATG) increase in expression was seen compared to the control group (Pvalue˂0.001).
- Conclusion: According to the results of this study, Niosome containing farnesol with desirable characteristics induced apoptosis; Autophagy in cancer cells and increase the lethal effect on cancer cells. As a conclusion, it can be a good candidate for the treatment of colon cancer.
- Keywords: Farnesol ,colorectal cancer, Autophagy
