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•Adjuvant therapy has been emerged to overcome drug toxicity through diminishing of the drugs side effects.•Stattic is one of the first small molecules recognized to limit STAT3 ...activity through dimerization process.•The implement of stattic triggered anti-proliferative and apoptotic behavior of doxorubicin on ZR-75-1 breast cancer cells.•The interactive effects of stattic and doxorubicin in ZR-75-1 breast cancer cell line have strong synergism.
Adjuvant therapy with novel and effective component has been presented as a contrivance in breast cancer treatment versus the conventional methods. The current research was done to evaluate the implement of stattic, specific STAT3 inhibitor on the anti-proliferative and apoptotic behavior of doxorubicin on ZR-75-1 breast cancer cells. Cell viability was investigated by MTT assay, the percentage of apoptosis by DAPI staining, and Annexin V. Real Time-PCR was applied to find out the correlation between mechanistic roles of the STAT3 pathway and apoptotic signal in the modulation of Bcl-2 and Bax gene expressions axis. The IC50 values for doxorubicin and stattic were 2.5 ± 0.18 μM and 3.5 ± 0.28 μM, respectively. Combination index (CI) value for ZR-75-1 breast cancer was 0.72, which indicated a strong synergistic effect. Incubation of the cells with a combination of stattic and doxorubicin revealed a significant increase in growth inhibitory effect of doxorubicin with more than 50% decrease in proliferation rate and a two-fold increase in the percentage of apoptotic cells. Assessment of gene expression levels demonstrated a visible decrease in antiapoptotic Bcl-2 and Bcl-xl accompanied by an increase in pro-apoptotic Bax mRNA levels (p < 0.05). Taken together, our results show that combination of a STAT3 inhibitor and doxorubicin can be figured out as a promising approach for dealing of patients with breast cancers.
Nowadays, nanoparticle-based combination therapy has been emerging as huge innovation in cancer treatment. Here, we studied the effect of Stattic (STAT3 inhibitor) loaded in nanostructured lipid ...carriers (NLCs) on enhancing the efficacy, cytotoxicity, and induction of apoptosis of doxorubicin in B16F10 mouse melanoma cancer cell. The evaluation of Stattic-loaded NLCs has been done in terms of zeta potential, particle size, scanning electron microscope (SEM), and cellular uptake. MTT assay was applied to evaluate the cell proliferation. Apoptotic cell death and identification of early and late apoptosis were assessed by DAPI staining and Annexin V/PI staining, respectively. Real-time RT-PCR was applied to measure the effects of doxorubicin and/or Stattic on key apoptotic genes such as Bad, Survivin, HIF1, and STAT3. The Stattic formulated into NLCs shown mean particle size of 56 ± 7 nm which was confirmed by SEM. The IC
50
values for Stattic and doxorubicin were 2.95 ± 0.52 μM and 1.21 ± 0.36 μM, respectively. Stattic-loaded NLCs diminished percent of cell proliferation from 68 ± 6.8 to 54 ± 3.7% (
p
< 0.05). Combinational treatment of the cells with Stattic-loaded nanoparticles and doxorubicin give rise to a significant increase in the percentage of apoptosis (
p
< 0.05). The study of gene expression profile has shown a remarkable decrease in anti-apoptotic gene, Survivin, along with smooth decline in HIF1 as angiogenesis intermediator and increase in Bad mRNA levels. Our results recommend that NLCs as novel technology have potent strategy to augment efficacy of current chemotherapeutic agent in melanoma cancer cells.
