Ductal Breast Epithelial Tumor Cell Research Articles

Platinum(II) Complexes with Bulky Disubstitute Triazolopyrimidines as Promising Materials for Anticancer Agents.

Herein, we present dicarboxylate platinum(II) complexes of the general formula [Pt(mal)(DMSO)(L)] and [Pt(CBDC)(DMSO)(L)], where L is dbtp 5,7-ditertbutyl-1,2,4-triazolo[1,5-a]pyrimidine) or ibmtp (7-isobutyl-5-methyl-1,2,4- triazolo[1,5-a]pyrimidine), as prospective prodrugs. The platinum(II) complexes were synthesized in a one-pot reaction between cis-[PtCl2(DMSO)2], silver malonate or silver cyclobutane-1,1-dicarboxylate and triazolopyrimidines. All platinum(II) compounds were characterized by FT-IR, and 1H, 13C, 15N and 195Pt NMR; and their square planar geometries with one monodentate N(3)-bonded 5,7-disubstituted-1,2,4-triazolo[1,5-a]pyrimidine, one S-bonded molecule of dimethyl sulfoxide and one O,O-chelating malonato (1, 2) or O,O-chelating cyclobutane-1,1-dicarboxylato (3, 4) was determined. Additionally, [Pt(CBDC)(dbtp)(DMSO)] (3) exhibited (i) substantial in vitro cytotoxicity against the lung adenocarcinoma epithelial cell line (A549) (IC50 = 5.00 µM) and the cisplatin-resistant human ductal breast epithelial tumor cell line (T47D) (IC50 = 6.60 µM); and (ii) definitely exhibited low toxicity against normal murine embryonic fibroblast cells (BALB/3T3).

Abstract 497: Higher nucleolar index of nucleolin as an indicator of aberrant cellular DNA damage response (DDR)

Abstract Nucleoli provide a survival advantage for various tumor types, exemplified in prostate neoplasia, leukemia, lung, and breast carcinomas. Unregulated expression and increased nucleolar localization of many stress-responsive factors often correlate with hyper-proliferative status. Therefore, nucleolar changes in cancer cells are the first cytological indicators that can serve as an index for diagnostic and prognostic purposes. In this study, we examine an abundant nucleolar stress factor, nucleolin (NCL) for its sub-nuclear localization under normal and DNA damage conditions. NCL is a multifunctional RNA-binding phosphoprotein that plays critical role/s in many cellular processes including regulation of gene expression during normal cell cycle as well as in the cellular DNA damage response (DDR). NCL-mediated DDR involves changes in its phosphorylation status, sub-nuclear translocation and a multi-level regulation of p53-checkpoint signaling. However, the role/s of p53 status and the signaling pathways triggered during NCL-mediated stress response remain elusive, and forms the focus of this study. Taking advantage of two human ductal breast epithelial tumor cell lines with different genetic backgrounds (wild type, wt-p53 and mutant, mut-p53 L194F), we demonstrate that NCL is predominantly nucleolar in unstressed conditions irrespective of the p53 status. Upon genotoxic stress, however, wt-p53 is required for NCL to mobilize from nucleoli to the nucleoplasmic region. We observed a significant increase in nucleoplasmic index for NCL that is wt-p53 dependent and specific to the type of DNA damage. In contrast, in the presence of mut-p53, NCL failed to translocate even under various DNA damage conditions, resulting in a higher nucleolar index. Interestingly, another nucleolar factor, nucleophosmin (NPM) remained unaltered upon stress conditions even in the presence of wt-p53. This study corroborates the previous research that suggests NCL translocation requires wt-p53 and provides new insights into the role of p53-mutations (e.g. the zinc finger mutation in the central DNA binding region-L194F) in NCL-mediated DDR. Further, we also assessed the role of NCL and p53 (wt vs. mut) in regulating stress signaling (ATM/ATR and p38-kinase) pathways and gene expression in apoptosis. Together, our study elaborates a novel approach that correlates NCL sub-nuclear localization, downstream signaling pathways and gene expression during the cellular DDR. Citation Format: Iqra Nadeem, Amna Aslam, Jingyuan Wang, Anna Kozlova, Danielle Gordon, Rumsha Javed, Ruchama Steinberg, Rachele Dolce Rameau, Xinyin Jiang, Anjana D. Saxena. Higher nucleolar index of nucleolin as an indicator of aberrant cellular DNA damage response (DDR) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 497. doi:10.1158/1538-7445.AM2017-497

