Synthesis and Biological Activity of Diastereomeric and Geometric Analogs of Calcipotriol, PRI-2202 and PRI-2205, Against Human HL-60 Leukemia and MCF-7 Breast Cancer Cells

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HLA-G is a nonclassical MHC Class I molecule that protects the fetus from the maternal immune system, and it also plays a role in immune evasion of cancer. Previously we have shown that iron can suppress the cytolytic function of natural killer cells to human breast cancer cell lines. We have found that breast cancer cells over express HLA-G mRNA and proteins in both cell lines and breast carcinomas. In order to determine if iron concentration affects HLA-G expression, we cultured the human breast cancer cell line MCF-7 in media with 100 mM, 400 mM, and 1600 mM of iron for 24 hours. We used RT-PCR to examine HLA-G mRNA expression of the MCF-7 cells. The PCR primers were designed so that they could detect all alternatively spliced HLA-G mRNA isoforms, 1000 bp (HLA-G1 and HLA-G5), 600 bp (HLA-G2 and HLA-G4), and 300 bp (HLA-G3). RT-PCR showed that the MCF-7 cells cultured with control medium only had one band of 300 bp mRNA expression. However, the MCF-7 cells cultured in medium containing 100 mM, 400 mM, and 1600 mM of iron had three bands of 300 bp, 600 bp, and 1000 bp HLA-G mRNA expression. Moreover, the intensity of the 300 bp band from the iron-treated MCF-7 cells is significantly higher than the cells cultured with control medium. In summary, iron upregulates the HLA-G mRNA expression of MCF-7 cells, and this probably contributes to cancer immunosuppression and to the cytolytic dysfunction of natural killer cells. Citation Format: Jonathan F. Head, Xian P. Jiang, Robert L. Elliott. Iron upregulates HLA-G mRNA expression of the human MCF-7 breast cancer cell line. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1507. doi:10.1158/1538-7445.AM2013-1507

ST14 (suppression of tumorigenicity 14) is a transmembrane serine protease that contains a serine protease catalytic (SP) domain, an SEA domain, two complement subcomponent C1r/s (CUB) domains, and four low density lipoprotein receptor class A domains. Glutathione S-transferase fusion proteins with SP, CUB, and low density lipoprotein receptor domains and their corresponding mutants were generated to analyze protein interactions with these domains. Modified glutathione S-transferase pull-down assays demonstrated the interaction between the SP domain and hepatocyte growth factor activator inhibitor-1. With the same method, a CUB domain-interacting protein was isolated and turned out to be the transmembrane protein with epidermal growth factor-like and two follistatin-like domains 1 (TMEFF1). Quantitative real time PCR revealed that the expression of the TMEFF1 gene was dependent on the transfection of the ST14 gene in the RKO cell line. Our results also suggested that ST14 and TMEFF1 were co-expressed in the human breast cancer cell line MCF7, human placenta, kidney, and liver tissues. Interestingly, these two genes were co-up-regulated in kidney tumors versus normal tissues, consistent with our results that showed the dependence of TMEFF1 expression on ST14 in RKO cells. Finally, homology modeling studies suggested that TMEFF1 might form a complex with ST14 by an interaction between epidermal growth factor and CUB domains.

Our previous study has confirmed that IL-7δ5 (an IL-7 variant lacking exon 5) promotes breast cancer growth. However, whether IL-7δ5 is involved in tumor cell EMT and metastasis remains unclear. In this study, we investigated the preclinical effects and molecular mechanisms of IL-7δ5 on EMT and metastasis in human MCF-7 and BT-20 breast cancer cells in vitro and in vivo. The results showed that IL-7δ5 induced EMT and invasion in tumor cells, associated with up-regulation of N-cadherin and the down-regulation of E-cadherin. Furthermore, we found that IL-7δ5 induced the activation of Akt. Inhibition of PI3K/Akt pathway by LY294002 reversed the EMT transition in breast cancer cell lines MCF-7 and BT-20 induced by IL-7δ5. In addition, IL-7δ5 enhanced cancer metastasis and shortened survival time, with increased level changes of activated Akt in nude mice with breast cancer. In conclusion, our findings demonstrate that IL-7δ5 induces human breast cancer cell lines EMT and metastasis via activation of PI3K/Akt pathway. Thus, IL-7δ5 may be a potential target against human breast cancer.

