Human A2780 Research Articles

Rational design, synthesis and biological evaluation 1,2,3-triazole aryl attached (4,6-dimethoxy-1,3,5-triazin-2-yl)thiazole derivatives as anticancer agents

A new library of 1,2,3-triazole aryl-attached (4,6-dimethoxy-1,3,5-triazin-2-yl) thiazoles (11a–j) has been designed and synthesized and its structures were characterized by 1H NMR 13C NMR and mass spectral data. Further, these derivatives (11a-j) were evaluated for their anticancer activity against four human cancer cell lines, including PC3 (prostate cancer), A549 (lung cancer), MCF-7 (breast cancer) and A2780 (ovarian cancer) by employing the MTT method, and the obtained results were compared with etoposide. Among all the examined compounds, 11a, 11b, 11c, 11i and 11j displayed the most promising activity. Particularly, two compounds 11a and 11b possessed good activity.

Abstract 6810: Macrophage secreted CCL9/CCL5 induces CCR1-mediated ovarian cancer metastasis to the omentum in the absence of CCL6

Abstract The tumor-microenvironment (TME) of the omentum plays an important role in providing a pre-metastatic niche for high-grade serous ovarian cancer (HGSC-OvCa) progression. Omentum, a layer of fatty adipose tissue within the peritoneal cavity, is the preferred primary metastatic site for OvCa. Omentum is a well-vascularized fatty tissue containing wide-spread immune clusters (milky spots). Milky spots in the omentum are primarily composed of B cells, T cells, macrophages, NK cells, and dendritic cells. Previous studies from our laboratory have highlighted the importance of CCL6 secreted by omental macrophages as a key chemokine in providing a pre-metastatic niche by activating the CCR1 axis on the cancer cells required for OvCa invasion. We engineered and characterized a CCL6-depleted in vivo mouse model using CRISPR/Cas9 to study the effect on OvCa progression and macrophage biology. We demonstrate that deletion of CCL6 in mice alters characteristics of omental tissue-resident (OM-Macφ) and bone marrow-derived macrophages (BMDM) derived from circulating monocytes. RNA sequencing of OM-Macφ and microarray study of BMDM revealed that CCL6 deletion produces enhanced TGFβ1+CSF-1+CCL5+ expression in OM-Macφ and an increase in TGFβ1+CSF-1+CCL9+ in BMDM. This acts as a rescue mechanism to maintain activation of their shared CCR1 receptor with a pro-M2 stimulation profile. Deep multiplexed imaging of the omentum tissue by CODEX showed that CCL6 deletion alters the architecture of milky spots, with enriched macrophage content and reduced B-cell and T-cell-rich neighborhoods. While our previous studies indicated that CCL6 deletion could potentially have an anti-tumoral effect, we found that it does not significantly affect murine ID8 ovarian cancer tumor burden in both short-term (7 days) and long-term in vivo studies and provides no added survival advantage. We report that CCL9+F4/80+ macrophages accumulate in the omentum post-tumor infiltration in the CCL6-KO model, and neutralizing CCL9 in the CCL6-KO BMDM macrophage-derived conditioned media significantly reduces migration of ID8 cancer cells in vitro. Furthermore, we demonstrate that murine CCL9 and its analog, human CCL15, promote the epithelial-mesenchymal transition (EMT) of mouse ID8 and human A2780 OvCa cells. Moreover, our findings indicate that human CCL15 is a prognostic biomarker for reduced survival in OvCa patient cohorts (Stages I&II and Stage III&IV). Further classification of HGSC-OV patients (n=375) from TCGA-OV based on CCL15 and CCR1-high expression correlated with a higher expression of TGFβ1, CCL5, and a T-cell exhaustion signature in the TME. Taken together, given the apparent redundancy of the chemokine network controlling CCR1 signaling, our findings support the strategy of therapeutically blocking the CCR1 receptor to potentially produce a favorable anti-tumor response in HGSC-OvCa patients. Citation Format: Supreeti Tallapragada, Justine Chan, Venkatesh Krishnan, Oliver Dorigo. Macrophage secreted CCL9/CCL5 induces CCR1-mediated ovarian cancer metastasis to the omentum in the absence of CCL6 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6810.

Methylation of the Selected TP53 Introns in Advanced-Stage Ovarian Carcinomas.