Design and Synthesis of Pyrrolo[2,1-a]Isoquinoline-Based Derivatives as New Cytotoxic Agents.

A new series of anti-cancer agents based on 1,2-diaryl-5,6-dihydropyrrolo[2,1-a]isoquinoline scaffold containing N,N-diethylamino-ethoxy, piperidinyl-ethoxy or morpholinyl-ethoxy group at the para position of the C-2 phenyl ring were synthesized and their cytotoxic activities were assessed against several human cancer cell lines including MCF-7 (ER positive breast cancer cell), MDA-MB231 (ER-negative breast cancer cell), T47D (Human ductal breast epithelial tumor cell line), A549 (adenocarcinomic human alveolar basal epithelial cells), and Hela (human cervix adenocarcinoma cells) using MTT assay. Based on results, compounds, 1-(4-(2-(piperidin-1-yl)ethoxy)phenyl)-5,6-dihydro-8,9-dimethoxy-2-phenylpyrrolo[2,1-a]isoquinoline (6a) and 2-(4-(5,6-dihydro-8,9-dimethoxy-2-phenylpyrrolo[2,1-a] isoquinolin-1-yl)phenoxy)-N,N-diethylethanamine (6c) were the most potent cytotoxic compounds and more toxic than the reference compound against T47D cell line, while all the compounds had satisfactory activity against HeLa cell line with mean IC50 values ranging from 1.93 to 33.84 µM.

Synthesis and anticancer and lipophilic properties of 10-dialkylaminobutynyl derivatives of 1,8- and 2,7-diazaphenothiazines

New derivatives of two isomeric types of azaphenothiazines, 1,8- and 2,7-diazaphenothiazine, containing the triple bond substituents and additionally tertiary cyclic and acyclic amine groups, were synthesized and tested for their anticancer activity. The compounds exhibited differential inhibitory activities. Better results were obtained when the acetylenic group was transformed via the Mannich reaction to the dialkylaminobutynyl groups. The most active was 2,7-diazaphenothiazine with the N-methylpiperazine-2-butynyl substituent against the human ductal breast epithelial tumor cell line T47D, more potent than cisplatin. The 2,7-diazaphenothiazine system turned out to be more active than isomeric 1,8-diaza one. For the most active compound, the expression of TP53, CDKN1A, BCL-2 and BAX genes was detected by the RT-QPCR method. The gene expression ratio BACL-2/BAX suggests the mitochondrial apoptosis in T47D cells. The synthesis makes possible to obtain many new bioactive phenothiazines with the dialkylaminoalkynyl substituents inserting various tertiary cyclic and acyclic amine moieties to the substituents.

DNA scission inhibition, antioxidant, and antiproliferative activities of water chestnut (Trapa natans) extracted in different solvents

This investigation deals with the study of polyphenolic, antioxidant, and antiproliferative activities of water chestnut extracted in different solvents. The antioxidant activities of the extract in different solvents were the maximum in methanol, followed by ethanol and aqueous solution and were found to be 39.8%, 34.1% and 32.6% from DPPH assay, and 45.64%, 35.08% and 30.52% from lipid peroxidation assay, respectively, whereas the reducing power inhibition assay showed the maximum antioxidant potential in the aqueous extract of the sample, followed by methanolic and ethanolic extracts. The polyphenolic content of the water chestnut was found to be more in methanol than in aqueous and ethanolic extracts. The water chestnut extract also showed the antiproliferative potential when tested against the human colon cancer cell line (Colo-205), human ductal breast epithelial tumor cell line (T47D), and human breast adenocarcinoma cell line (MCF7). It also showed some protection from DNA damage induced by hydroxyl radicals in calf thymus DNA.