The aim of this study was to evaluate the potential of zoledronic acid (ZOL)-loaded lipidic nanoparticles (ZOL-NLCs) in enhancing the efficiency of paclitaxel (Pac) in the context of cytotoxicity, ...apoptosis, and invasiveness of HepG
2
hepatocellular carcinoma cells. ZOL-NLCs were characterized in terms of zeta potential, particle size, and scanning electron microscope (SEM) as well as cell internalization. To measure the anti-proliferative effects of ZOL-NLCs, annexin-V/PI and MTT assays were employed. Real-time PCR and western blot analysis were performed to identify the molecular mechanisms underlying the apoptosis in response to the studied conditions. Furthermore, the transwell migration assay was applied to clarify the role of applied formulations on the invasiveness of HepG
2
cells. Our results demonstrated that the optimized ZOL had an average particle size of 105 ± 6 nm with a nearly narrow size distribution. The IC
50
values for ZOL and ZOL-NLCs were 90 ± 3.1 and 54.6 ± 2.4 µM, respectively. The population of apoptotic cells was increased from 17 ± 2% to 27 ± 4% (
p
< 0.05) in response to treatment with ZOL-NLCs. ZOL-loaded nanoparticles triggered the mRNA expression of Bax as pro-apoptotic marker and E-cadherin as epithelial one along with a decrease in mesenchymal marker, N-cadherin, and Bcl-xl as an anti-apoptotic marker in HepG
2
cells. These outcomes were consistent with western blot analysis of protein expressions. Besides, ZOL-incorporated lipidic nanoparticles reduced the migration of HepG
2
cells significantly. Our data suggest that the formulation of ZOL into lipidic nanoparticles can be considered a potential therapeutic approach that can enhance the efficacy of Pac chemotherapy.
Nuclear factor erythroid 2-related factor 2 (Nrf2) is believed to be responsible for the control mechanisms of cellular defense response and master regulator of antioxidant system by adjustment of ...endogenous antioxidants, phase II detoxifying enzymes and transporters, so inhibition of Nrf2 could be considered molecule target to overcome drug resistance and cancer progression. By harnessing liposome as an advanced nanoparticles transporter, we formulated Quinacrine known as nrf2 inhibitor into nano-carrier, and sensitized A-549 lung tumor cells to Cisplatin. The aim of this work was to prepare liposome nano-carriers to enhance the bioavailability of Quinacrine and to improve passive targeting in A549 cells. Quinacrine formulation into liposome exposed a mean particle size of 80±5 nm in passive targeting and 110±3 after decoration with chitosan oligosaccharides (COS), respectively. The highest amount of cell death (
p<0
.05) occurred with the co-incubation of the A549 cells with new formulation and Cisplatin. Additionally, Quinacrine-loaded liposomes declined Nrf2 expression more than Quinacrine alone (
p
<0.05). Correspondingly, the expression of Nrf2 downstream genes, MRP1, Trx, and bcl2 decreased significantly. Taking all the data into consideration, liposomes containing Quinacrine could ameliorate the effectiveness of Cisplatin by raising the permeability of cancer cells to the abovementioned chemical treatment and might be then given as a candidate to boost the therapeutic protocols in cancer patients.
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•Melatonin increases efficacy of tamoxifen in induction of apoptosis in MCF-7 cells.•Nano structured Lipid Carriers prepared with particle size of 71nm.•Melatonin-loaded NLCs had ...synergistic effects on cytotoxic effects of tamoxifen.•NLCs can be considered as an advanced delivery system to increase cellular uptake and bioavalibility of anti-cancer agents.
Finding advanced anti-cancer agents with selective toxicity in tumor tissues is the goal of anticancer delivery systems. This study investigated potential application of nanostructured lipid carriers (NLCs) in increasing melatonin induced cytotoxicity and apoptosis in MCF-7 breast cancer cells.
Melatonin-loaded NLCs were characterized for particle size, zeta potential, Fourier transforms infrared spectroscopy, differential scanning calorimetry, cellular uptake, and scanning electron microscope (SEM). Anti-proliferative and apoptotic effects of new formulation were evaluated by MTT and flow cytometric assays, respectively. Gene expression of apoptotic markers including survivin, Bcl-2 and Bid were examined by Real time quantitative PCR.
