Human Cervical Carcinoma HeLa Cells Research Articles

An automated approach for hemocytometer cell counting based on image-processing method

Evaluating cell viability plays an important role in routine cell culture, toxicology, and drug development. Trypan blue stain is one of the simplest assays that are used to determine the number of viable cells in a cell suspension. The stained cell suspension can then be counted under a microscope using a hemocytometer. Hemocytometers have a fixed volume, making it possible to calculate the concentration of cells in the sample and measure viability. Most of the related studies are conducted using manual counting methods. However, manual counting is time-consuming and laborious. While various automated cell counting devices have been proposed, they tend to be too expensive for widespread deployment. Although low-cost automated cell counting methods presented in the literatures provide high accuracy and cost-effective alternatives, only cells in the field of view of microscope with unknown volume are counted. This will make the concentration inestimable. Accordingly, the present study proposes a low-cost but accurate method for hemocytometer cell counting using an image-processing approach. The accuracy of the proposed method is demonstrated by comparing the cell counting results obtained for human cervical carcinoma (HeLa) cells with the manual counts. It is shown that the proposed approach has a mean absolute percentage error of less than 6.26 % for samples containing a mixture of live and dead cells. Moreover, the proposed system is equipped with a simple and intuitive human machine interface such that the counting task can be performed by individuals with no prior experience in the image-processing field.

The Anti-proliferative Effect of Caffeic Acid and Dactolisib on Human Cervical Carcinoma HeLa Cell Line

Cervical carcinoma is a common gynecological cancer with high mortality rate among women worldwide. Caffeic acid exerts an antiproliferative effect against cervical carcinoma. Dactolisib is a dual PI3K and mTOR inhibitor that has a therapeutic potential for cervical carcinoma. This study aimed to reveal the anti-proliferative effect of combination treatment of caffeic acid and Dactolisib on cervical carcinoma HeLa cell line. Cytotoxicity of caffeic acid and Dactolisib on HeLa cell line was assessed by MTS assay. Colony formation of HeLa cells treated with caffeic acid and Dactolisib was determined by staining colonies with crystal violet and visualizing under light microscope. Dactolisib decreased cell proliferation of HeLa cells in time and dose dependent manner. 5 μM caffeic acid did not show any significant change in cell viability of HeLa cells. Combination treatment of 5 μM caffeic acid and 0.5 μM Dactolisib showed decrease in cell viability of HeLa cells when compared to Dactolisib treated cells. Combination of caffeic acid and Dactolisib decreased colony diameter of HeLa cells significantly when compared to control group. Caffeic acid and Dactolisib shows anti-proliferative effect on human cervical carcinoma HeLa cell line, so further studies should be performed to reveal the mechanism of action.

A Study on the Biological Activity of Optically Pure Aziridine Phosphines and Phosphine Oxides.

A series of optically pure aziridine phosphines and their corresponding phosphine oxides were synthesized through established chemical methodologies. The compounds were systematically investigated for their biological properties. Notably, all synthesized compounds demonstrated moderate antibacterial activity only against the reference strain of Staphylococcus aureus. However, compounds 5 and 7 exhibited noteworthy cell viability inhibition of human cervical epithelioid carcinoma HeLa cells and endometrial adenocarcinoma Ishikawa cells. Further studies of these compounds revealed additional biological effects, including disruption of the cell membrane in high concentrations, cell cycle arrest in the S phase, and the induction of reactive oxygen species (ROS). Comparative analysis of the two classes of chiral organophosphorus derivatives of aziridines indicated that chiral phosphine oxides displayed significantly higher biological activity. Consequently, these findings suggest that chiral phosphine oxides may be potential candidates for the development of anticancer drugs. In light of the significant interest in preparations whose structure is based on a three-membered aziridine ring in terms of potential anticancer therapy, this research fits into the current research trend and should constitute a valuable addition to the current state of knowledge and the existing library of aziridine derivatives with anticancer properties.

Exploring In-Silico, In-Vitro Antioxidant, and Cytotoxic Potential of Valerian wallichii by On Cervical Epithelial Carcinoma (HeLa) Cell Lines.

