Chosen Cell Lines Research Articles

Rough edges of reduced graphene oxide (rGO) sheets elicit anticancerous activities: An in vitro study

In this study, reduced graphene oxide nanosheets (rGO) were synthesized using a chemical bottom-up technique of modified Hummer’s method. The as-synthesized graphene sheets were characterized optically by Fourier Transform Infra Red spectroscopy (FTIR) and Raman spectroscopy. The morphological characterizations were performed using Field-Emission Scanning Electron Microscopy (FE-SEM), High-Resolution Transmission Electron Microscopy (HR-TEM), Atomic Force Microscopy (AFM) and Confocal Microscopy, respectively. It could be evident from the images that wrinkles and surface corrugation are clearly visible from the morphological analysis. The biomedical applications of rGO nanosheets have been studied in the cancer cell lines of A375 and A549 cells at the in-vitro conditions. MTT assay and live dead assay results clearly indicated that rGO exhibited robust anticancerous activities against the chosen cell lines. Although, it could not elicit any antimicrobial activity at the selected doses. Understanding the primary mechanism and the underlying phenomenon involving the interaction of the rGO nanosheets with the cell systems reveals the potentialities of the nanosheet in designing future-based anticancerous therapeutics at the nanoscale level.

Novel green synthesis of zinc oxide nanoparticles using Salvia rosmarinus extract for treatment of human lung cancer

Abstract A green and low-cost approach was run to synthesize zinc nanoparticles (NPs) using rosemary extract. The NPs were identified by various methods, i.e., ultraviolet-visible and Fourier transform infrared spectroscopy, FE-scanning electron microscope imaging, X-ray diffraction, and energy-dispersive X-ray analysis. The radical scavenging activity and MTT assays were used to evaluate the biological activity of ZnO-NPs@Rosemary. The results revealed a spherical shape for ZnO-NPs@Rosemary with a crystal size of 30.74 nm. ZnO-NPs@Rosemary could scavenge the free radicals of DPPH with an IC50 of 87.62 ± 0.47 μg/mL. An MTT assay was run to investigate the anti-cancer activity of ZnO-NPs@Rosemary against PC-14, LC-2/ad, and HLC-1 as the selected lung cancer cell lines. The highest sensitivity of NPs was found against PC-14 with IC50 of 178.84 ± 2.13. A dose-dependent activity was observed for ZnO-NPs@Rosemary against the chosen cell lines. The outcomes of the present study revealed an acceptable anti-lung cancer activity of ZnO-NPs@Rosemary.

Chitosan-Based Nanocarriers for Delivery of Remdesivir

Stable multicomponent capsules for the delivery of remdesivir (Veklury®) are produced through subsequent electrostatic adsorption of oppositely charged components on oil emulsion droplets. For the first time, the encapsulation and release of the medicine Veklury® from polymer capsules was reported. In this study, the effect of the physicochemical properties of chitosan on the size and stability of the produced structures is investigated, on the loaded amount of drug and on the kinetics of drug release in conditions close to the physiological ones. Microbiological studies of the capsules and their constituents were performed via in vitro assays against HCT-8 cell lines and human coronavirus HCoV-OC43. A detailed analysis was performed on the influence of the properties of produced capsules on cytotoxicity against the chosen cell line, as well as their effect on the replication cycle of the virus, the virucidal activity of the samples against the viability of the extracellular virions, and their effect on viral adsorption on the cell membrane.

Acquired AKT-inhibitor Resistance Is Mediated by ATP-binding Cassette Transporters in Endometrial Carcinoma.