The optimized formulation of NLCs revealed mean particle size of 71±5nm with nearly narrow size distribution. The formulation exhibited an acceptable stability during four months in terms of size and lack of drug release. The IC50 values for melatonin and tamoxifen were 1.3±0.4mM and 30.7±5.2μM, respectively. Melatonin loaded NLCs decreased percentage of cell proliferation from 55±7.2% to 40±4.1% (p<0.05). Co-treatment of the cells with melatonin loaded nanoparticles and tamoxifen caused two fold increase in the percentage of apoptosis (p<0.05). Evaluation of gene expression profile demonstrated a marked decrease in anti-apoptotic survivin with increase in pro-apoptotic Bid mRNA levels.
Taken together, our results suggest NLC technology as a promising delivery system, which elevates the efficacy of chemotherapeutics in breast cancer cells.
The objective of this study was construction of recombinant hEGF-pPIC9 which may be used for expression of recombinant hEGF in following studies.
EGF cDNA was purchased from Genecopoeia Company and ...used for PCR amplification. Prior to ligation, the PCR product and pPIC9 vector was digested with EcoRI and XhoI and ligated in pPIC9 vector and subjected to colony PCR screening and sequencing analysis.
PCR amplification of EGF cDNA using recombinant hEGF-pPIC9 vector as template was concluded in amplification of 197bp fragment. Construction of recombinant hEGF-pPIC9 of EGf gene was verified by PCR and sequencing.
Construction of Recombinant hEGF-pPIC9 was the primary stage for production and expression of EFG in the future study.
In this study, a transmission line model is applied to the electrochemical impedance spectroscopy (EIS) data of the fabricated dye-sensitized solar cells (DSSCs) to evaluate the charge transfer ...mechanism through the cells. Natural dye from black plum (Syzygium cumini) fruit was used as a cell sensitizer (SC-DSSC) and compared its photovoltaic and electron transport capabilities to those of a cell using a synthetic sensitizer (N719-DSSC). TiO2/ZnO electrospun composite nanofibers were used as the semiconductor layer of the photoanode to enhance electron transfer. The EIS analysis revealed the role of electron resistances through shant, interfaces, and electrolyte solution by measuring the electron transfer kinetic parameters of each element. Based on the results, the SC-DSSC and N719-DSSC are appropriate photovoltaic cells because their ratios of effective electron diffusion length to photoanode thickness are 12.5 and 2.8, respectively. The EIS analysis showed that the electrospun composite nanofiber coated on the photoanode reduces the semiconductor layer's electrical resistance to the cell's total resistance. The extracted natural dye also boosted electron lifetime to 3.68 ms and diffusion coefficient to 54.3×10−6 m2/s while minimizing back-electron recombination at the semiconductor-electrolyte interface. Moreover, the semiconductor-electrolyte interface resistance is over 85% of the overall resistance for both DSSCs and controls electron transport through the cells, which is due to the dye-semiconductor binding intensity. Based on the photovoltaic data, the SC-DSSC cell efficiency was lower than N719-DSSC which is attributed to its higher electron transfer rate-controlling element. Thus, enhancing dye-semiconductor interactions will decrease the rate-controlling impedance and enhance cell performance.
Bovine serum albumin (BSA) decorated lipidic core nanoparticles were constructed for the delivery of quinacrine (Quin) to enhance docetaxel (Dtx) dose-dependent behavior against A-549 lung cancer ...cells through the inhibition of the Nrf2 signal pathway and arresting the Sub-G1 cell cycle mechanism. Chitosan was chosen as a suitable stabilizer in addressing the proton sponge effect with BSA providing the favorable protein corona formation along with low toxicity. The optimized formulation was characterized in terms of particle size, zeta potential, morphology, polydispersity index and encapsulation efficiency (EE). The mechanism of cytotoxicity, apoptosis and cell cycle arrest along with the determination of the combined efficacy of Quin with Dtx were investigated by MTT assay, flow cytometric assay, CompuSyn and HSA synergy score analysis. Real-time PCR and Western blot analysis were performed to quantify Nrf2-dependent genes and protein levels, respectively. The results demonstrated an average particle size of 112 ± 16.2 nm, a positive surface charge of 15 ± 0.1 mV along with an EE of 65.4 ± 2.6%. Quin NPs reduced the proliferation of lung cancer cells from 68 ± 6.1% to 49 ± 4.5% (p < 0.05) along with a two-fold surge in the percentage of apoptosis (p < 0.05). The incubation of A549 lung cancer cells with Quin NPs caused a significant decrease in Nrf2, NQO1 and MRP1 associated with an increase in BAD mRNA levels compared to the other groups (p < 0.05). Combinational treatment of both Quin + Dtx, as well as Quin NPs + Dtx, induced a remarkable G2M arrest and sub-G1 apoptosis. The results, therefore, indicate that Quin-loaded nanoparticles have the ability to enhance the anti-proliferation efficacy of Dtx against A549 lung cancer by inhibiting Nrf2, increasing synergistic interaction with lower doses of Dtx and arresting cell cycle in the Sub-G1 thus leading to the induction of apoptosis.
