Cytotoxic Effect of Doxorubicin Loaded on Nanocarrier Systems on Human Cervical Cancer (HeLa) Cell Line and Determination of Some Genes Expression Profiles

Abstract Doxorubicin (DOX) is one of the drugs that used in treatment of cervical cancer. DOX has a wide spectrum of anticancer activity, but its clinical application is limited because of its serious side effects, especially cardiotoxicity. In this study, firstly titanium dioxide (TiO₂) nanoparticles were synthesized in order to reduce or completely remove such disadvantages and increase the anticancer activity of DOX. Due to the very high toxicity of TiO₂, a new drug delivery system called nanocarrier system, was created when it TiO₂ modified with polyethylene glycol (PEG). Finally, the synthesized nano-carrier system was modified with anti-cancer drug DOX. In this study, the synthesized nanoparticles TiO₂, DOX, PEG-TiO₂, PEG-TiO₂-DOX were applied on human cervical cancer cell line (HeLa) and cytotoxic dose was determined by MTT method. Different concentrations (0.5–50 µg / ml) of TiO₂, DOX, PEG-TiO₂, PEG-TiO₂-DOX drugs were applied on the human cervical cancer cell line HeLa for 24, 48 and 72 hours, and IC₅₀ doses were calculated. Among these drugs, TiO₂ and DOX were found to be the most active after 72 hours. PEG-TiO₂-DOX nanocarrier based drug was found to be IC₅₀ 6.467 ± 0.135 µg / ml. In the next step, obtained from the incubation of each drugs for 48 hours IC₅₀ doses were applied on HeLa cell line and incubated for 48 hours then RNA was izolated from the cells. After that we synthesized cDNA from RNA samples and 24 genes were determined by real time RT-PCR analysis method. As a result, all gene expression increased with the effect of TiO₂ drug except GPX1, PRDX1, CTNNB1 and CDNK1A genes. With the effect of PEG-TiO₂, the expression levels of GPX1, SOD1, CAT, PRDX1, CSNK1A1, TOP2A, BAX, NQO1, TP53 and CASP2 genes increased, while the expression levels of all other genes had decreased. On the other hand, while the expression level of CTNNB1 and MYC genes had decreased with the effect of DOX, the expression levels of other genes had increased. When the effect of PEG-TiO₂-DOX was examined, we found that the expression levels of all genes had increased except ERCC1, ATR, PRKDC, PDGFB and MYC genes expression had decreased.