The effect of nanoparticle quercetin loaded on the h-tert and pinx1 gene expressions and induction of opoptosis in breast canser cells (MCF7)

Abstract

Cancer is one of the most important health issues in the modern world. Breast cancer ranks first among women. Common treatments, Therefore, new approaches to cancer treatment such as nanotechnology and the use of natural compounds have received much attention. Therefore, the aim of the present study was to investigate the effect of pure quercetin drug loaded on MPEG-PCL nanoparticles on the expression of hTERT and pinx1 genes and induction of apoptosis on breast cancer cell line (MCF7). Materials and Methods: mPEG-PCL nanoparticles were fabricated using a suitable percentage of both materials. Quercetin was loaded into nanoparticles at different concentrations. The amount of drug loaded was measured. Quercetin release from nanoparticles was measured at pH 5.5 and pH 7.5. The effect of the prepared nanosystem on MCF-7 cancer cells was investigated. The cells were cultured in complete culture medium and treated in different groups of quercetin, mPEG-PCL nanoparticles and mPEG-PCL nanoparticles containing quercetin. MTT method was used to evaluate cell viability. Flow cytometry was used to evaluate cell apoptosis and finally the expression of Bcl2, Bax, hTERT and Pinx1 genes was evaluated by RT-PCR. Results: The mean diameters of mPEG-PCL and mPEG-PCL nanoparticles loaded with quercetin were 50.65±1.8 and 66.56 ±1.4 nm, respectively, as well as the Z potential of mPEG-PCL and mPEG-PCL nanoparticles loaded quercetin was -14.8 and -14.3 mV, respectively. The drug loading rate was 6.7%. In the first 24 hours, the release rate of the drug at acidic pH was about 13%, while at normal pH it was about 5%. MTT results showed that quercetin and quercetin-containing nanoparticles had the greatest effect on cell viability after 48 hours. Based on the results, the rate of apoptosis was 2.85% in the control group, 8.29% in the quercetin group, 7.5% in the nanoparticles group and 18.14% in the drug-containing nanoparticles group. Based on the results, Bcl2 gene expression in the mPEG-PCL loaded quercetin had a significant decrease compared to the group of nanoparticles, quercetin (p = 0.0001) and control (p = 0.05). Also, expression of Bax gene in cells treated with mPEG-PCL loaded quercetin had a significant increase compared to control (p = 0.0001). The expression of hTERT gene in the mPEG-PCL loaded quercetin was significantly reduced compared to the control group (p = 0.05) and the nanoparticle group (p = 0.0001). hTERT was also significantly reduced in the quercetin group compared to the control group (p = 0.0001). Pinx1 gene was significantly increased in the mPEG-PCL loaded quercetin compared to other groups (p = 0.0001) and in the quercetin group was significantly increased compared to the control group (p = 0.05).