Co-delivery of doxorubicin and conferone by novel pH-responsive β-cyclodextrin grafted micelles triggers apoptosis of metastatic human breast cancer cells

Abstract

Adjuvant-aided combination chemotherapy is one of the most effective ways of cancer treatment by overcoming the multidrug resistance (MDR) and reducing the side-effects of anticancer drugs. In this study, Conferone (Conf) was used as an adjuvant in combination with Doxorubicin (Dox) for inducing apoptosis to MDA-MB-231 cells. Herein, the novel biodegradable amphiphilic β-cyclodextrin grafted poly maleate-co-PLGA was synthesized by thiol-ene addition and ring-opening process. Micelles obtained from the novel copolymer showed exceptional properties such as small size of around 34.5 nm, CMC of 0.1 μg/mL, and cell internalization of around 100% at 30 min. These novel engineered micelles were used for combination delivery of doxorubicin-conferone with high encapsulation efficiency of near 100% for both drugs. Our results show that combination delivery of Dox and Conf to MDA-MB-231 cells had synergistic effects (CI < 1). According to cell cycle and Annexin-V apoptosis analysis, Dox-Conf loaded micelle significantly induce tumor cell apoptosis (more than 98% of cells population showed apoptosis at IC50 = 0.259 μg/mL). RT-PCR and western-blot tests show that Dox-Conf loaded βCD-g-PMA-co-PLGA micelle induced apoptosis via intrinsic pathway. Therefore, the unique design of multi-functional pH-sensitive micelles open a new perspective for the development of nanomedicine for combination chemo-adjuvant therapy against malignant cancer.

Enhanced anticancer potency with reduced nephrotoxicity of newly synthesized platin-based complexes compared with cisplatin

“…Evaluation of cell cycle progression was performed by flow cytometry of propidium iodide and can identify the fraction of time spent by cells in various stages of the cell cycle. First, A549, MCF-7, and Caco-2 cells were seeded in 6-well plates at 25*10 4 cells per well and exposed to a concentration at the IC 50 of Pt(TSC)Cl complex and cisplatin for 48 h. Cell cycle distribution was determined by flow cytometry by the protocol described previously 28 . The control condition consisted of cells cultured in the absence of treatment but in the presence of PI.…”

“…For evaluation of the apoptosis pathway genes, real time PCR was carried out based on the following PCR program 28 : initial denaturation at 95 °C for 15 min, 45 cycles of denaturation at 95 °C for 15 sec, and annealing/extension at 60 °C for 50 sec. The real time PCR mixture contained 5 μL 2x SYBR Green Master Mix (RealQ Plus 2x Master Mix Green, Ampliqon, Denmark), 2 μL cDNA, 0.5 μL of 5 pmol/μL primer pair mix (Eurofin, Germany), and 3 μL H2O.…”

“…Caco-2 cancer cells, as the most sensitive cells to Pt(TSC)Cl, were seeded in 6-well plates at a population of 10 6 cells per well and treated with cisplatin and Pt(TSC)Cl for 48 h. Assessment of apoptotic protein level alterations by western blot analysis by the protocol described previously 28 . The control included cells without treatment.…”

Cyclodextrin-Based Polymeric Drug Delivery Systems for Cancer Therapy

“…More than 98% of cells showed apoptosis after Dox and Conf coloaded micelle treatment at a dose of IC 50 (0.259 μg/mL). The intrinsic mitochondrial apoptosis pathway (p53, p27, Bcl-2/Bax, cleave-caspase-9, cleaved caspase-7 and cleaved caspase 3) may account for the Dox and Conf coloaded micelle-induced apoptosis [ 162 ].…”

Development of Solid Lipid Nanoparticles with Letrozole for Breast Cancer Treatment: in-vitro and in-vivo evaluation

“…In addition, BC cells may develop resistance to treatment [4].As effective alternatives to chemotherapy drugs, nano-cancer drugs are an excellent alternative to conventional anti-tumor therapies. [5].…”