Evaluating the Effect of target miR-۱۰۱ transcripts (ZEB۱) Plasmid on Lung Cancer A۵۴۹ Cell Apoptosis

Background and Aim: Despite significant advancements in the area of oncology, lung cancer still remains one of the most malignant cancers in terms of incidence and mortality. Therefore, special attention must be paid to finding new alternative treatments. This study aimed to evaluate the effect of miR-۱۰۱ plasmid on lung cancer A۵۴۹ cells’ apoptosis and migration. To this end, after testing apoptosis and evaluating cellular migration the expression rate of target miR-۱۰۱ transcripts (ZEB۱) assessed by real-time PCR. According to the results of the study, the ZEB۱ gene expression decreased after the transfer of miR-۱۰۱ to lung cancer cells. Therefore, it can play a critical role and apoptosis.Methods: Nano Particles synthesis was carried out based on the previously published protocol and miR-۱۰۱-containing plasmid preparation Plasmid extraction steps were performed according to the Geneall plasmid extraction.The nano-drop apparatus was exploited to determine the extracted plasmid concentration. The cells in each well were treated using specific concentrations of the desired method. The culture medium of cells was changed ۴۸ hours after treatment, and the MTT combination was added to cells, followed by incubation in dark for four hours.The absorbance of the samples was carried out at ۵۷۰ nm with a reference filter of ۶۳۰ nm using ELISA Plate Reader. In addition, the results were analyzed based on Real-Time PCR and Apoptosis assessment using Annexin/PI kit by flow cytometry.Results: Analysis of real-time PCR results: the test was carried out to evaluate the expression of miR-۱۰۱ genes (ZEB۱)in lung cancer cell lines. According to the test results, the increase of miR-۱۰۱ gene expression decreased the expression of ZEB۱, thereby inducing apoptosis and cellular death. The results showed that ۱۱.۷۴% apoptosis and ۷.۶۷% necrosis were observed in the treated cells. Given the relatively low lethal impact of miRs, this level of apoptosis lethality in the study was acceptable, and it could be concluded that the method could treat patients in the future.Conclusion: In this study, the NPs were entered into the cells and the target gene was released, which led to target gene silencing.Therefore, apoptosis induction and decreased number of migrated cancer cell lines were observed.

کلیدواژه ها:

Gene Therapy ، miR-۱۰۱ ، Nanocarrier ، Lung Cancer