Traditional medicinal practices often utilize herbal remedies for treating various diseases. This study focuses on exploring the phytochemical constituents, in-silico, in-vitro antioxidant, and anticancer activities of Valerian wallichii root extracts on human cervical epithelial carcinoma (HeLa) cell lines. The molecular docking approach was employed to predict the ligand molecule's orientation within the receptor like Epidermal growth factor receptor tyrosine kinase domain (PDB ID: 1M17) using Schrodinger's GLIDE model. Among the selected phytocompounds, hesperidin exhibited promising inhibitory activity against EGFR (Epidermal Growth Factor Receptor) domain with -8.701 kcal/mol docking score and interactions with MET 769, ASP 831, ASP776, LEU694 and ASN818 residues as compared to standard doxorubicin with -7.6 kcal/mol docking score and interactions with ASP 831, ASN818 and ASP776 residues and further, various antioxidant activity was assessed and In-vitro anticancer activity against HeLa cell lines was evaluated by hydroalcoholic root extracts demonstrated antioxidant capacities, and selective cytotoxicity was observed, with IC50 : 45.759±0.42 μg/mL for the overall extract and IC50 : 30.245±0.58 μg/mL for the EAF fraction as compared to standard doxorubicin with IC50 : 25.9891±0.25 μg/mL, respectively. The present study concluded that Valerian wallichii L possesses potential human cervical epithelial carcinoma cell line inhibition properties based on the computer aided drug design models and in-vitro activity.

Cell cycle dependence of cell survival following exposure to X-rays in synchronous HeLa cells expressing fluorescent ubiquitination-based cell cycle indicators.

The cell cycle dependence of radiosensitivity has yet to be fully determined, as it is technically difficult to achieve a high degree of cell cycle synchronization in cultured cell systems and accurately detect the cell cycle phase of individual cells simultaneously. We used human cervical carcinoma HeLa cells expressing fluorescent ubiquitination-based cell cycle indicators (FUCCI), and employed the mitotic harvesting method that is one of the cell cycle synchronization methods. The imaging analysis confirmed that the cell cycle is highly synchronized after mitotic cell harvesting until 18-20 h of the doubling time has elapsed. Also, flow cytometry analysis revealed that the S and G2 phases peak at approximately 12 and 14-16 h, respectively, after mitotic harvesting. In addition, the clonogenic assay showed the changes in surviving fractions following exposure to X-rays according to the progress through the cell cycle. These results indicate that HeLa-FUCCI cells become radioresistant in the G1 phase, become radiosensitive in the early S phase, rapidly become radioresistant in the late S phase, and become radiosensitive again in the G2 phase. Our findings may contribute to the further development of combinations of radiation and cell cycle-specific anticancer agents.

The Fatty Acid Composition and the MTT-Cytotoxicity Test of Commercially Available Commiphora Gileadensis Balsam Oil

Commiphora gileadensis, an aromatic plant, is traditionally used in Middle Eastern countries to prevent pain and inflammation. Fatty acid composition, peroxide number and free fatty acidity analyzes of commercially available C. gileadensis balsam oil and investigation of its effect on cell viability. Commercially available C. gileadensis balsam oil fatty acid composition, peroxide number and free fatty acidity values were determined by the IUPAC IID19 method, and MTT cell viability tests were performed on L929 fibroblast and HeLa (human epithelial cervical carcinoma) cell lines. Commercially available C. gileadensis balsam oil did not exert cytotoxic effects on both L929 fibroblast and HeLa (human epithelial cervical carcinoma) cell lines and promoted the growth of cell lines. Due to its growth promoting feature on L929 fibroblast cells, which is a healthy cell line, this material can be used as a cell culture medium additive.

Identification by methods of steady-state and kinetic spectrofluorimetry of endogenous porphyrins and flavins sensitizing the formation of reactive oxygen species in cancer cells.