Objective: Advanced-stage ovarian cancer (OC) is among the most fatal female genital tract neoplasms worldwide. Although different genetic mechanisms have been shown to be involved in ovarian carcinogenesis, the role of TP53 introns methylation is still unresolved. We performed methylation analysis of introns 1, 3, and 4 of the TP53 to identify patterns in primary stage III OCs, corresponding metastases, and healthy tissues. Methods: The study involved samples of paraffin-embedded tissues obtained from 80 patients with stage III OCs, who underwent surgery at the Department of Gynecology and Gynecologic Oncology of the Military Institute of Medicine in Warsaw, Poland. Altogether, 40 serous-type G2/3 OCs and 40 endometrioid-type G2/3 OCs were included. From the same patient, metastatic and normal tissues were simultaneously analyzed. As a control group, 80 tissue samples were collected from patients after bariatric operations. Human ovarian cancer A2780 cell line was also investigated. Total genomic DNA was isolated from paraffin-embedded tissue blocks and the methylation analysis was performed by bisulfite DNA conversion, DNA amplification with specific primers, cloning, and DNA sequencing. Results: All of the samples of intron 1 of TP53 were un-methylated in OCs, metastatic tissues, and in healthy tissues from the same patient. Also, no methylation of TP53 intron 1 was detected in cells from the human A2780 ovarian cancer cell line and in all samples from control group. In all samples, introns 3 and 4 of the TP53 were methylated in primary tumors, metastatic tissue, and in healthy tissue from the same patient, in human A2780 ovarian cell line, and in DNA samples from healthy patients. None of the clinicopatholocal features was related to the TP53 introns methylation status. Conclusions: Our data on TP53 introns methylation sheds new light on the mechanism of p53 activity for a better understanding of cancer biology. The study suggests the existence of an additional regulation rule of TP53 activity that involves demethylation-methylation mechanisms. Methylation at introns 3 and 4 may also overall help in protecting TP53 against damage by viral restrictases or viral DNA integration.

New hyaluronan-terpyridine conjugate: Metal complexes and their biological activity

Terpyridine coordination compounds have attracted broad interest concerning their biomedical applications. We devised a terpyridine-hyaluronic acid conjugate and studied its copper(II), iron(II) and zinc(II) metal complexes. We determined the antiproliferative activities of the ligand and the metal complexes in human cell lines A2780 (ovary, adenocarcinoma), A549 (lung, carcinoma), MDA-MB-453 (breast, carcinoma), SKHep1 (liver, carcinoma) and compared them to 4′-Chloro-2,2′:6′,2′’-terpyridine systems. The terpyridine-hyaluronan conjugate showed pharmacologically significant antiproliferative activity in all cell lines tested. The ternary metal complexes of terpyridine-hyaluronan conjugate and 4-chloro-terpyridine showed significant IC50 values (nanomolar range) depending on the cell line. Additionally, Cu2+ and Zn2+ ternary complexes exhibited higher antiproliferative activity than Doxorubicin.

Targeted reversal of multidrug resistance in ovarian cancer cells using exosome‑encapsulated tetramethylpyrazine.

The objective of the present study was to develop exosomes (EXOs) encapsulating tetramethylpyrazine (TMP) for the reversal of drug resistance in ovarian cancer therapy. Human A2780 cells were incubated with TMP for 48h. Purified TMP‑primed EXOs (EXOs‑TMP) were isolated through ultracentrifugation. The developed EXOs‑TMP were characterized using techniques such as transmission electron microscopy, nanoparticle tracking analysis, Fluorescence microscopy and western blotting. Subsequently, MTT, western blotting and flow cytometry assays were performed to evaluate the biological effects in drug‑resistant A2780T cells. The results demonstrated that the incorporation of TMP into EXOs exhibited an anti‑ovarian cancer effect and markedly enhanced the antitumor efficacy of paclitaxel (PTX). Furthermore, it was identified that the ability of EXO‑TMP to reverse cell resistance was associated with the downregulation of multidrug resistance protein1, multidrug resistant‑associated protein1 and glutathione S‑transferase Pi protein expression. Flow cytometry analysis revealed that EXO‑TMP induced apoptosis in drug‑resistant cells and enhanced the apoptotic effect when combined with PTX. EXOs are naturally sourced, exhibit excellent biocompatibility and enable precise drug delivery to target sites, thereby reducing toxic side effects. Overall, EXO‑TMP exhibited direct targeting capabilities towards A2780T cells and effectively reduced their drug resistance. EXOs‑TMP provide a novel and effective drug delivery pathway for reversing drug resistance in ovarian cancer.