Endometrial cancer is increasing in prevalence worldwide. It is treated with chemotherapy and radiotherapy, in addition to surgery, but the presence of treatment-resistant tumor cells remains a barrier to effective tumor control. The purpose of this study was to develop drug-resistant cell lines using triciribine, an AKT inhibitor, and investigate the mechanism of acquired resistance. Triciribine sensitivity assays were performed using Cell Counting Kit-8 (CCK-8) on eight endometrial cancer cell lines. The chosen cell lines were highly sensitive to chemotherapy and radiotherapy. A new triciribine-resistant cell line was established and found to be highly resistant to chemotherapy. Properties of the resistant cell line were identified using molecular and cell biological techniques including CCK-8 and quantitative PCR analysis. HEC-151 had the highest triciribine sensitivity (IC50 value of 0.7±0.1 μM) of the endometrial cancer cell lines tested. We established a triciribine-resistant cell line from HEC-151 by growing cells in the presence of increasing concentrations of triciribine up to 66.6 μM. The resistant HEC-151 cells changed to spindle-shaped morphology and importantly reduced triciribine sensitivity compared to the parental cell line. ABC transporters involved in drug efflux had significantly higher expression levels in ABCB1 (1.4±0.10 times higher), ABCC1 (11.4±0.22 times higher), and ABCC4 (4.5±0.42 times higher). In this study, we established a triciribine-resistant cell line from HEC-151 cells. Our data suggest that the mechanism of drug resistance in endometrial cancer cells is attributed to the increased expression of ABC transporters.

Clonogenic Survival RBE Calculations in Carbon Ion Therapy: The Importance of the Absolute Values of α and β in the Photon Dose-Response Curve and a Strategy to Mitigate Their Anticorrelation

The computation of the relative biological effectiveness (RBE) is a fundamental step in the planning of cancer radiotherapy treatments with accelerated ions. Numerical parameters derived analyzing the dose response of the chosen cell line after irradiation to photons (i.e., α and β, namely the linear and quadratic terms of the linear-quadratic model of cell survival) are generally used as input to biophysical models to predict the ion RBE. The α/β ratio for the photon exposure is generally regarded as an indicator of cell radiosensitivity. However, previous studies suggest that α/β might not be a sufficient parameter to model the RBE of relatively high linear energy transfer (LET) radiation such as carbon ions. For a fixed α/β, the effect of the absolute values of α and β on the computed RBE is underexplored. Furthermore, since α and β are anticorrelated during the fit of the photon-exposed in vitro survival data, different linear-quadratic fits could produce different sets of α and β, thus affecting the RBE calculations. This article reports the combined effect of the α/β ratio and the absolute values α and β on the RBE computed with the Mayo Clinic Florida microdosimetric kinetic model (MCF MKM) for 12C ions of different LET. Furthermore, we introduce a theory-based strategy to potentially mitigate the anticorrelation between α and β during the fit of the photon dose-response biological data.

The Expression and Activity of Rhodanese, 3-Mercaptopyruvate Sulfurtransferase, Cystathionine γ-Lyase in the Most Frequently Chosen Cellular Research Models.

This paper provides information concerning the activity and expression levels of three sulfurtransferases (STRs): rhodanese (TST, EC: 2.8.1.1), 3-mercaptopyruvate sulfurtransferase (MPST, EC: 2.8.1.2) and cystathionine γ-lyase (CTH, EC: 4.4.1.1) in various cell lines. Since very limited data are available in the scientific literature on this subject, the available data are included in this paper. These shortages often force the researchers to carry out their own screening tests that allow them to choose an appropriate model for their further studies. This work supplements the existing deficiencies in this area and presents the activity and expression of STRs in the eight most frequently chosen cell lines: the mouse mammary gland cell line (NMuNG, ATCC: CRL-1636), mouse mammary gland tumor (4T1, ATCC: CRL-2539), mouse fibroblast (MEF, ATCC: SCRC-1008), mouse melanoma (B16-F1, ATCC: CRL-6323), human colorectal adenocarcinoma (Caco-2, ATCC: HTB-37), human embryonic kidney (HEK-293, ATCC: CRL-1573), human osteosarcoma (MG-63, ATCC: CRL-1427) and rat myocardium (H9c2, ATCC: CRL-1446). Changes in STRs activity are directly related to the bioavailability of cysteine and the sulfane sulfur level, and thus the present authors also measured these parameters, as well as the level of glutathione (its reduced (GSH) and oxidized (GSSG) form) and the [GSH]/[GSSG] ratio that determines the antioxidant capacity of the cells. STRs demonstrate diverse functionality and clinical relevance; therefore, we also performed an analysis of genetic variation of STRs genes that revealed a large number of polymorphisms. Although STRs still provide challenges in several fields, responding to them could not only improve the understanding of various diseases, but may also provide a way to treat them.