Triorganotin compounds induce hormonal alterations, i.e., endocrine-disrupting effects in mammals, including humans. Tributyltin chloride (TBT-Cl) and triphenyltin chloride (TPT-Cl) are known to function as nuclear retinoid X receptor (RXR) agonists. Their cytotoxic effects in ER(+) luminal human breast cancer cell line MCF-7 and ER(-) basal-like human breast cancer cell line MDA-MB-231 were examined. We observed significantly higher toxicity of TBT-Cl in comparison with TPT-Cl in both cell lines. Comparable apoptosis-inducing concentrations were 200 and 800nM, respectively, as shown by PARP cleavage and FDA staining. Both compounds activated executive caspases in the concentration-dependent manner in MDA-MB-231 cells, but the onset of TPT-Cl-induced caspase-3/7 activation was delayed in comparison with TBT-Cl. Both compounds slowed down the migration of these highly invasive cells, which was accompanied by RARbeta upregulation. Other RAR and RXR expressions were differentially modulated by studied organotins in both cell lines.

Breast and cervical cancers are the most common gender-specific cancers affecting women worldwide. In this investigation, we highlighted the synthesis, VEGFR-2 and p38α MAPK inhibitory activity of new series of fluorinated coumarin-based derivatives featuring a variety of bioactive chemical moieties attached or fused to the coumarin nucleus at the 3 and/or 4 position. The bioactive inhibitors were further assessed for their anti-proliferative effect against human MCF-7 breast cancer and HeLa cervical cancer cell lines, respectively. Most of the tested compounds showed potent preferential inhibition effects against human VEGFR-2 and remarkable anticancer activities in the human breast cancer cell line MCF-7. Compounds 29, 24, and 2 displayed the highest inhibitory activity against VEGFR-2 (94% inhibition) and they were the most potent anticancer agents toward MCF-7 cancer cells with IC50 values of 7.90, 8.28, and 8.30 μg/mL, respectively. Compound 13 inhibited p38α MAPK phosphorylation with a significant reduction in % cell viability against HeLa cancer cells at 10 and 30 µM. Docking experiments carried out on VEGFR-2 and p38 MAPK crystallographic structures revealed that the active compounds bind to the active sites through H-bonds, arene-cation, and hydrophobic π-π interactions. QSAR analysis demonstrated considerable correlation coefficient (R2 = 0.76969) and root mean square error (RMSE = 0.10446) values. Also, the residual values between the experimental pIC50 and predicted pIC50 are very close, indicating the reliability of the established QSAR model.

The aim of the study was to investigate possible effects of p57 on the growth of the human MCF-7 and rat SHZ-88 breast cancer cell lines. Specific oligonucleotide sequences containing small hairpin structure were inserted into a small interfering RNA (siRNA) expression vector. The human MCF-7 and rat SHZ-88 breast cancer cell lines were transfected with recombinant plasmids. The p57 gene expression was blocked in the human MCF-7 breast and rat SHZ-88 breast cancer cells, using chemically modified siRNA. The p57 expression level was evaluated using quantitative polymerase chain reaction (qPCR) and western blot analysis. Immunofluorescence was conducted to detect p57 expression in the breast cancer cells. Tetrazolium blue (MTT) method was employed to detect the effect of p57 inhibition on the proliferation of the MCF-7 and SHZ-88 cell lines. Cell proliferation in the experimental group was significantly reduced. Immunofluorescence assay results showed p57 siRNA effectively inhibited the p57 level in the MCF-7 and SHZ-88 cells. RT-PCR results showed that 48 h after transfection, the p57 mRNA level in the transfected group was significantly lower compared with the control group. In conclusion, p57 effectively inhibited the proliferation of breast cancer after stable interference.