Screening of Chenopodium hybridum L. for Anti-ovarian Cancer Compounds Using UPLC-Q-Exactive MS and Partial Least Squares Analysis

Background Extracts of Chenopodium hybridum L. leaves and stems exerted a significant anti-proliferative effect on human A2780 ovarian cancer cells, but C. hybridum active components have not been reported. Materials and Methods Here, a method is described for screening of C. hybridum extracts for potential bioactive components that inhibit A2780 cell proliferation. First, the spectrum–effect relationship between UPLC-Q-Exactive MS chromatograms and C. hybridum extract antiproliferative effect against A2780 cells was established to evaluate extract bioactive components using partial least squares (PLS) analysis. Results The results indicated that the optimal reflux extraction process for preparing C. hybridum extracts with antiproliferative activity involved a suspension of C. hybridum material in 8 volumes of 70% ethanol followed by heating and refluxing twice for 60 min/reflux step and then repeating the extraction and pooling of both the extracts. Chromatographic results revealed five compounds with potential anti-ovarian cancer activities based on inhibition of A2780 cell proliferation: isorhamnetin-3-O-β-D-furanosyl(1→2)-O-[α-L-rhamnpyranosyl(1→6)]-β-D-glucopyranoside, kaempferol-3-O-β-D-glucopyranoside-7-O-α-L-pyranoside, kaempferol-3-O-[α-L-rhamnopyranosyl(1″→2″)]-β-D-galactopyranoside, quercetin-3,7-di-rhamnose, and isorhamnetin-3-acacia disaccharide. Network pharmacological screening revealed nine core cellular targets that potentially interacted with these compounds. Conclusion These results were verified through molecular docking studies that supported the involvement of these compounds in observed C. hybridum A2780 cell antiproliferative effects, thus indicating C. hybridum active components may have value in ovarian cancer treatments.

Design, synthesis, and biological evaluation of simplified tetrahydroisoquinoline analogs.

A series of tetrahydroisoquinoline derivatives were prepared and their antitumor activity was studied against several human carcinoma cell lines, including Ketr3, BEL-7402, BGC-823, KB, HCT-8, MCF-7, HeLa, A2780, A549, and HT-1080. Compound 20, an analog of phthalascidin 650, exhibited good broad-spectrum antitumor activity in vitro. However, compounds 19 and 21, in which the side chains at C-22 are simplified, showed no obvious antitumor activity, indicating that the C-22 side chain of this type of compound has a greater impact on its activity. The difference in the in vivo activity between compound 20 and phthalascidin 650 also shows a significant effect of the substituents on the skeleton structure on the in vivo activity.

Study on the Intervention Mechanism of Cryptotanshinone on Human A2780 Ovarian Cancer Cell Line Using GC-MS-Based Cellular Metabolomics.

Cryptotanshinone (CT), an active component of the traditional Chinese medicine Salvia miltiorrhiza Bunge, exhibits a wide range of biological and pharmacological activities. Although the anticancer activity of CT is well known, the knowledge of its effect on the regulation of cancer cell metabolism is relatively new. The present study investigated the anticancer mechanism of CT in ovarian cancer with a focus on cancer metabolism. CCK8 assays, apoptosis assays, and cell cycle assays were conducted to reveal the growth-suppressive effect of CT on ovarian cancer A2780 cells. To explore the potential underlying mechanisms of CT, the changes in endogenous metabolites in A2780 cells before and after CT intervention were investigated using the gas chromatography-mass spectrometry (GC-MS) approach. A total of 28 important potential biomarkers underwent significant changes, mainly involving aminoacyl-tRNA biosynthesis, energy metabolism, and other pathways. Changes in the ATP and amino acid contents were verified with in vitro and in vivo experiments. Our results indicate that CT may exert an anti-ovarian cancer effect by inhibiting ATP production, promoting the protein catabolic process, and inhibiting protein synthesis, which may lead to cell cycle arrest and apoptosis.