Chemical Composition and Cytotoxic Activity of Eucalyptus torquata Luehm. and Eucalyptus salmonophloia F. Muell. Trunk Bark Essential Oils against Human SW620 and MDA-MB-231 Cancer Cell Lines.

In recent years, there has been a growing interest in the screening of natural active ingredients from Eucalyptus essential oils because of their evident importance in practical utility and their undeniable therapeutic properties. Based on this, the aim of the present study was to investigate the chemical profile of the essential oils of the trunk bark of Eucalyptus torquata Luehm. (ETEO), and E. salmonophloia F. Muell. (ESEO), growing in Tunisia. The in vitro cytotoxic properties of the extracted EOs were also evaluated against two human cancer cell lines: breast carcinoma cell lines MDA-MB-231 and colorectal cancer cell lines SW620. The analysis by gas chromatography coupled with mass spectrometry (GC/MS) led to the identification of 32 compounds from the ETEO, with the dominant constituents being the monoterpenes trans-myrtanol (73.4 %) and myrtenol (4.7 %), and the apocarotene (E)-β-ionone (3.9 %). In the case of ESEO, 29 compounds were identified with trans-myrtanol (25.0 %), decanoic acid (22.1 %), nonanoic acid (9.8 %), γ-elemene (6.5 %), γ-maaliene (5.5 %), and α-terpineol (5.3 %) as the main components. The cytotoxicity of EOs against the two chosen cell lines was tested using Crystal Violet Staining (CVS) assay and 5-fluorouracil as a reference drug. The two EOs exhibited a significant dose-dependent inhibition against the viability of the used cell lines. Their inhibitory effects were particularly observed towards SW620 colon carcinoma cells with IC50 values of 26.71±1.22 and 22.21±0.85 μg/mL, respectively, indicating that both oils were more cytotoxic for SW620 cells compared to MDA-MB-231 one.

Biaryl Sulfonamides Based on the 2-Azabicycloalkane Skeleton-Synthesis and Antiproliferative Activity.

In a search for new, selective antitumor agents, we prepared a series of sulfonamides built on bicyclic scaffolds of 2-azabicyclo(2.2.1)heptane and 2-azabicyclo(3.2.1)octane. To this end, aza-Diels–Alder cycloadducts were converted into amines bearing 2-azanorbornane or a bridged azepane skeleton; their treatment with sulfonyl chlorides containing biaryl moieties led to the title compounds. The study of antiproliferative activity of the new agents showed that some of them inhibited the growth of chosen cell lines with the IC50 values comparable with cisplatin, and some derivatives were found considerably less toxic for nonmalignant cells.

LASP2 functions as a potential prognostic factor and therapeutic target in nasopharyngeal carcinoma.

The purpose of this study was to investigate the potential effects of LIM and Src homology 3 (SH3) protein 2 (LASP2) on nasopharyngeal carcinoma (NPC) and the relevant mechanism. The expression of LASP2 in NPC patients and non-cancer patients in the control group was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The patients were divided into LASP2 high-expression group (n=30) and low-expression group (n=30), according to the median expression level of LASP2. Then, the expression of LASP2 was detected in the chosen cell lines by qRT-PCR. In qRT-PCR experiment, LASP2 was found up-expressed in NPC clinical samples and cell lines. Besides, LASP2 expression was associated with the clinical stage and distant metastasis of NPC. Next, the expression of LASP2 was downregulated by transfection of si-LASP using LipofectamineTM 3000 in 6-10B cells in vitro. The transfection effects of si-LASP2 were confirmed by qRT-PCR and Western-blot (WB) experiments. In supplementary experiments, decreased expression of LASP2 in cells could inhibit the cell biological functions, including invasion, migration, and epithelial-mesenchymal transition (EMT). This research discovers the promotion effect of LASP2 on NPC, suggesting that LASP2 could be used as a potential therapeutic target for NPC.