Design, Synthesis and Cytotoxicity Evaluationof New 1,2-diaryl-4, 5, 6, 7-Tetrahydro-1H-benzo[d] Imidazolesas Tubulin Inhibitors.

A new series of 1,2-diaryl-4,5,6,7-tetrahydro-1H-benzo[d]imidazoles, possessing trimethoxyphenyl pharmacophore, were synthesized to evaluate their biological activities as tubulin inhibitors. Cytotoxic activity of the synthesized compounds 7a-f was assessed against several human cancer cell lines, including MCF-7 (breast cancer cell), HEPG2 (liver hepatocellular cells), A549 (adenocarcinomic human alveolar basal epithelial cells), T47D (Human ductal breast epithelial tumor cell line) and fibroblast. According to our results, HEPG2 seems to be the most sensitive, while MCF7 was the most resistant cell line to the compounds. All the compounds expect 7b, possessed satisfactory activity against HEPG2 with mean IC50 values ranging from 15.60 to 43.81 µM.

Cytotoxic malonate platinum(II) complexes with 1,2,4-triazolo[1,5-a]pyrimidine derivatives: Structural characterization and mechanism of the suppression of tumor cell growth

A series of malonate (mal) platinum(II) complexes of the general formula [Pt(mal)(L)2], where L=5,7-dimethyl-1,2,4-triazolo[1,5-a]pyrimidine (dmtp) (1), 7-isobutyl-5-methyl-1,2,4-triazolo[1,5-a]pyrimidine (ibmtp) (2) or 5,7-ditertbutyl-1,2,4-triazolo[1,5-a]pyrimidine (dbtp) (3), has been prepared and characterized using multinuclear (1H, 13C, 15N, 195Pt) NMR, IR and electrospray ionization mass spectrometry (ESIMS). Furthermore, the crystal structures of [Pt(mal)(dmtp)2]∙4H2O (1a) and [Pt(mal)(dbtp)2]∙CHCl3 (3a) have been determined using single-crystal X-ray diffraction. The spectroscopic characterization unambiguously confirmed the square-planar geometry of Pt(II) with two monodentate N3-bonded 5,7-disubstituted-1,2,4-triazolo[1,5-a]pyrimidines and one O-chelating malonate. The antiproliferative activities of the compounds against the human cell lines T47D (cisplatin-resistant human ductal breast epithelial tumor cell line) and A549 (lung adenocarcinoma epithelial cell line) and the mouse cell line 4T1 (mouse breast tumor model) were assessed using an in vitro screening assay. Compounds (2) and (3) exhibited substantial antigrowth properties against T47D cells, whereas only (3) exhibited an IC50 value that was lower than cisplatin and carboplatin against the 4T1 cell line. Additionally, compounds (2, 3) are capable of arresting the cell cycle of A549 cells at the G0/G1 phase, whereas cisplatin and carboplatin arrested the cells at the G2/M phase, indicating differences in the mechanism of the suppression of tumor cell growth. Finally, in the quest for low toxicity platinum drugs, the in vitro antiproliferative activity against normal mouse fibroblast cells (BALB/3T3) was evaluated. The inhibition of BALB/3T3 cell proliferation by the evaluated Pt(II) complexes increased in the order (1)<(2)<<carboplatin<<(3)<cisplatin.

Design, Synthesis and Cytotoxicity Evaluation of New 2-Aryl-5, 6-Dihydropyrrolo[2, 1-a]Isoquinoline Derivatives as Topoisomerase Inhibitors.