Astragalus Polysaccharide Mediates Immunomodulatory Effects on Crosstalk between Human Peripheral Blood Mononuclear Cells and Ovarian Cancer Cell Line.

Astragalus polysaccharide (APS) is a functional component of Astragalus membranaceus with antitumor and immunomodulatory properties. This study evaluated the effect of APS on the peripheral blood mononuclear cell (PBMC) proliferation, cytokine secretion, and regulatory T cell (Treg) induction in an in vitro coculture model of human PBMCs and A2780 human ovarian cancer cells. PBMC proliferation and Treg frequency were measured by flow cytometry. Cytokine levels were assessed by enzyme-linked immunosorbent assay. APS significantly enhanced the PBMC proliferation, reduced Treg frequency, decreased anti-inflammatory cytokines including interleukin [IL]-10, transforming growth factor beta (TGF-β), and vascular endothelial growth factor-A (VEGF-A), and increased the pro-inflammatory cytokine IL-6. These findings suggest that APS may be an effective immunomodulatory supplement for cancer therapy, particularly for ovarian cancer by enhancing antitumor immune responses.

Newly synthesized benzothiazolyl-1,2,3-triazole derivatives: Intramolecular charge transfer tuning, solvatofluorochromism and antiproliferative properties

A family of 6 new 1-aryl-4-benzothiazolyl-1,2,3-triazoles with CH3, F, CF3, NO2, OCH3 and N(CH3)2 substituents in the R4 of the phenyl ring has been prepared from the respective aldehyde treated with ortho-aminothiophenol. The compounds are characterized in solution using multinuclear NMR spectroscopy and in the solid state by X-ray diffraction. The emission spectra of the compounds with stronger electron-donating groups, OCH3 and N(CH3)2, exhibit a red-shifted band (391 and 515 nm, respectively), the latest showing the largest Stokes shift and the highest quantum yield, which increases with the decrease polarity of the solvent. The excited-state lifetimes of all compounds showed a bi-exponential decay with a short (5–12 ns), according to an admixture of intramolecular charge transfer (ICT) and π-π*, and a long (45–100 ns) component. The calculated structures of their first singlet excited state by TD-DFT in acetonitrile (ACN) solution allows to estimate the change in the dipole moments, which explains the ICT character of their first singlet excited state for the NO2 and N(CH3)2 derivatives. Important to note that compound with the electron-donating N(CH3)2 substituent showed strong solvatofluorochromism (correlated well with the ICT and the dipole moments). Interestingly, some of the compounds are active in human A2780, HeLa and A549 cancer cell lines, exhibiting IC50 values in the low micromolar range, whereas showing low cytotoxicity in healthy CHO cells. Important to note that the N(CH3)2 derivative showed cytoplasmic staining as determined by confocal fluorescent microscopy.

Ruthenium(II) polypyridyl complexes with benzothiophene and benzimidazole derivatives: Synthesis, antitumor activity, solution studies and biospeciation

With the aim to incorporate pharmacophore motifs into the Ru(II)-polypyridyl framework, compounds [Ru(II)(1,10-phenantroline)2(2-(2-pyridyl)benzo[b]thiophene)](CF3SO3)2 (1) and [Ru(II)(1,10-phenantroline)2(2-(2-pyridyl)benzimidazole)](CF3SO3)2 (2) were prepared, characterized and tested for their antitumor potential. The solid-state structure of the compounds was confirmed by single-crystal X-ray diffraction analysis. The solution behavior of both complexes was investigated, namely their solubility, stability, and lipophilicity in physiological mimetic conditions, as well as an eventual uptake by passive diffusion. In vitro anticancer activity of the complexes on ovarian and different colon cancer cells and apoptosis induction by the complexes were studied. A slow transformation process was observed for complex 1 in aqueous solution when exposed to sunlight, while complex 2 undergoes deprotonation (pKa = 7.59). The lipophilicity of this latter complex depends strongly on the pH and ionic strength. In contrast, 1 is rather hydrophilic under various conditions. Complex 1 was highly cytotoxic on Colo-205 human colon (IC50 = 7.87 μM) and A2780 ovarian (IC50 = 2.2 μM) adenocarcinoma cell lines, while 2 displayed moderate anticancer activity (30.9 μM and 18.0 μM, respectively). The complexes induced late apoptosis and necrosis. Only a weak binding of the complexes to human serum albumin, the main transport protein in blood serum, was found. However, a more significant binding to calf thymus DNA was observed in UV–visible titrations and fluorometric dye displacement studies. Detailed analysis of fluorescence lifetime data collected for the latter systems reveals not only the partial intercalation of the complexes, but goes beyond the usual simplified interpretations.

