Evaluation of apoptosis induction in HCT 116 cell line treated with curcumin encapsulated in nano-niosomes
Evaluation of apoptosis induction in HCT 116 cell line treated with curcumin encapsulated in nano-niosomes
Navid Mousazadeh,1Mahmoud Gharbavi,2Hamid Rashidzadeh,3Hamed Nosrati,4Hossein Danafar,5Behrooz Johari,6,*
1. Department of Medical Biotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran 2. Department of Pharmaceutical Biomaterials, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran 3. Cancer Gene Therapy Research Center, Zanjan University of Medical Sciences, Zanjan, Iran 4. Department of Pharmaceutical Biomaterials, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran. 5. Cancer Gene Therapy Research Center, Zanjan University of Medical Sciences, Zanjan, Iran 6. Department of Medical Biotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran
Introduction: Among various types of cancer, colorectal cancer (CRC) ranks third in terms of incidence but second in mortality (1). Chemotherapy, surgery, and radiotherapy are common methods of treatment of CRC (2). Emergence of drug resistance and severe drug toxicity associated with chemotherapeutics have dramatically restricted the successful treatment of cancer disease, thus in this context wise steps should be taken to tackle these problems. Dietary phytochemicals are promising options in cancer therapy owing to their potential in cancer prevention and growth inhibition. Curcumin (CUR) is a dietary phytochemical found in Curcuma longa and has presented a broad range of biological activities, such as anti-oxidant, anti-malarial, anti-inflammatory and anti-cancer effects and exhibited great anti-cancer activity against a variety of different malignancies (3-5).Furthermore, CUR has displayed good pharmacological properties, but due to low absorption, rapid metabolism, low bioavailability and degradability in alkaline conditions have limited its therapeutic application (5). In this regard, researchers have devoted substantial efforts to improving its therapeutic efficacy. One promising approach to overcome the aforementioned drawbacks is using nanotechnology based drug carriers.. Niosomes (non-ionic surfactant vesicles) as an effective novel drug vehicles have been applied to deliver different types of bioactive compounds such as peptides, antigens, hormones, genes and chemotherapeutics (6).The most remarkable feature of niosomes is capability of loading both lipophilic and hydrophilic drugs in which lipophilic one entrapped in membrane bilayer of niosome whereas hydrophilic drugs entrapped into the aqueous core (7). Of note, the benefits of niosomes have not been ended just with the aforementioned advantages, other striking strengths are biodegradability, low toxicity, biocompatibility and non-immunogenicity (8). Within the framework of the above mentioned criteria we aimed at preparing niosome loaded CUR and investigate its potential in induction of apoptosis against human colorectal cancer cell lines HCT116 and compared its efficacy to free CUR
Methods: HCT116 cells were maintained in DMEM High Glucose with 10% fetal bovine serum, penicillin and streptomycin. Typically, HCT116 cells were cultured and incubated in a %5 CO2 atmosphere at 37°C. In brief, HCT 116 cells were seeded in a 12‐well plate, and once the confluency of HCT116 cells reached to the desired amounts, they were treated with the different groups and after incubating for 24 hours; cellular apoptosis was assessed by flow cytometry. It should be noted that the CUR loaded niosomes were prepared by thin-film hydration method and their size was determined by dynamic light scattering (DLS) technique. Standard curve of CUR was obtained by measurement the absorption peak of free CUR at wavelength of 428 nm at different concentrations. All attained data were expressed as the mean ± SD (standard deviation).
Results: DLS measurement indicated that the hydrodynamic diameter of the fabricated niosomal formulation was below 100 nm. In this study, cells were treated with free CUR, blank niosomes and CUR loaded niosomes and after 24 hours of incubation the cell apoptosis were investigated using flow cytometry technique. It was found that both free CUR and CUR loaded niosomes at the same concentration (40 μM) could significantly induce apoptosis in HCT116 cell line. The percentage of apoptosis for free CUR, blank niosomes and CUR loaded niosomes were 16.83 ± 1.50, 5.50 ± 0.87 and 23.80 ± 2.29, respectively
Conclusion: In this study, HCT 116 colon cancer cells were treated with both free CUR and CUR loaded niosomes, and cell apoptosis results showed that the CUR loaded niosomes exhibited significantly higher cell apoptosis than free CUR, which indicating the importance role of using nanocarriers in induction of apoptosis. On the other hand, blank niosomes did not induce significant apoptosis compared to control group and outlining the fact that the fabricated niosomes as a drug vehicles are suitable and practically non-toxic. Overall, nanocarrier-based drug delivery systems can play an important role in inducing cellular apoptosis, but more comprehensive studies should be conducted in this regard to fully elucidate these findings.