Abstract 3684: Basal Notch1 and Notch4 activation as potential markers of aggressiveness and sensitivity to Notch inhibition in human cancer cell lines

Abstract Background. Notch pathway is involved in tumor biology including cell proliferation, migration, drug resistance, and epithelial-to-mesenchymal transition (EMT). Notch activation leads to a proteolytic cleavage, followed by the release of the Notch intracellular domain (NICD), which translocates into the nucleus and activates target genes. Notch1 and Notch4 expressions were described as mutually exclusive in some tumors, with an increased aggressiveness linked to low Notch1/high Notch4 expression phenotype. What about the role of their basal activation in cancers? The aim of this work is to characterize the role of basal Notch1 and Notch4 pathway on cell proliferation, motility and sensitivity to Notch inhibition in human pancreatic (PDAC), head&neck (H&N), colorectal (CRC), cholangio (CK), and hepato (HCC) carcinoma cell lines. Materials and Methods. Cell lines were selected regarding their basal expression of NICD4 using western blot (one low/one high Notch4 basal activation) for each tumor type. Chosen cell lines were then characterized, by western blot and/or RT-qPCR, for Notch signaling (NICD1, HES1, NUMB), EMT status (vimentin, E-cadherin) and survival pathways (pAKT, pERK). Basal cell proliferation and motility were studied using MTT and wound-healing assay. The effects of a Notch inhibitor, PF-03084014, were assessed on cell proliferation and Notch signaling pathway by MTT and western blot. Results. In PDAC, H&N, and CRC cell lines, high NICD4 expression was associated with low NICD1, HES1, and NUMB expressions, whereas the opposite was observed in low NICD4 expressing cells. Interestingly, all high NICD4/low NICD1 cells displayed a mesenchymal phenotype with high vimentin and low E-cadherin expressions. This phenotype was also associated with an increased proliferation rate and basal cell motility in all tumor types, except for HCC. In addition, basal ERK phosphorylation was increased in the aggressive cells (high NICD4/low NICD1). In PDAC, H&N, and CRC cell lines, PF-03084014 showed higher antiproliferative effects in high NICD4/low NICD1 cells compared to low NICD4/high NICD1 cells. In all cell lines, PF-03084014 displayed no effect on Notch4 activation, but abrogated Notch1 activation in all low NICD4/high NICD1 cells. Interestingly, PF-03084014 decreased HES1 expression in all cells, with increased effects in the most sensitive cells (high NICD4/low NICD1). Further analysis of PF-04084014 effects on cell signaling pathways will be displayed at the conference. Conclusions. In this study, high Notch4 activation is shown to be correlated with low NICD1, HES1 and NUMB expressions. This phenotype is associated with an increased aggressiveness of human cancer cell lines, and an increased sensitivity to Notch inhibition. As expected, the strongest HES1 inhibition was observed in cell lines with an increased sensitivity to Notch inhibition. Since the inhibition of Notch pathway is an interesting topic for anticancer therapy, this study could help to select tumor types that may be good candidate for Notch inhibitors in the clinics. Citation Format: Lucile Astorgues-Xerri, Matthieu Martinet, Eric Raymond, Annemilaï Tijeras-Raballand. Basal Notch1 and Notch4 activation as potential markers of aggressiveness and sensitivity to Notch inhibition in human cancer cell lines [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3684.