The question about acceptor molecules of optical radiation that determine the effects of photobiomodulation in relation to various types of cells still remains the focus of attention of researchers. This issue is most relevant for cancer cells, since, depending on the parameters of optical radiation, light can either stimulate their growth or inhibit them and lead to death. This study shows that endogenous porphyrins, which have sensitizing properties, may play an important role in the implementation of the effects of photobiomodulation, along with flavins. For the first time, using steady-state and kinetic spectrofluorimetry, free-base porphyrins and their zinc complexes were discovered and identified in living human cervical epithelial carcinoma (HeLa) cells, as well as in their extracts. It has been shown that reliable detection of porphyrin fluorescence in cells is hampered by the intense fluorescence of flavins due to their high concentration (micromolar range) and higher (compared to tetrapyrroles) fluorescence quantum yield. Optimization of the spectral range of excitation and the use of extractants that provide multiple quenching of the flavin component while increasing the emission efficiency of tetrapyrroles makes it possible to weaken the contribution of the flavin component to the recorded fluorescence spectra.

Glyoxylate reductase: Definitive identification in human liver mitochondria, its importance for the compartment-specific detoxification of glyoxylate.

Glyoxylate is a key metabolite generated from various precursor substrates in different subcellular compartments including mitochondria, peroxisomes, and the cytosol. The fact that glyoxylate is a good substrate for the ubiquitously expressed enzyme lactate dehydrogenase (LDH) requires the presence of efficient glyoxylate detoxification systems to avoid the formation of oxalate. Furthermore, this detoxification needs to be compartment-specific since LDH is actively present in multiple subcellular compartments including peroxisomes, mitochondria, and the cytosol. Whereas the identity of these protection systems has been established for both peroxisomes and the cytosol as concluded from the deficiency of alanine glyoxylate aminotransferase (AGT) in primary hyperoxaluria type 1 (PH1) and glyoxylate reductase (GR) in PH2, the glyoxylate protection system in mitochondria has remained less well defined. In this manuscript, we show that the enzyme glyoxylate reductase has a bimodal distribution in human embryonic kidney (HEK293), hepatocellular carcinoma (HepG2), and cervical carcinoma (HeLa) cells and more importantly, in human liver, and is actively present in both the mitochondrial and cytosolic compartments. We conclude that the metabolism of glyoxylate in humans requires the complicated interaction between different subcellular compartments within the cell and discuss the implications for the different primary hyperoxalurias.

Cytotoxic and Antioxidant Properties of Artemisia deserti Essential Oil Obtained by Different Extraction Methods

Background: Artemisia deserti Krasch belongs to Asteraceae family, and has many medicinal properties that used to treat a variety of diseases, including antihypertensive, invigorating blood circulation, antiallergy, antiviral, antitumor, and antioxidant. Methods: The objective of this study was the evaluation of cytotoxic and antioxidant activities as well as the chemical composition of A. deserti essential oil extracted by different extraction methods. These extraction methods included hydro-distillation (HD), salt-HD (Salt pretreatment followed by HD), maceration-HD (maceration pretreatment followed by HD), acid-HD (acid pretreatment followed by HD), ultrasound-HD (ultrasound pretreatment followed by HD), and enzyme-HD (enzyme pretreatment followed by HD). Results: The results revealed that the highest yield of essential oil was achieved by acid-HD (0.48% ± 0.11%). In all methods, the main compounds of essential oil were camphor (44.32%–66.80%), piperiton (14.11%–24.33%), and 1,8-cineole (4.85%–6.75%). The antioxidant property of essential oils was investigated using the DPPH method. Based on the results, the essential oils extracted by acid-HD and enzyme-HD methods had the highest property. The cytotoxic property of A. deserti essential oils was evaluated. The results demonstrated that the essential oil extracted by enzyme-HD indicated the highest cytotoxicity activity against human cervical carcinoma (HeLa) cells in both 24 and 48 h. Based on the gas chromatography-mass spectrometry results, 1,8-cineol, carvacrol, thymol, and myristicin were rich in the essential oil extracted by enzyme-HD, and the highest cytotoxicity activity of essential oil obtained by enzyme-HD method is probably related to these compounds. Conclusion: In addition, it was observed that enzyme-HD is an effective method in the extraction of essential oil with the highest antioxidant and cytotoxic activities.