Arene-ruthenium(II) complexes with tetracyclic oxime derivatives: synthesis, structure and antiproliferative activity against human breast cancer cells

Breast cancer is the most frequent malignant tumor among women and is the leading cause of death from cancer worldwide. In 2020, 90,222 deaths from breast cancer were reported in México. However, cancer cells resistant to antineoplastic drugs are frequent, which compromises the patient’s survival. For this reason, in oncology, it is essential to search for new sources of anticancer compounds. Endophytic fungi are currently considered potential reservoirs for compounds with antitumor activity. More than 100 different classes of secondary metabolites have been reported with activity against different types of cancer, including breast cancer. Therefore, the objective of this study was to evaluate the activity of methanol extracts of endophytic fungi isolated from Lophocereus marginatus against the MCF-7 breast cancer cell line, using as controls the MA-104 cell line human peripheral blood mononuclear cells. Fungal strains were isolated from stems of L. marginatus and molecularly identified from ribosomal DNA internal transcript spacer region sequencing. Extraction of the secondary metabolites was performed from the maceration of mycelium fungus in methanol. The solvent was removed with a rotary evaporator and the extract was reconstituted with dimethyl sulfoxide (DMSO). Next, cells were incubated with the extract at concentrations ranging from 31.25 µg/mL to 250 µg/mL for 48 h at 37 °C and 5% CO2. Growth inhibition was assessed by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide reduction assay, using 1% DMSO was used as a negative control. IC50 values and selectivity indexes (SI) were then calculated. It was found that tumor cells growth inhibition by the extracts increased in a concentration-dependent manner. A. versicolor PME-H005 strain extract showed the highest antitumor activity, with up to 58.9% growth inhibition at 250 µg/mL and IC50 value of 95.21 ± 1 µg/mL against MCF-7 cells. The highest SI was obtained with MCF-7 cell s with 2.77, as compared with normal PBMC with an IC50 of 264 ± 1.5 µg/mL. In addition, the M. anisopliae PME-H007 strain presented a high SI value of 2.1 and an IC50 of 245.9 ± 1.9 µg/mL, using the MCF-7 cell line. This study shows the potential of the endophytic fungus A. versicolor PME-H005 isolated from L. marginatus for production of secondary metabolites with antitumor activity against MCF-7 cells. Citation Format: Jesica M. Ramírez Villalobos, Cesar I. Romo Sáenz, Karla S. Morán Santibañez, Patricia Tamez Guerra, Orquídea Pérez González, Reyes Tamez Guerra, Cristina Rodríguez-Padilla, Ricardo A. Gomez Flores. In vitro antitumoral activity of endophytic fungi isolated from Lophocereus marginatus against the human breast cancer cell line MCF-7 [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P3-07-27.

Objective:This work aims to investigate the effect of PXDIOI,a novel potent histone deacetylase inhibitor,on the cell proliferation,cycle arrest and apoptosis of human breast cancer cell line MCF-7 and to preliminarily explore its molecular mechanism. Methods:MCF-7 cells were cultured in RPMI 1640 medium supplemented with 10%fetal bovin serum and were treated with PXDIOI at varying concentrations.The methyl thiazolyl tetrazolium(MTT ) assay and clonogenic assay were used to measure cell proliferation. Morphological changes of cells were observed by fluorescent microscope after staining by Hoechst33342.Flow cytometer was used to analyze the cell cycle arrest rates(PI staining ) and the cell apoptotic rates(AnnexinV-FITC/PI double-staining ).The protein expressions of p21,CyclinB1,PARP,Bcl-2 and Bax were detected by Western blot.Results:PXD101 was used to inhibit the proliferation of the MCF-7 cell line in a dose and time-dependent manner.Fluorescence microscope showed there were nuclear fragmentation and apoptosis bodies in the cells.Flow cytometric analysis indicated that PXD101 induced MCF-7 cells in G_2/M phase were significantly increased.After MCF-7 cells exposed to different concentrations of PXD101,i.e.,0,0.1,1 and 10μmol/L,for 24 h,the ratio of G_2/ M-phase cells was(12.66±1.55 )%,(20.63±1.32 )%,(23.20±1.82 )%and(32.19±2.37 )%respectively(P 0.05 ).The rates of apoptotic cells were also significantly increased,compared with the control group(P 0.05 ).PXDIOI could up-regulate the protein expression of p21 and down-regulate the expression of CyclinBl.The cleavage of PARP and the expression of pro-apoptosis protein Bax were increased while the anti-apoptosis protein Bcl-2 was decreased.Conclusion:PXD101 in vitro can significantly inhibit the proliferation and can induce cell cycle arrest and apoptosis on human breast cancer MCF-7 cell line in a dose-dependent manner.PXD101 may become a new anti-tumor drug for human breast cancer.