Abstract B27: A DARPin-based toolbox to understand and treat RAS-addicted cancers

Abstract The repeating failure of small molecules as specific inhibitors of KRAS has drawn the attention to macromolecular structures, which can recognize their target with high affinity and specificity. Designed Ankyrin Repeat Proteins (DARPins) are multipurpose alternative affinity reagents that have proved to recognize targets with exceptional specificities and selectivity that often surpass those of antibodies. Due to their additional outstanding ability to act intracellularly within living cells and their recognition of structural rather than linear epitopes, DARPins have enabled a multitude of more advanced projects. In-house selections against GTP- or GDP-loaded KRAS were performed, and several hundred different DARPins could be identified. In subsequent validations, we not only analyzed their affinities, but also focused on the essential features of cross-reactivities, the recognition of various epitopes and their biologic functionalities. A number of candidates directly interfere with the RAS-Raf interaction and SOS-mediated nucleotide exchange by binding to the identical epitope on RAS as proved by crystal structures. One of these lead candidates with an affinity of 10 nM was used to elucidate whether anti-KRAS DARPins can mediate a biologic effect in model systems of human cancers. For this purpose, we chose cell lines categorized as KRAS dependent to generate stable cell lines expressing a highly active anti-KRAS DARPin under an inducible promotor, and it revealed potent antitumor activity, reducing the proliferation, colony formation and anchorage-independent growth. We could furthermore show that the observed effect resulted from reduced signaling of KRAS through its downstream MEK-ERK and PI3K-AKT pathways and the induction of apoptosis. Importantly, this anti-KRAS DARPin was shown to have no effects in the immortalized cell line HEK293T. In addition to traditional knockdown approaches, this model could be used to assess RAS dependency of human cancers. Combined with various suitable intracellular delivery techniques currently under development in our laboratory, the potential of this model will be further investigated for its potential in the treatment of solid tumors in a mouse model. In our broad effort many additional DARPins were identified that recognize different, nonoverlapping epitopes and most likely will interfere with other essential functions, such as the nanoclustering of RAS. This toolbox also allows the development of various biosensors in living cells. This large set of binders has the potential not just to help the community to gain detailed insights into the various functions and of RAS, but also might highlight novel vulnerabilities and innovative ways to finally make these key players druggable. Citation Format: Jonas N. Kapp, Jonas V. Schaefer, Wouter Verdurmen, Gabriela Nagy, Ralph Degen, Patrick Ernst, Claudia Scholl, Andreas Plückthun. A DARPin-based toolbox to understand and treat RAS-addicted cancers [abstract]. In: Proceedings of the AACR Special Conference on Targeting RAS-Driven Cancers; 2018 Dec 9-12; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(5_Suppl):Abstract nr B27.

Neutron Autoradiography Combined With UV-C Sensitization: Toward the Intracellular Localization of Boron.

Our group has reported the imprint formation of biological material on polycarbonate nuclear track detectors by UV-C exposure, which is used as an approach to simultaneously visualize cell imprints and nuclear tracks coming from the boron neutron capture reaction. Considering that the cell nucleus has a higher UV-C absorption than the cytoplasm and that hematoxylin preferentially stains the nucleus, we proposed to enhance the contrast between these two main cell structures by hematoxylin staining before UV-C sensitization. In this study, several experiments were performed in order to optimize UV-C exposure parameters and chemical etching conditions for cell imprint formation using the SK-BR-3 breast cancer cell line. The proposed method improves significantly the resolution of the cell imprints. It allows clear differentiation of the nucleus from the rest of the cell, together with nuclear tracks pits. Moreover, it reduces considerably the UV-C exposure time, an important experimental issue. The proposed methodology can be applied to study the boron distribution independently from the chosen cell line and/or boron compounds.