Streptothiazolidine B, a new cytotoxic alkaloid produced by Streptomyces violaceoruber

A new cytotoxic alkaloid, named streptothiazolidine B (1), together with three known compounds (2-4), were isolated from Streptomyces violaceoruber. The structure of the undescribed compound was established using 1D and 2D NMR, and HRESIMS. Streptothiazolidine B was isolated and identified as an amide alkaloid with a unique thiazolidine side chain and its absolute configuration was determined by a combination of NOESY experiment and ECD analysis. Streptothiazolidine B exhibited significant cytotoxic activities against two human tumor cell lines, Li-7 and A2780, with IC50 values of 7.8, and 9.1 μM. Meanwhile, compound 4 showed obvious cytotoxic activities against four human tumor cell lines, THP-1, HT29, Li-7 and A2780, with IC50 values ranging from 3.1 to 10.2 μM.

Abstract 1512: Architectural protein HMGB3 interacts with cisplatin-resistance associated overexpressed protein (CROP/LUC7L3) in human cancer cells and modulates cisplatin-DNA adduct removal

Abstract Cancer treatment and management have improved substantially in recent years resulting in increased overall patient survival. However, upon recurrence, many tumors can eventually stop responding to treatment and become resistant to therapy. Therefore, sensitizing cancer cells to therapy is an important approach to increase overall patient survival and improve their quality of life. We have previously shown that targeting the chromatin associated architectural protein High Mobility Group Box 3 (HMGB3) can sensitize cisplatin resistant human ovarian cancer cells to cisplatin. HMGB proteins are non-histone chromatin proteins, which bind to distorted DNA structures with higher affinity compared to double-stranded DNA. HMGB proteins have been shown to play important roles in DNA metabolism, such as replication, transcription and repair. We found that the sensitization of cisplatin resistant human A2780/CP70 ovarian cancer cells by siRNA-mediated targeting of HMGB3 is achieved, at least in part, by significantly lowering the expression of the DNA damage response (DDR) proteins ATR and CHK1. In addition, depletion of HMGB3 repressed global transcription by ~50% in human osteosarcoma (U2OS) cells. Here we report that GFP-tagged HMGB3 protein forms distinct foci in human A2780 ovarian cancer cells upon cisplatin treatment. Further, depleting HMGB3 in cisplatin resistant A2780/CP70 cells reduces the rate of cisplatin-DNA adduct removal, suggesting a possible role in chemoresistance. In addition, mass spectrometry studies reveal that HMGB3 interacts with the cisplatin resistance-associated overexpressed protein (CROP, also known as LUC7L3 ), following treatment with the DNA interstrand crosslink-inducing agent, psoralen (plus UVA irradiation). LUC7L3 is associated with the spliceosome and is overexpressed in A2780/CP70 cells compared to A2780 ovarian cancer cells. SiRNA-mediated depletion of HMGB3 reduces the transcription levels of LUC7L3. Further, siRNA-mediated depletion of LUC7L3 reduced the clone forming capacity of U2OS cells and ovarian cancer cells. Analysis of cancer patient databases revealed that LUC7L3 mRNA is overexpressed in many cancer types including ovarian cancer. Further analysis showed that ovarian cancer, gastric cancer, liver cancer and lung cancer patients overexpressing LUC7L3 have a significantly reduced overall survival than those with lower levels. Taken together, these data indicate that in addition to the architectural roles of HMGB3, which is important for DNA metabolism, transcriptional modulation of DDR proteins through interaction with the spliceosome may be another way that HMGB3 modulates chemotherapeutic DNA damage processing in human cancer cells. Citation Format: Anirban Mukherjee, Lorraine-Rana Benhamou, Karen M. Vasquez. Architectural protein HMGB3 interacts with cisplatin-resistance associated overexpressed protein (CROP/LUC7L3) in human cancer cells and modulates cisplatin-DNA adduct removal [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1512.