Diastereoselective 1,3-dipolar cycloaddition of cyclopropenes to acenaphthenequinone azomethine ylides

A diastereoselective 1,3-dipolar cycloaddition of azomethine ylides generated from acenaphthylene-1,2-dione (or aceantrylene-1,2-dione) and amino acids with 1,2-diphenylcyclopropenes has been developed. As a result, cyclopropa[a]pyrrolizidines and 3-azabicyclo[3.1.0]hexanes spiro-fused to acenaphthylen-1(2H)-one and aceanthrylen-1(2H)-one fragments were obtained for the first time. Based on DFT calculations at M11/cc-pVDZ level of theory, a concerted mechanism of cycloaddition was proposed to explain the observed diastereoselectivity. The cytotoxic activity of newly synthesized compounds against human erythroleukemia (K562) and human cervical carcinoma (HeLa) cell lines was evaluated in vitro by MTS-assay.

Validation of mitotic harvesting method with human cervical carcinoma HeLa cells expressing fluorescent ubiquitination-based cell cycle indicators for radiation research.

The cell cycle is a series of events in the process of one cell giving rise to two daughter cells. The mitotic harvesting method, established by Terasima and Tolmach in the 1960s, causes minimal physiological stress on the cells and achieves a high degree of cell cycle synchrony by collecting only mitotic cells from a cultured cell system. The purpose of the present study is to validate the versatility of the mitotic harvesting method using human cervical cell line HeLa cells expressing Fluorescent Ubiquitination-based Cell Cycle Indicators (FUCCI) and to estimate the cell cycle-dependent changes in radiosensitivity in HeLa-FUCCI cells. The image analysis showed that cell cycle synchrony was maintained for at least 24 hours after mitotic cell collection. Also, the clonogenic assay demonstrated changes in radiosensitivity that were cell cycle dependent. These results indicate that the mitotic harvesting method using FUCCI-expressing cells has high versatility in the field of radiation cell biology.

Diminazene aceturate-induced cytotoxicity is associated with the deregulation of cell cycle signaling and downregulation of oncogenes Furin, c-MYC, and FOXM1 in human cervical carcinoma Hela cells.

Diminazene aceturate (DIZE)is an FDA-listed small molecule known for the treatment of African sleeping sickness. In vivo studies showed that DIZE may be beneficial for a range of human ailments. However, there is very limited information on the effects of DIZE on human cancer cells. The current study aimed to investigate the cytotoxic responses of DIZE, using the human carcinoma Hela cell line. WST-1 cell proliferation assay showed that DIZE inhibited the viability of Hela cells in a dose-dependent manner and the observed response was associated with the downregulation of Ki67 and PCNA cell proliferation markers. DIZE-treated cells stained with acridine orange-ethidium and JC-10 dye revealed cell death and loss of mitochondrial membrane potential (Ψm), compared withDMSO (vehicle) control, respectively. Cellular immunofluorescence staining of DIZE-treated cells showed upregulation of caspase 3 activities. DIZE-treated cells showed downregulation of mRNA for G1/S genes CCNA2 and CDC25A, S-phase genes MCM3 and PLK4, and G2/S phase transition/mitosis genes Aurka and PLK1. These effects were associated with decreased mRNA expression of Furin, c-Myc, and FOXM1 oncogenes. These results suggested that DIZE may be considered for its effects on other cancer types. To the best of our knowledge, this is the first study to evaluate the effect of DIZE on human cervical cancer cells.

Development of new cobalt, copper, and zinc complexes of Schiff-base ligands as prospective chemotherapeutic agents

