Evaluation of PI۳K/AKT, EGFR, P۵۳, Wee۱ genes expression in cervical cancer cells induced by chitosan-cisplatin nanoparticles Enhanced with Gemcitabine

AbstractIntroductopn: Cervical cancer remains one of the leading cancers in the world and is considered one of the worst health issues affecting most developing countries. Infection by human papillomavirus induces critical cellular pathways such as PI۳K/AKT, EGFR, P۵۳, and Wee۱. In the present study, the researchers investigate the effects that chitosan-cisplatin nanoparticles combined with gemcitabine will have on cervical cancer cells in the name of altering the expression of some important genes.Materials and Methods: The current study deals with the preparation, cisplatin and gemcitabine loading on chitosan nanoparticles, cultural manipulations of a HeLa cell line and its treatment using different concentration series of nanoparticles, characterization using UV-Vis Spectroscopy, FTIR and SEM, followed by cytotoxic effects analyzed in the MTT assay and estimation of gene expressions of PI۳K, AKT, EGFR, P۵۳ and Wee۱ by real time polymerase chain reaction.Results: The prepared chitosan nanoparticles had particle sizes that ranged between ۱۸.۶۱ nm to ۲۱۱.۴ nm; besides, there was effective drug loading. The MTT assay displayed that CSNps-CP-Gem caused significant inhibition of viability, with the IC۵۰ values calculated as ۶.۰۰۵ µg/mL and ۴.۰۵۰ µg/mL at ۲۴ and ۴۸ hours, respectively. Gene expression analysis showed a significant down-regulation of PI۳K, AKT, and EGFR to ۰.۷۳۷, ۰.۶۶۴, and ۰.۵۷۸, respectively, while the P۵۳ was upregulated two-fold to ۲.۲۷۴. Wee۱ expression was down-regulated but not at a statistically significant level.Discussion: These results have shown that chitosan-cisplatin nanoparticles, combined with gemcitabine, may effectively enhance the cytotoxicity to cervical cancer cells with very minimal toxicity to normal tissues. Modulation of some key signaling pathways indicates a possible mechanism for the observed therapeutic effects. The present study hence underlines the potential of nanoparticle-based drug delivery systems for the improvement of treatment outcomes in cervical cancer and thus warrants further investigation into their clinical applications.Keywords: PI۳K/AKT, EGFR, P۵۳, Wee۱ genes, cervical cancer,chitosan-cisplatin nanoparticles, gemcitabineAbstractIntroductopn: Cervical cancer remains one of the leading cancers in the world and is considered one of the worst health issues affecting most developing countries. Infection by human papillomavirus induces critical cellular pathways such as PI۳K/AKT, EGFR, P۵۳, and Wee۱. In the present study, the researchers investigate the effects that chitosan-cisplatin nanoparticles combined with gemcitabine will have on cervical cancer cells in the name of altering the expression of some important genes. Materials and Methods: The current study deals with the preparation, cisplatin and gemcitabine loading on chitosan nanoparticles, cultural manipulations of a HeLa cell line and its treatment using different concentration series of nanoparticles, characterization using UV-Vis Spectroscopy, FTIR and SEM, followed by cytotoxic effects analyzed in the MTT assay and estimation of gene expressions of PI۳K, AKT, EGFR, P۵۳ and Wee۱ by real time polymerase chain reaction. Results: The prepared chitosan nanoparticles had particle sizes that ranged between ۱۸.۶۱ nm to ۲۱۱.۴ nm; besides, there was effective drug loading. The MTT assay displayed that CSNps-CP-Gem caused significant inhibition of viability, with the IC۵۰ values calculated as ۶.۰۰۵ µg/mL and ۴.۰۵۰ µg/mL at ۲۴ and ۴۸ hours, respectively. Gene expression analysis showed a significant down-regulation of PI۳K, AKT, and EGFR to ۰.۷۳۷, ۰.۶۶۴, and ۰.۵۷۸, respectively, while the P۵۳ was upregulated two-fold to ۲.۲۷۴. Wee۱ expression was down-regulated but not at a statistically significant level. Discussion: These results have shown that chitosan-cisplatin nanoparticles, combined with gemcitabine, may effectively enhance the cytotoxicity to cervical cancer cells with very minimal toxicity to normal tissues. Modulation of some key signaling pathways indicates a possible mechanism for the observed therapeutic effects. The present study hence underlines the potential of nanoparticle-based drug delivery systems for the improvement of treatment outcomes in cervical cancer and thus warrants further investigation into their clinical applications. Keywords: PI۳K/AKT, EGFR, P۵۳, Wee۱ genes, cervical cancer,chitosan-cisplatin nanoparticles, gemcitabine