Synthesis and Anticancer Assessment of Various Amide Derivatives of Imidazo[2,1-b]Oxazoles as Anticancer Agents

A new series of amide derivatives (9a–j) have been synthesized and their chemical structures characterized by the 1HNMR, 13CNMR, and mass spectroscopic techniques. The synthesized amide derivatives (9a–j) were evaluated for their in vitro anti-proliferative activity against four human cancer cell lines, including PC3 (prostate cancer), A549 (lung cancer), MCF7 (breast cancer), and A2780 (ovarian cancer) cells by employing the MTT assay. Results were expressed as IC50 values and were compared with the clinical drug, etoposide, used as a positive control. Among the synthesized amide derivatives, five compounds 9a, 9b, 9c, 9d, and 9g revealed potent anti-proliferative potential toward all cell lines. Primarily, two compounds 9a and 9b possessed the most promising anticancer activities.

Effect of cysteine thiols on the catalytic and anticancer activity of Ru(II) sulfonyl-ethylenediamine complexes.

We have synthesized a series of novel substituted sulfonyl ethylenediamine (en) RuII arene complexes 1-8 of [(η6-arene)Ru(R1-SO2-EnBz)X], where the arene is benzene, HO(CH2)2O-phenyl or biphenyl (biph), X = Cl or I, and R1 is phenyl, 4-Me-phenyl, 4-NO2-phenyl or dansyl. The 'piano-stool' structure of complex 3, [(η6-biph)Ru(4-Me-phenyl-SO2-EnBz)I], was confirmed by X-ray crystallography. The values of their aqua adducts were determined to be high (9.1 to 9.7). Complexes 1-8 have antiproliferative activity against human A2780 ovarian, and A549 lung cancer cells with IC50 values ranging from 4.1 to >50 μM, although, remarkably, complex 7 [(η6-biph)Ru(phenyl-SO2-EnBz)Cl] was inactive towards A2780 cells, but as potent as the clinical drug cisplatin towards A549 cells. All these complexes also showed catalytic activity in transfer hydrogenation (TH) of NAD+ to NADH with sodium formate as hydride donor, with TOFs in the range of 2.5-9.7 h-1. The complexes reacted rapidly with the thiols glutathione (GSH) and N-acetyl-L-cysteine (NAC), forming dinuclear bridged complexes [(η6-biph)2Ru2(GS)3]2- or [(η6-biph)2Ru2(NAC-H)3]2-, with the liberation of the diamine ligand which was detected by LC-MS. In addition, the switching on of fluorescence for complex 8 in aqueous solution confirmed release of the chelated DsEnBz ligand in reactions with these thiols. Reactions with GSH hampered the catalytic TH of NAD+ to NADH due to the decomposition of the complexes. Co-administration to cells of complex 2 [(η6-biph)Ru(4-Me-phenyl-SO2-EnBz)Cl] with L-buthionine sulfoximine (L-BSO), an inhibitor of GSH synthesis, partially restored the anticancer activity towards A2780 ovarian cancer cells. Complex 2 caused a concentration-dependent G1 phase cell cycle arrest, and induced a significant level of reactive oxygen species (ROS) in A2780 human ovarian cancer cells. The amount of induced ROS decreased with increase in GSH concentration, perhaps due to the formation of the dinuclear Ru-SG complex.

LINC00035 Transcriptional Regulation of SLC16A3 via CEBPB Affects Glycolysis and Cell Apoptosis in Ovarian Cancer.