Long noncoding RNA TPTE2P1 promotes the migration and invasion of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a common malignant tumor that poses a serious threat to human health and life. Metastasis is one of the reasons for high rate of relapse. Due to the lack of effective treatment, the prognosis of HCC patients is far from satisfactory. The aim of this study was to investigate the role of long non-coding RNA (lncRNA)-TPTE2P1 in HCC development. Moreover, we aimed to search for new biomarkers which could predict the metastasis and provide novel therapeutic strategies for HCC. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to evaluate the expression level of lncRNA-TPTE2P1 in both HCC tissues and cell lines. Wound-healing assay and transwell assay were applied to determine the ability of cell migration. Meanwhile, transwell matrigel assay was applied to detect the invasion of HCC cells. The protein expressions of E-cadherin, Vimentin and N-cadherin in chosen cell lines were detected by Western blotting. Results found that lncRNA-TPTE2P1 was overexpressed in both HCC tissues and cell lines. Further analysis revealed that overexpression of lncRNA-TPTE2P1 was correlated with tumor size, distant metastasis, differentiation degree, as well as tumor node metastasis (TNM) stage of HCC patients. Subsequent wound-healing assay, transwell assay and Matrigel assay confirmed that down-regulated lncRNA-TPTE2P1 could significantly suppress the invasion and migration of cells. However, up-regulation of lncRNA-TPTE2P1 showed the opposite results. Moreover, lowly expressed lncRNA-TPTE2P1 significantly decreased the protein levels of E-cadherin, Vimentin and N-cadherin. These results indicated that lncRNA-TPTE2P1 might stimulate the migration and invasion of HCC cells by promoting epithelial-mesenchymal transition (EMT). In summary, our results suggested that lncRNA-TPTE2P1 functioned as an oncogene in HCC. Meanwhile, lncRNA-TPTE2P1 stimulated HCC cell migration and invasion by promoting EMT. LncRNA-TPTE2P1 might play a vital role in the development and progression of HCC. Our findings demonstrated that lncRNA-TPTE2P1 could serve as an early biomarker of metastasis and therapeutic target for HCC.

Synthesis, characterization, cytotoxic activity, and 19F NMR spectroscopic investigations of (OC-6-33)-diacetato(ethane-1,2-diamine)bis(3,3,3-trifluoropropanoato)platinum(IV) and its platinum(II) counterpart

Tetracarboxylatoplatinum(IV) complexes are interesting representatives of potential anticancer active platinum(IV) drugs. Revealing higher kinetic inertness in comparison to their cytotoxic platinum(II) counterparts, they offer an opportunity to reduce toxic and deactivating side reactions. Platinum(IV) complexes are generally considered as prodrugs, which have to be reduced in order to exert their anticancer activity. Two model complexes, a platinum(II) and platinum(IV) complex featuring two equatorial 3,3,3-trifluoropropanoato (tfpa) and in the case of the latter two axial acetato ligands, were synthesized and characterized in detail by multinuclear (1H, 13C, 15N, 19F, 195Pt) one- and two-dimensional NMR spectroscopy, high-resolution electrospray ionization mass spectrometry, elemental analysis and X-ray diffraction analysis. Cytotoxicity was evaluated in three human cancer cell lines (A549, SW480 and CH1/PA-1) by the MTT colorimetric assay, exhibiting IC50 values down to the low micromolar range. The fluorinated ligands in equatorial positions allowed detailed and highly sensitive 19F NMR spectroscopic investigations. Besides reaction studies in the presence of small molecules, e.g. ascorbic acid and 5′-GMP, the reduction of the platinum(IV) complex was additionally investigated in two cancer cell extracts deriving from the cell lines SW480 and CH1/PA-1 in order to estimate the intracellular reduction behavior. A significant increase in the rate of reduction in cell extracts (depending on the chosen cell line) compared to the reduction with ascorbic acid could be demonstrated.

Widespread Expression of Hedgehog Pathway Components in a Large Panel of Human Tumor Cells and Inhibition of Tumor Growth by GANT61: Implications for Cancer Therapy.

The sonic Hedgehog/GLI signaling pathway (HH) is critical for maintaining tissue polarity in development and contributes to tumor stemness. Transcription factors GLI1–3 are the downstream effectors of HH and activate oncogenic targets. To explore the completeness of the expression of HH components in tumor cells, we performed a screen for all HH proteins in a wide spectrum of 56 tumor cell lines of various origin using Western blot analysis. Generally, all HH proteins were expressed. Important factors GLI1 and GLI2 were always expressed, only exceptionally one of them was lowered, suggesting the functionality of HH in all tumors tested. We determined the effect of a GLI inhibitor GANT61 on proliferation in 16 chosen cell lines. More than half of tumor cells were sensitive to GANT61 to various extents. GANT61 killed the sensitive cells through apoptosis. The inhibition of reporter activity containing 12xGLI consensus sites by GANT61 and cyclopamine roughly correlated with cell proliferation influenced by GANT61. Our results recognize the sensitivity of tumor cell types to GANT61 in cell culture and support a critical role for GLI factors in tumor progression through restraining apoptosis. The use of GANT61 in combined targeted therapy of sensitive tumors, such as melanomas, seems to be immensely helpful.