Fluorobenzaldehyde was converted into new, bidentate Schiff-base ligands with nitrogen and sulphur donor sites in two steps. The first step proceeds via Knoevenagel condensation; the product obtained was treated with thiosemicarbazide to yield the ligand L1. The Schiff-base ligand, L2, was synthesized by treating L1 with 4-chlorobenzylchloride in a basic medium. Co (II), Cu (II), and Zn (II) complexes were synthesized with the ligands L1 and L2. All these complexes were characterized by FTIR, UV–Vis, NMR spectroscopy, ESI-mass spectral analysis, magnetic moment, molar conductance, thermal, powder XRD, and electrochemical studies. From these studies, the molecular formulae of the complexes were found to be [M(L)2Cl2], where M = Co (II), Cu (II) and Zn (II) and L = L1 or L2 based on the ligand used in the synthesis. The synthesized complexes were observed to have a high spin and distorted octahedral geometry and were non-electrolytic. The UV absorption titrimetric analysis confirmed that these complexes would bind strongly to CT-DNA through the intercalative mode. The electrophorograms supported the statement that Cu (II) complexes could effectively catalyze the hydrolytic cleavage of CT-DNA and can be considered potent metallonuclease agents. The DPPH assay determined the antioxidant properties of these complexes, and the cupric complexes were observed to possess better antioxidant properties than the standard used ascorbic acid. The cell viability analysis of these complexes showed an elevated control of proliferation against a human breast adenocarcinoma (MCF-7) cancer cell line compared to a human cervical carcinoma (HeLa) cell line. The results exhibited by these compounds were compared with the standard drug, cisplatin. The binding affinity of the Cu (II) metal ion complex towards CT-DNA was high when compared to Co (II) and Zn (II) metal ions. These results are in line with the cytotoxicity studies. Molecular docking studies also hold up the experimental data. Cu (II) complexes using ligand L2 show higher efficiency than those with ligand L1 in all biological assays and can be considered a potent DNA-targeting drug.

Alternaria alternata F3, a Novel Taxol-Producing Endophytic Fungus Isolated from the Fruits of Taxus cuspidata: Isolation, Characterization, Taxol Yield Improvement, and Antitumor Activity.

In this study, a novel taxol-producing endophytic fungus, strain F3, was isolated from the fruits of Taxus cuspidata and identified as Alternaria alternata according to its macroscopic and microscopic traits and sequence analysis of internal transcribed spacer (ITS). The presence of taxol was detected by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) and confirmed by ultra-high-performance liquid chromatography-electrospray coupled to tandem mass spectrometry (UPLC-ESI-MS/MS) and nuclear magnetic resonance (NMR). The fermentation parameters of strain F3 were then optimized for high taxol production. The maximum taxol yield of 195.4µg L-1 by A. alternata F3 was observed in 200-mL yeast peptone dextrose (YPD) broth, at an initial pH value of 6.0, supplemented with 0.1g L-1 sodium acetate, 0.25g L-1 salicylic acid, and 0.00125g L-1 silver nitrate and inoculum size 2%, and incubated at 28°C and 150rpm for 8days, which was 2.12-fold compared with the initial yield of taxol. Also, fungal taxol exhibited antitumor activity towards human lung carcinoma (A549) cell line and human cervical carcinoma (Hela) cell line with IC50 values of 3.98µgmL-1 and 0.35µgmL-1. Overall, this is the first report on taxol-producing endophytic fungus isolated from the fruits of Taxus. This study offers a novel source for the production of taxol for anticancer treatment.

Different Behavior of 2-Substituted 3-Nitro-2H-chromenes in the Reaction with Stabilized Azomethine Ylides Generated from α-Iminoesters.

The AgOAc-catalysed reaction of 3-nitro-2-phenyl-2H-chromenes with stabilized azomethine ylides generated from the imines based on methyl glycinate and arylaldehydes leads to a mixture of endo and endo' isomers of the corresponding chromeno[3,4-c]pyrrolidines in a ratio of 2.0-2.3:1 in 85-93% total yields as a result of a Michael addition/Mannich reaction sequence. In a similar reaction involving 2-trifluoromethyl-3-nitro-2H-chromenes, only endo chromeno[3,4-c]pyrrolidines are formed in 85-94% yields. 3-Nitro-2-(trichloromethyl)-2H-chromenes under the same conditions react with these azomethine ylides to give the corresponding Michael adducts as individual anti-isomers with the cis,trans-configuration of the chromane ring in 40-67% yields. Some 4-CF3-substituted chromano[3,4-c]pyrrolidines exhibited high cytotoxic activity against HeLa human cervical carcinoma cells.