Objective Ovarian cancer (OC) represents the most lethal gynecologic malignancy globally. Over the decades, lncRNAs have been considered as study focuses due to their genome-wide expression through multiple mechanisms in which regulation of target gene transcription through interaction with transcription factors or epigenetic proteins is proven. In the present work, we focus on the functional role of LINC00035 in OC and its regulation mechanism on gene expression. Methods We collected OC tissues and adjacent tumor-free tissues surgically resected from 67 OC patients. Cultured human OC cell lines SKOV3 and A2780 were assayed for their viability, migration, invasion, apoptosis in vitro using CCK-8 assays, transwell assays, and flow cytometric analysis. OC cell tumorigenesis in vivo was evaluated by mouse xenograft experiments. Glycolysis was evaluated by glucose uptake, lactate release, and ATP production assays. Luciferase activity assay, RNA immunoprecipitation (RIP), and RNA pull-down were performed to confirm the interactions among LINC00035, CEBPB, and SLC16A3. Results LINC00035 was upregulated in OC tissues. LINC00035 knockdown was shown to repress SKOV3 and A2780 cell viability, migration, invasion, induce their apoptosis, and reduce glucose uptake, lactate release, and ATP production. LINC00035 could recruit CEBPB into the SLC16A3 promoter region, thus increasing the SLC16A3 transcription. SLC16A3 was upregulated in OC tissues. SLC16A3 knockdown exerted similar effects on SKOV3 and A2780 cells as LINC00035 knockdown. Rescue experiments found SLC16A3 overexpression resisting to LINC00035 knockdown on SKOV3 and A2780 cell viability, migration, invasion, apoptosis, glucose uptake, lactate release, and ATP production. Results also showed LINC00035 knockdown could inhibit OC cell tumorigenesis in vivo. Conclusion The study reveals that LINC00035 promotes OC progression by regulating glycolysis and cell apoptosis through CEBPB-mediated transcriptional promotion of SLC16A3.

DNA-Intercalative Platinum Anticancer Complexes Photoactivated by Visible Light.

Photoactivatable agents offer the prospect of highly selective cancer therapy with low side effects and novel mechanisms of action that can combat current drug resistance. 1,8‐Naphthalimides with their extended π system can behave as light‐harvesting groups, fluorescent probes and DNA intercalators. We conjugated N‐(carboxymethyl)‐1,8‐naphthalimide (gly‐R‐Nap) with an R substituent on the naphthyl group to photoactive diazido PtIV complexes to form t,t,t‐[Pt(py)2(N3)2(OH)(gly‐R‐Nap)], R=H (1), 3‐NO2 (2) or 4‐NMe2 (3). They show enhanced photo‐oxidation, cellular accumulation and promising photo‐cytotoxicity in human A2780 ovarian, A549 lung and PC3 prostate cancer cells with visible light activation, and low dark cytotoxicity. Complexes 1 and 2 exhibit pre‐intercalation into DNA, resulting in enhanced photo‐induced DNA crosslinking. Complex 3 has a red‐shifted absorption band at 450 nm, allowing photoactivation and photo‐cytotoxicity with green light.

Ultrasound-Assisted Water Extraction of Mastocarpus stellatus Carrageenan with Adequate Mechanical and Antiproliferative Properties.

Ultrasound-assisted water extraction was optimized to recover gelling biopolymers and antioxidant compounds from Mastocarpus stellatus. A set of experiments following a Box–Behnken design was proposed to study the influence of extraction time, solid liquid ratio, and ultrasound amplitude on the yield, sulfate content, and thermo-rheological properties (viscoelasticity and gelling temperature) of the carrageenan fraction, as well as the composition (protein and phenolic content) and antiradical capacity of the soluble extracts. Operating at 80 °C and 80 kHz, the models predicted a compromise optimum extraction conditions at ~35 min, solid liquid ratio of ~2 g/100 g, and ultrasound amplitude of ~79%. Under these conditions, 40.3% carrageenan yield was attained and this product presented 46% sulfate and good mechanical properties, a viscoelastic modulus of 741.4 Pa, with the lowest gelling temperatures of 39.4 °C. The carrageenans also exhibited promising antiproliferative properties on selected human cancer cellular lines, A-549, A-2780, HeLa 229, and HT-29 with EC50 under 51.9 μg/mL. The dried soluble extract contained 20.4 mg protein/g, 11.3 mg gallic acid eq/g, and the antiradical potency was equivalent to 59 mg Trolox/g.

Atrazine Promoted Epithelial Ovarian Cancer Cells Proliferation and Metastasis by Inducing Low Dose Reactive Oxygen Species (ROS).