The epidermal growth factor receptor (EGFR) (ErbB1) and HER-2/neu (ErbB2) are members of the ErbB family of receptor tyrosine kinases. These receptors are overexpressed in a variety of human tumors and overexpression generally correlates with poor prognosis and decreased survival. Lapatinib, a reversible inhibitor of both EGFR and HER-2/neu, has shown some success in achieving clinical responses in heavily pretreated advanced cancer patients. GW2974 is a reversible dual inhibitor similar to lapatinib, but GW2974 was not progressed to clinical trials due to pharmacokinetic issues. Bcl-2, an anti-apoptotic protein, is also overexpressed in a number of human tumors. Bcl-2 inhibitors induce apoptosis and sensitize cancer cells to other therapies. The purpose of this study was to assess the effects of combining ErbB and Bcl-2 inhibitors on the growth of human breast cancer cell lines. EGFR/HER-2/neu tyrosine kinase inhibitors (lapatinib and GW2974) were combined with Bcl-2 inhibitors (HA14-1 or GX15-070) and the anti-proliferative effects were determined by the MTT tetrazolium dye assay. Combinations were tested in MCF-7 human breast cancer cells, a HER-2/neu transfected MCF-7 cell line (MCF/18), and a tamoxifen-resistant MCF-7 cell line (MTR-3). A synergistic inhibitory effect was observed with the combination of inhibitors of EGFR-HER-2/neu (lapatinib or GW2974) and Bcl-2 (GX15-070 or HA14-1) on the growth of the MCF-7, MCF/18, and MTR-3 human breast cancer cell lines. This study suggests that simultaneously blocking the ErbB family of receptor tyrosine kinases and Bcl-2 family of proteins may be a benefit to breast cancer patients.

According to reports, a serine protease inhibitor (Maspin) suppresses metastasis, invasion and angiogenesis in breast and prostate cancers. Silibinin is a natural polyphenolic flavonoid with anti-cancer activity. We assessed the effects of silibinin on cell viability, maspin and ERα gene expression in MCF-7 cell line. The human MCF-7 breast cancer cell line was cultured in Dulbecco's Modified Eagle's Medium (DMEM) and treated with different concentrations of silibinin (100-600 μg/mL) for 24, 48 and 72 hours. The cytotoxic effect of silibinin on MCF-7 viability was determined using Methyl-Thiazolyl-Tetrazolium (MTT) assay by IC50 determination. The fold changes of Maspin and ERα expression were determined by reverse-transcription real-time Polymerase Chain Reaction (PCR). All experiments on the cells were performed in triplicates. The maximum inhibitory effect of silibinin on cell viability was observed at 600 μg/mL after 72-hour incubation (p = 0.001). Incubation of the cells with silibinin for 48 and 72 hours significantly decreased IC50 values to 250 and 207 μg/mL (p = 0.005 and p= 0.006), respectively. The expression of maspin and ERα in the treated cells compared to controls was significantly decreased following treatment with different concentrations of silibinin during a 24-hour period. Silibinin reduces both maspin and ERα gene expression in MCF-7 cell line. The therapeutic effect of silibinin on the treatment of breast cancer may be mediated by the reduction of ERα expression. For verifying this hypothesis and the possible therapeutic implication of silibinin on breast cancer, further studies in this direction are necessary.