Two set of 2-aryl-5, 6-dihydropyrrolo [2,1-a] isoquinolines were designed and synthesized to evaluate their biological activities as topoisomerase inhibitors. Cytotoxic activity of the synthesized compounds 4a-e and 7a-d was assessed against several human cancer cell lines, including MCF-7 (breast cancer cell), HepG2 (liver hepatocellular cells), A549 (adenocarcinomic human alveolar basal epithelial cells), T47D (Human ductal breast epithelial tumor cell line) and Hela (Human cervix cancer). According to our results, HepG2 seems to be the most sensitive cell line for these compounds with mean IC50 values ranging from 4.25 to 70.05 μM. Our results indicated that compound 7b exhibited the best potency against the tested cell lines. These results also suggest that pyrroloisoquinoline moiety constitutes a suitable scaffold to design new anti-proliferative agents.

Anticancer and free radical scavenging potency of Catharanthus roseus, Dendrophthoe petandra, Piper betle and Curcuma mangga extracts in breast cancer cell lines

This research was conducted to identify the anticancer and antioxidant activity of Catharanthus roseus [L] G.Don, Dendrophtoe petandra L, Piper betle L and Curcuma mangga Val aqueous extracts in T47D human ductal breast epithelial tumor cell line. The anticancer potency was determined via the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay while the apoptotic activity was determined with Sub-G1 flow cytometric analysis. The antioxidant activity was determined by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. C.roseus was able to reduce T47D cell proliferation with a median inhibition concentration (IC50) of 2.8%; D.petandra 1.2%; P.betle 2.8%; and C.mangga 74.8%. The apoptotic analysis result showed that C.roseus induced apoptosis for 37.67%; D.petandra 24.03%; P.betle 9.45%; and C.mangga 0.41%. Meanwhile doxorubicin at 10 μg/ml induced apoptosis for 36.06%. The highest DPPH scavenging activity was recorded for P.betle extract as to be 83%; D.petandra 75.11%; C.roseus 71.87%; and the lowest for C.mangga as 38.45%. C.moseus and D.petandra aqueous extracts presented the highest anticancer activity by means of cell proliferation inhibition and inducing apoptosis. P.betle extract represented the strongest antioxidant activity. C.mangga extract exhibited no anticancer and only low antioxidant activity.

Molecular Mechanism of Hepcidin-Mediated Ferroportin Internalization Requires Ferroportin Lysines, Not Tyrosines or JAK-STAT

Ferroportin is the primary means of cellular iron efflux and a key component of iron metabolism. Hepcidin regulates Fpn activity by inducing its internalization and degradation. The mechanism of internalization is reported to require JAK2 activation, phosphorylation of Fpn tyrosine residues 302 and 303, and initiation of transcription through STAT3 phosphorylation. These findings suggest Fpn may be a target for therapeutic intervention through JAK2 modulation. To evaluate the proposed mechanism, Fpn internalization was assessed using several techniques combined with reagents that specifically recognized cell-surface Fpn. Invitro results demonstrated that Hepc-induced Fpn internalization did not require JAK2 or phosphorylation of Fpn residues 302 and 303, nor did it induce JAK-STAT signaling. Invivo, inhibition of JAK2 had no effect on Hepc-induced hypoferremia. However, internalization was delayed by mutation of two Fpn lysine residues that may be targets of ubiquitination.

Abstract 1182: Targeting p53 to mitochondrial outer membrane using the Bcl-XL signal causes p53 specific apoptosis