Culture characters, genetic background, estrogen/progesterone receptor expression, and tumorigenic activities of frequently used sixteen endometrial cancer cell lines

BackgroundThis study aimed to determine the in vitro and in vivo properties of sixteen frequently used endometrial cancer (EC) cell lines, including the cell proliferation rate, morphology, hormone receptor expression patterns, PTEN, hMLH1 expression, p53 mutation, karyotype, and tumorigenicity in mouse xenograt model. MethodsTwelve type I (AN3, ECC-1, EN, EN-1, EN-11, HEC-1A, HEC1B, Ishikawa, KLE, MFE-280, MFE-296, MFE-319) and four type II (ARK1, ARK2, HEC-155/180, SPEC-2) endometrial cancer cell lines were studied. Cell proliferation and morphology were determined using cell growth curves and light microscopy, respectively. Real-time PCR was performed to measure the mRNA levels of target genes. Denaturing High Performance Chromatography (DHPLC) screening and PCR/sequencing were performed to identify p53 mutations. G-banding was applied for karyotyping. Tumorigenicity was evaluated using mouse xenograft. ResultsThe population doubling time of the cell lines ranged between 19 and 41 h. Ishikawa, ECC-1, and MFE-280 have high while AN3 and EN1 have low expression of ER-α and ER-β. Expression of total PR and PR-B uniformly decreased in all type II cell lines and several type I cell lines (AN3, HEC-1A, HEC1B, KLE, EN-1). Regression analyses revealed significant correlations between PR-B and total PR (p < .001), between isoforms ER-α and ER-β (p < .001), and between total PR and ER (p < .001), mRNA levels in type I cell lines. p53 mutations were detected in exons 5–8 of seven out of twelve type I and one out of four type II cell lines. PTEN expression was more uniformly suppressed in type II than type I cells, while hMLH1 did not show this pattern. All the five cell lines tested contained severe karyotype abnormalities. Mouse xenograft results indicated that HEC-1A, HEC-1B and EN-1 type I as well as ARK1 and ARK2 type II cell lines had potent tumorigenic activities. Low PR-B and ER-α expression in type I cell lines were associated with high tumorigenic activity. ConclusionsThis study provides resource information on EC cell lines commonly used in laboratories, which could be used for choosing cell lines suitable for specific research purposes. The results of karyotype analysis and p53 mutation together with hormone receptor expression pattern and morphology comparison strongly suggested an independent nature of these cell lines, excluding the possibility of cross-contamination between cell lines.Additionally, this information suggests potential directions for future studies on the pathogenic mechanisms of endometrial cancer.

Matrix metalloproteinase (MMP) and immunosuppressive biomarker profiles of seven head and neck squamous cell carcinoma (HNSCC) cell lines.