Searching for new active molecules against human breast cancer cell line MCF-7, novel quinoline based thiazolidinones has been efficiently synthesized under ultrasound irradiation. The newly synthesized compounds were tested against human breast cancer cell line MCF-7. Compounds P3, P4, and P6 were found to be promising inhibitors of MCF-7 characterized by lower IC50 values in a dose-dependent mode with high specificity against MCF-7 (IC50 of 10 μM at 24 h). Among all the synthesized compounds, P3, P4, and P6 shows IC50 values 5.38, 5.12, and 0.73 µM, respectively, were considered as a potential lead. These lead molecules showed significant anti-cancer activity against human breast cancer cell line MCF-7. Additionally, induction of G2/M cell arrest within 24 h was discovered via flow cytometry analysis. Overall, our data suggest that potent compounds have an inhibitory effect on cell proliferation of MCF-7 through cell cycle arrest, giving it great potential as a future therapeutic reagent for cancers.

Background: Mammalian target of rapamycin (mTOR) is a kinase pathway that regulates the cell cycle progression and growth. Rapamycin inhibits this pathway. The useful effects of rapamycin on cell growth have been widely shown in animal studies. However, its beneficial effects are associated with some success in benign and malignant cancers, which have produced its moderate outcomes in the clinic. Objectives: The aim of this study was to investigate whether rapamycin can induce oxidative stress in MCF-7 and MDA MB-231 human breast cancer cell lines. Methods: The MCF-7 and MDA MB-231 cell lines were cultivated and treated with rapamycin for 72 hours. The viability of the cells was determined using the colorimetric MTT assay. Lipid peroxidation (TBARS), protein oxidation (carbonyl groups), total antioxidant capacity assay, and glutathione (GSH) levels were measured in the MCF-7 and MDA MB-231 cells both with and without rapamycin treatment. Results: The IC50 concentration of rapamycin was 100 nM in MCF-7 cells, whereas the MDA-MB-231 cells were highly resistant to rapamycin. Our data indicated an increase in oxidative status by increasing lipid peroxidation and protein oxidation, GSH, and total antioxidant capacity levels in the MCF-7 and MDA-MB-231 cell lines exposed to rapamycin in comparison with control cells. Conclusions: These outcomes support our theory that rapamycin increases oxidative stress in MCF-7 and MDA MB-231 cells but also shows high levels of antioxidant effects, which probably limit the effects of the rapamycin on the same issue in the clinic.

To investigate the effects of ribonucleotide reductase catalytic subunit M1 (RRM1) gene silencing on drug resistance of human breast cancer cell line MCF-7/R. We established a paclitaxel-resistant breast cancer MCF-7 cell line (MCF-7/R) by exposing the cells to high-concentration paclitaxel in a short time. Small interfering RNAs (siRNAs) targeting RRM1 were designed to silence RRM1 expression in human breast cancer MCF-7/R cells. MTT assay was used to detect the IC50 values and the sensitivity to paclitaxel in the cells with or without siRNA transfection. The changes in the proliferative activity of MCF7 and MCF-7/R cells following RRM1 gene silencing were evaluated using EdU assay. Flow cytometry was used to analyze the cell apoptosis and cell cycle changes. We assessed the effect of RRM1 gene silencing and paclitaxel on the tumor growth in a nude mouse model bearing subcutaneous xenografts with or without siRNA transfection. We detected significantly higher expressions of RRM1 at both the mRNA and protein levels in the drug-resistant MCF- 7/R cells than in the parental MCF-7 cells (P < 0.01). Transfection with the specific siRNAs significantly reduced the expression of RRM1 in MCF-7/R cells (P < 0.05), which showed a significantly lower IC50 value of paclitaxel than the cells transfected with the negative control siRNA (P < 0.05). RRM1 silencing significantly inhibited the proliferation (P < 0.01) and enhanced the apoptosis-inducing effect of paclitaxel in MCF-7/R cells (P < 0.001); RRM1 silencing also resulted in obviously reduced Akt phosphorylation, suppressed Bcl-2 expression and promoted the expression of p53 protein in MCF-7/R cells. In the tumor-bearing nude mice, the volume of subcutaneously transplanted tumors was significantly smaller in MCF-7/R/siRNA+ PTX group than in the other groups (P < 0.001). RRM1 gene silencing can reverse paclitaxel resistance in human breast cancer cell line MCF-7/R by promoting cell apoptosis.