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•Melanoma is a possibly lethal skin malignancy.•WYE-132 is specific inhibitor of mTORC1 and mTORC2 signaling pathways.•in vitro studies showed significant down-regulation in ...anti-apoptotic genes.•WYE-132 and vinblastine presented synergistic anticancer effects.
This study aimed to evaluate the synergistic anticancer effect of vinblastine and WYE-132, a specific inhibitor of the mammalian target of rapamycin (mTOR), on B16F10 melanoma cancer cells. MTT assay, Annexin V, and DAPI staining along with Real-Time PCR were conducted to understand the mechanistic roles of mTOR pathway in melanoma cancer cells. The IC50 values for vinblastine and WYE-132 were 39.4 ± 1.8 nM and 145.2 ± 4.5 nM, respectively. The co-administration of WYE-132 and vinblastine in B16F10 cells offered a significant increase in the growth inhibitory effect of vinblastine along with a two-fold escalation in the percentage of apoptotic cells. The calculation of gene expression levels confirmed a visible fall in anti-apoptotic Bcl-2, Ki-67 and Mcl-1 accompanied by a surge in pro-apoptotic Bax mRNA levels (p < 0.05). Thus, we showed that the combination of WYE-132 and vinblastine could be a promising approach for dealing with patients with melanoma cancers.
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•ATRA increases efficacy of doxorubicin in induction of apoptosis in MDA-MB-231 cancer cells.•Nano structured Lipid Carriers obtained with particle size of 95nm.•ATRA-loaded NLCs had ...synergistic effects on cytotoxicity effects of doxorubicin.•NLCs pave the ways for advanced drug delivery to enhance apoptosis and bioavailability of chemotherapy agents.
Drug delivery-based nanoparticles have been emerged to be an alternative and efficient approach to cancer therapy compared to conventional systems. Here, we investigated the role of all-trans retinoic acid (ATRA) formulated with precirol in increasing doxorubicin (Dox) induced apoptosis and cell cycle arrest in MDA-MB-231 breast cancer cells.
ATRA-loaded Nano structured lipid carriers (NLCs) were evaluated in terms of particle size, zeta potential, Fourier transforms infrared spectroscopy (FTIR), cell internalization, and scanning electron microscope (SEM). To understand molecular mechanism of apoptosis and cell cycle progression flow cytometric assay, MTT and DAPI staining was applied. Real time (RT)-PCR analysis was employed to investigate the expression of apoptosis related genes, including Survivin, Bcl-2 and Bax.
The optimized ATRA formulation exhibited average particle size of 95±5nm with nearly narrow size distribution. The IC50 values for ATRA and doxorubicin were 48±0.4μM and 0.81±0.02μM, respectively. ATRA-loaded NLCs decreased percentage of cell proliferation from 51±7.2% to 36±4.1% (p <0.05). Co-treatment of the MDA-MB-231 cells with ATRA formulation and doxorubicin caused two-fold increase in the percentage of apoptosis (p<0.05). The results from gene expression exhibited a significant decrease in survivin along with increase at Bax mRNA levels accompanied by a slight increase in Bax/Bcl-2 ratio.
Our results propose that ATRA encapsulated in precirol as a biocompatible compound augments the efficacy of Dox in cancer therapy.
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