Investigating the effect of silica-chitosan nanocomposite encapsulated Epigallocatechin gallate with aptamer decorated on ERK2 and hTERT gene expression in skov3 ovarian cancer cell line

Abstract

introduction Ovarian cancer is the eighth most widespread cause of cancer death and the seventh most widespread cancer in women with five-year survival rates below %45. Epigallocatechin-3-gallate (EGCG) is one of the major polyphenols which exists in green tea and widely used as a chemopreventive agent and anticancer drug which push tumors to the apoptosis. It can prevent the growth of different ovarian cancer cell lines, arrest the cell cycle in G1 or G1/S phase, and regulate the gene expression. nanotechnology scientists have formulated therapeutic drugs in different types of nanoformulations. These can provide targeted drug delivery, improve bio viability, stability, and solubility of therapeutic agents. The main perpose of this study is to investigating the antiproliferative effect of nanocapsulated and pure EGCG and nanocapsulated EGCG which decorated with AS1411 aptamer on the expression level of hTERT and ERK2 genes in the SKOV3 ovarian cancer cell line.method and materials:chitosan coated silica (SiO2@CS) NPs was synthesized and EGCG loaded. SKOV-3 cell line are cultured in a cell culture medium. The antiproliferative and anti-tumor effects of pure and nano-encapsulated EGCG are investigated by MTT assay. The aptemer AS1411 was conjugated to the nanocapsulated EGCG. Total RNA is extracted from the control and treated groups and cDNA is synthesized using the Reverse transcriptase enzyme. Then real-time PCR is performed. Real-time PCR data are compared with control and treated groups and the expression differences in four studied groups are identified. Results: The MTT test results showed that the cytotoxic effects of nano-encapsulated EGCG was more than pure form. RT-PCR Results showed that the groups which treated with nanocapsulated EGCG-Apt has reduced the expression level of hTERT and ERK2 in the SKOV-3 cell line in comparison to the pure and nanocapsulated EGCG. Conclusion: The MTT test results showed 55 that the cytotoxic effects of nano-encapsulated EGCG was more than pure form but the group. SiO2@CS-EGCG-Apt NPs (%51) showed higher internalization efficiency in SKOV-3 cell line than SiO2@CS-EGCG NPs (%29). SiO2@CSEGCG– Apt NPs down-regulated the expression of ERK2 and hTERT on SKOV-3 cells in comparison to the groups which treated with free EGCG and SiO2@CS-EGCG. These results suggested that SiO2@CS-EGCG-Apt had great potential for targeted delivery of different therapeutic agents like EGCG to the SKOV-3 cell line to increase the efficacy of drug delivery and intensify the cytotoxic effect of EGCG.