Catechin, rutin and quercetin in Quercus mongolica Fisch leaves exert inhibitory effects on multiple cancer cells.

We aimed to identify anti-tumor agents in Quercus mongolica Fisch (QMF). Bioactive compounds in QMF leaves, which were extracted using ethanol as a co-solvent. Five point zero six grams of flavonoids were obtained from 100 g of QMF leaves. Catechin (18.4%), rutin (6.3%), ellagic acid (34.9%), quercetin (5.1%) and kaempferol (20.6%) are the main ingredients of the extracts and were further purified by HPLC. CCK-8 cell proliferation assay showed that catechin and ellagic acid exerted strong inhibitory effects on the proliferation of all cancer cells with lower IC50 values against MCF-7 human breast cancer cell lines, SMMC-7721 human hepatocellular carcinoma cells, HeLa human cervical carcinoma cell lines and SKOV3 human ovarian carcinoma cell lines (p < .05). Catechin, rutin and quercetin induced a higher rate of apoptosis and inhibited all cancer cell proliferation by inducing the G0/G1 phase and G2/M phase arrest (p < .05). However, ellagic acid induced tumor cell death, not through apoptosis and there may be other molecular mechanisms. High levels of catechin and ellagic acid in QMF can be developed as potential drugs to treat different types of cancer cells. PRACTICAL APPLICATIONS: Quercus species have been widely studied because of their antioxidant, anti-inflammatory, antimicrobial, and anti-tumor properties. Bioactive compounds in the leaves of Quercus mongolica Fisch have high levels of catechin and ellagic acid, which exert significant inhibitory properties on the proliferation of various types of cancer cells. Therefore, the bioactive compounds may be potential natural drugs in the prevention of cancer development and progression.

Synthesis, Cellular Uptake, and Photodynamic Activity of Oligogalactosyl Zinc(II) Phthalocyanines.

A series of di-α-substituted zinc(II) phthalocyanines with different number of galactose moieties, ranging from 1 to 8, namely Pc-galn (n=1, 2, 4, and 8) were designed and synthesized. The synthesis involved the copper-catalyzed azide-alkyne cycloaddition reaction of a mono- or dialkynyl zinc(II) phthalocyanine with an acetyl-protected galactosyl azide or its dendritic derivative with four acetyl-protected galactosyl groups, followed by removal of the acetyl protecting groups via alkaline hydrolysis. In N,N-dimethylformamide, these oligogalactosyl phthalocyanines were non-aggregated as shown by the strong Q-band absorption and fluorescence emission. Owing to the di-α-substitution, they also behaved as efficient singlet oxygen generators upon light irradiation with a singlet oxygen quantum yield of 0.84. The spectroscopic and photophysical properties were not affected by the number of galactosyl units. In contrast, the compounds became significantly aggregated and quenched in phosphate-buffered saline. Their cellular uptake was then studied using a range of cell lines, which generally followed the order Pc-gal1 >Pc-gal2 ≈Pc-gal4 >Pc-gal8 . Interestingly, the di-galactosyl analogue exhibited selective uptake against HeLa human cervical carcinoma cells through an energy-dependent pathway instead of the expected asialoglycoprotein receptor. Upon light irradiation, it could effectively kill the cells with a half-maximal inhibitory concentration of 0.58 μM.

Prodigiosin R3, a new multicyclic prodigiosin formed by prodigiosin cyclization genes in the roseophilin producer.

The rph gene cluster for prodigiosin biosynthesis has been identified in Streptomyces griseoviridis 2464-S5, which produces cyclic prodigiosin derivatives including roseophilin (2), prodigiosin R1 (3) and prodigiosin R2 (4). A new cyclic prodigiosin, prodigiosin R3 (1), was produced by the redG redP double disruptant of Streptomyces coelicolor M511 expressing four cyclization gene candidates (rphG, rphG2, rphG3 and rphG4) in the rph cluster. The same compound was isolated from Streptomyces griseoviridis 2464-S5. The molecular formula of 1 was established as C27H33N3O by ESI and FAB mass spectrometry. The structure was determined to be a multicyclic prodigiosin with three alkyl linkages by NMR spectroscopic analysis. Prodigiosin R3 (1) showed cytotoxicity against HeLa human cervical carcinoma cells and HT1080 human fibrosarcoma cells with IC50s of 2.1 µM and 3.2 µM, respectively.