Background: Atrazine (ATZ) is a triazine herbicide that is widely used in agriculture and has been detected in surface and underground water. Recently, laboratory and epidemiological research have found that the bioaccumulation of ATZ in the environment leads to biotoxicity in the human immune and endocrine systems and results in tumor development.Objective: To investigate the effects of ATZ exposure on epithelial ovarian cancer (EOC) cells and elucidate the potential mechanisms governing these effects.Materials and Methods: The human EOC cell lines Skov3 and A2780 were used in this study to explore the effects and mechanisms of ATZ exposure on EOC. The mouse embryonic osteoblastic precursor MC3T3-E1 cells served as the control cells to determine the effects of ATZ on cancer cell lines. After exposure to ATZ, the MTT assay, flow cytometry, the colony formation assay, immunohistochemical staining, the cell scratch assay, and the Transwell assay were used to evaluate the proliferative activity, invasion, and migration capabilities of EOC cell lines. Moreover, flow cytometry was also applied to detect the level of reactive oxygen species (ROS) in these two EOC cell lines, as well as the MC3T3-E1 cells. To further illustrate the underlying mechanisms governing the effect of ATZ on EOC, real-time PCR and Western blotting were employed to assess the transcription and the expression level of Stat3 signaling pathway-related genes in Skov3 and MC3T3-E1 cells.Results: The results showed that following ATZ treatment, the cell proliferation, migration, and invasion potencies of Skov3 and A2780 cells were increased compared to those of the control group. Meanwhile, the ROS levels of EOC and MC3T3-E1 cells were notably elevated after ATZ treatment. In Skov3 cells, the expression levels of p53 and p21 were downregulated, while those of Cyclin E, vascular endothelial growth factor (VEGF), matrix metallopeptidase 2 (MMP2), MMP9, signal transducers and activators of transcription 3 (Stat3), and p-Stat3 were upregulated by ATZ treatment. In MC3T3-E1 cells, however, ATZ treatment did not affect the level of p53/p21 mRNA compared to the control groups. Moreover, there was no significant change in the expression levels of Stat3 and p-Stat3 in MC3T3-E1 cells exposed to ATZ. This phenomenon was observed while the proliferation rate was enhanced in MC3T3-E1 cells by ATZ.Conclusions: The results of this study suggest that ATZ effectively promotes the proliferation and metastasis of EOC cells through the Stat3 signaling pathway by inducing low levels of ROS. Additionally, although ATZ might also induce proliferative potential in normal cells, the mechanisms governing its effects in these cells might be different from those in EOC cells.

Elevated expression of Tweety homologue 3 predicts poor clinical outcomes in ovarian cancer.

Objective: To define the alteration of tweety homolog (TTYH) expression in patients with ovarian carcinoma (OC) and its correlation to prognosis.Methods: Kaplan-Meier (KM) plotter was used to evaluate the association between TTYHs expression and clinical outcomes of OC patients. The distribution of 20-year overall survival (OS) and progression-free survival (PFS) was estimated using KM survival plots. The mRNA expression of TTYHs in OC and normal ovarian tissues was confirmed by the Oncomine database. Then, using immunohistochemistry assay, the expression of TTYH1 and TTYH3 proteins in serous OC and normal ovarian tissues was detected. In addition, the protein and mRNA levels of TTYH1 and TTYH3 in human OC cell lines ES-2, A2780 and SKOV3 and normal ovarian epithelial cell lines IOSE80 were assessed by western blotting and real-time quantitative polymerase chain reaction (qRT-PCR).Results: TTYH1 possessed meaningful significance in predicting better prognosis in the serous, advanced stage, and well-differentiated OC patients, while TTYH3 expression predicted worse prognosis in serous, late-stage, and poorly differentiated OC patients. High expression of TTYH1 displayed an association with favorable PFS in OC patients with TP53 mutation. However, enhanced TTYH3 was related to an adverse clinical outcome in TP53-mutated OC patients. In addition, TTYH1 was related to a better clinical outcome in OC patients with platinums-based chemotherapy, but only indicated improved overall survival in OC patients who received taxol or platin + taxol chemotherapy. The up-regulated expression of TTYH3 predicted worse survival in OC patients receiving platin, taxol, or platin + taxol chemotherapy regimen. The levels of TTYH3 mRNA and protein were higher in OC cells and tissues when compared to normal ovarian cells and tissues.Conclusions: TTYH3 was a potential predictor for poor clinical outcome in OC patients, particularly in patients with serous, late-stage, poorly differentiated, TP53-mutation or the patients treated with chemotherapy regimens (platin, taxol, or platin + taxol).