Abstract The tumor suppressor p53 can cause cell cycle arrest, DNA repair, and apoptosis via its function as a nuclear transcription factor. However, p53 also triggers apoptosis through a mitochondrial pathway. Mitochondrial p53 binds to anti-apoptotic (Bcl-2, Bcl-XL) and pro-apoptotic (Bak, Bax) proteins localized at the mitochondrial outer membrane (MOM), causing MOM permeabilization that eventually results in apoptosis. The purpose of this work was to develop an optimal version of p53 engineered with a mitochondrial targeting signal (MTS) to trigger rapid apoptosis via the intrinsic pathway. To further investigate mitochondrial activity of p53, we targeted p53 to specific mitochondrial compartments. EGFP-p53 was fused to different mitochondrial targeting signals (MTSs) directing it to the mitochondrial outer membrane (“XL-MTS” from Bcl-XL; “TOM-MTS” from TOM20), inner membrane (“CCO-MTS” from cytochrome c oxidase) or matrix (“OTC-MTS” from ornithine transcarbamylase). The apoptotic mechanism for each construct was further studied using pifithrin-μ (which inhibits p53-mediated transcriptional activity), pifithrin-μ (which reduces binding of p53 to Bcl-2 and Bcl-XL), and by over-expressing the anti-apoptotic Bcl-XL. Apoptosis assays (caspase-9, TUNEL, Annexin-V, 7-AAD) were performed in T47D cells (human ductal breast epithelial tumor cell line). While all MTS-EGFP-p53 constructs showed apoptotic potential, only EGFP-p53-XL had the most significant apoptotic potential compared to its own EGFP-XL control. EGFP tagged to OTC, CCO and TOM tend to interfere with sensitive mitochondrial function versus EGFP-XL, which demonstrated minimal toxicity. Previous work in the literature found that targeting p53 to the mitochondrial matrix causes apoptosis. However, our work indicates targeting even non-toxic proteins (EGFP) to the matrix might cause some level of toxicity due to the MTS by itself. Further, we inquired if the apoptotic response was due to the Bcl-XL dependent pathway. CCO-EGFP-p53 initiated apoptosis mainly through the transcriptional activity of p53 (rescued by pifithrin-μ). TOM-EGFP-p53 is thought to cause apoptosis by activating the pro-apoptotic Bak (rescued by over-expressing Bcl-XL). OTC-EGFP-p53 resulted in apoptosis through binding to Bcl-XL (rescued by pifithrin-μ and over-expressing Bcl-XL) and showed additional apoptotic potential through transcriptional activity of p53 (rescued by pifithrin-μ). In addition, EGFP-p53-XL was the most specific to the p53/Bcl-XL mitochondrial pathway (rescued by pifithrin-μ and over-expressing Bcl-XL). Taken together our data highlights that targeting p53 to Bcl-XL via the XL MTS triggers a rapid apoptotic response through an interaction of p53 with anti-apoptotic Bcl-XL. Due to the ubiquitous role of p53, this gene therapy approach could be used in many different types of cancers. Acknowledgements: NIH CA151847 Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1182. doi:1538-7445.AM2012-1182

COX-2 and survivin reduction may play a role in berberine-induced apoptosis in human ductal breast epithelial tumor cell line

Berberine is an isoquinoline alkaloid that has several pharmacological effects such as antiinflammatory, antimicrobial, apoptosis-inducing and anticancer effects. It has been illustrated that the antiinflammatory effect is mediated by suppressing the nuclear factor-kappa B (NF-κB) that activates expression of some antiinflammatory and antiapoptotic proteins including cyclooxygenase-2(COX-2), inducible nitric oxide synthase (iNOS) and survivin; therefore, berberine may induce apoptosis by reducing antiinflammatory and antiapoptotic agents, which suggest the relationship between antiinflammatory and apoptosis pathways. For further illustration of the mechanism of berberine action, the human ductal breast epithelial tumor cell line (T47D cell line) was treated with different concentrations of berberine (25-100 μM/ml). Berberine in 50 μM/ml had the most reducing effect on cell viability and inducing of apoptosis. The level of COX-2, iNOS and survivin proteins decreased in berberine-treated cells; however, treatment of the cells with aspirin and aminoguanidine (AG), COX-2 and iNOS inhibitors, respectively, showed that despite the cell growth-reducing effect of aspirin, AG did not have a significant effect on cell viability. On the other hand, with the attention to reduction in survivin protein level in berberine-treated cells, the results suggest that the apoptotic effect of berberine may be mediated by reduction in both of the COX-2 and survivin in T47D cell line, while the iNOS does not play any effective role in berberine-induced apoptosis.