Redox-sensitive hyaluronic acid-ferrocene micelles delivering doxorubicin for enhanced tumor treatment by synergistic chemo/chemodynamic therepay

In order to solve the defects of chemotherapeutics, numerous collaborative therapy systems have been developed according to the characteristics of tumor microenvironment. Herein, a targeting redox-sensitive micellar system (DOX/FCH) composed of ferrocene (Fc) and hyaluronic acid (HA) was prepared to deliver doxorubicin (DOX) for synergetic chemotherapy and chemodynamic therapy (CDT). DOX/FCH could accurately target the tumor site through high affinity between HA and high-expressed CD44 receptors in human cervical carcinoma (HeLa) cells. The appropriate particle size made it easier for cells to take up the micelles. The DOX released from DOX/FCH reached about 50% in 2 h, because the disulfide bonds were depolymerized in reducing environment which simulate the tumor intracellular environment. Moreover, the production of hydroxyl radical (·OH) was detected both extracellular and intracellular, which indicated that FCH could exert the CDT effect through Fenton reaction to enhance the chemotherapy of DOX. The good biocompatibility in 3T3 cells as well as the obvious cytotoxicity in HeLa cells demonstrated that FCH had the possibility of becoming an excellent nanocarrier. Also, the cytotoxicity of DOX/FCH also confirmed the synergetic effect of CDT of FCH and chemotherapy of DOX. Therefore, DOX/FCH shows great potential in the application of improving the efficacy of combined chemotherapy and CDT.

Combination Treatment of Retinoic Acid Plus Focal Adhesion Kinase Inhibitor Prevents Tumor Growth and Breast Cancer Cell Metastasis.

All-trans retinoic acid (RA), the primary metabolite of vitamin A, controls the development and homeostasis of organisms and tissues. RA and its natural and synthetic derivatives, both known as retinoids, are promising agents in treating and chemopreventing different neoplasias, including breast cancer (BC). Focal adhesion kinase (FAK) is a crucial regulator of cell migration, and its overexpression is associated with tumor metastatic behavior. Thus, pharmaceutical FAK inhibitors (FAKi) have been developed to counter its action. In this work, we hypothesize that the RA plus FAKi (RA + FAKi) approach could improve the inhibition of tumor progression. By in silico analysis and its subsequent validation by qPCR, we confirmed RARA, SRC, and PTK2 (encoding RARα, Src, and FAK, respectively) overexpression in all breast cells tested. We also showed a different pattern of genes up/down-regulated between RA-resistant and RA-sensitive BC cells. In addition, we demonstrated that both RA-resistant BC cells (MDA-MB-231 and MDA-MB-468) display the same behavior after RA treatment, modulating the expression of genes involved in Src-FAK signaling. Furthermore, we demonstrated that although RA and FAKi administered separately decrease viability, adhesion, and migration in mammary adenocarcinoma LM3 cells, their combination exerts a higher effect. Additionally, we show that both drugs individually, as well as in combination, induce the expression of apoptosis markers such as active-caspase-3 and cleaved-PARP1. We also provided evidence that RA effects are extrapolated to other cancer cells, including T-47D BC and the human cervical carcinoma HeLa cells. In an orthotopic assay of LM3 tumor growth, whereas RA and FAKi administered separately reduced tumor growth, the combined treatment induced a more potent inhibition increasing mice survival. Moreover, in an experimental metastatic assay, RA significantly reduced metastatic lung dissemination of LM3 cells. Overall, these results indicate that RA resistance could reflect deregulation of most RA-target genes, including genes encoding components of the Src-FAK pathway. Our study demonstrates that RA plays an essential role in disrupting BC tumor growth and metastatic dissemination in vitro and in vivo by controlling FAK expression and localization. RA plus FAKi exacerbate these effects, thus suggesting that the sensitivity to RA therapies could be increased with FAKi coadministration in BC tumors.