Biomarkers like programmed death ligand-1 (PDL1) have become a focal point for immunotherapeutic checkpoint inhibition in head and neck squamous cell carcinoma (HNSCC). However, it's only part of the total immunosuppressive biomarker profile of HNSCC cells. Matrix metalloproteinases (MMPs) are enzymes that break down the basement membrane allowing cancer cells to metastasize and play an important role in the tumor microenvironment. MMPs can also activate certain cytokines, growth factors, and chemokines post-translationally. The objective of this study was to determine MMP and biomarker profiles of seven different HNSCC cell lines. Authenticated cell lines were grown in minimal media at 1×106 viable cells/mL and incubated at 37 °C. After 24 hrs supernatants were collected, and adhering cells were lysed. Multiplex immunoassays were used to determine MMP1, MMP7, MMP9, IL-6, VEGFA, IL-1α, TNF-α, GM-CSF, IL-1RA, and IL-8 concentrations in supernatants. ELISAs were used to determine PDL1, CD47, FASL, and IDO concentrations in cell lysates. A one-way ANOVA was fit to examine log-transformed concentrations of biomarkers between seven HNSCC cell lines, and pairwise group comparisons were conducted using post- hoc Tukey's honest significance test (α=0.05). Significant differences (P<0.05) in MMP and biomarker concentrations were found between the seven HNSCC cell lines. For example, MMP9 was highest in SCC25 and UM-SCC99, MMP7 was highest in SCC25 and UM-SCC19, and MMP1 was highest in SCC25. These results suggest different patients' HNSCC cells can express distinct profiles of select biomarkers and MMPs, which could be due to metastatic stage of the cancer, primary tumor site, type of tissue the tumor originated from, or genomic differences between patients. MMP and biomarker expression profiles should be considered when choosing cell lines for future studies. The results support the reason for personalized medicine and the need to further investigate how it can be used to treat HNSCC.

Metal(II) complexes of bioactive aminonaphthoquinone-based ligand: synthesis, characterization and BSA binding, DNA binding/cleavage, and cytotoxicity studies

An aminonaphthoquinone ligand, L, and its metal complexes of general formula [MLCl2] {M = Co(II), Ni(II), Cu(II) and Zn(II)} have been synthesized and characterized by analytical and spectral techniques. Tetrahedral geometry has been assigned to Ni(II) and Zn(II) complexes and square planar geometry to Co(II) and Cu(II) complexes on the basis of electronic spectral and magnetic susceptibility data. The binding of complexes with bovine serum albumin (BSA) is relatively stronger than that of free ligand and alters the conformation of the protein molecule. Interaction of these complexes with CT-DNA has been investigated using UV-Vis and fluorescence quenching experiments, which show that the complexes bind strongly to DNA through intercalative mode of binding (Kapp 105 M−1). Molecular docking studies reiterate the mode of binding of these compounds with DNA, proposed by spectral studies. The ligand and its complexes cleave plasmid DNA pUC18 to nicked (Form II) and linear (Form III) forms in the presence of H2O2 oxidant. The in vitro cytotoxicity screening shows that Cu(II) complex is more potent against MCF-7 cells and Zn(II) complex exhibits marked cytotoxicity against A-549 cells equal to that of cisplatin. Cell imaging studies suggested apoptosis mode of cell death in these two chosen cell lines.

Found in Translation: Maximizing the Clinical Relevance of Nonclinical Oncology Studies.

Purpose: The translation of nonclinical oncology studies is a subject of continuous debate. We propose that translational oncology studies need to optimize both pharmacokinetic (drug exposure) and pharmacodynamic (xenograft model) aspects. While improvements in pharmacodynamic translatability can be obtained by choosing cell lines or patient-derived xenograft models closer to the clinical indication, significant ambiguity and variability exists when optimizing the pharmacokinetic translation of small molecule and biotherapeutic agents.Experimental Design and Results: In this work, we propose a pharmacokinetic-based strategy to select nonclinical doses for approved drug molecules. We define a clinically relevant dose (CRD) as the dosing regimen in mice that most closely approximates the relevant pharmacokinetic metric in humans. Such metrics include area under the time-concentration curve and maximal or minimal concentrations within the dosing interval. The methodology is applied to six drugs, including targeted agents and chemotherapeutics, small and large molecules (erlotinib, dasatinib, vismodegib, trastuzumab, irinotecan, and capecitabine). The resulting efficacy response at the CRD is compared with clinical responses.Conclusions: We conclude that nonclinical studies designed with the appropriate CRDs of approved drug molecules will maximize the translatability of efficacy results, which is critical when testing approved and investigational agents in combination. Clin Cancer Res; 23(4); 1080-90. ©2016 AACR.

Branching within branching: A model for host–parasite co-evolution

We consider a discrete-time host–parasite model for a population of cells which are colonized by proliferating parasites. The cell population grows like an ordinary Galton–Watson process, but in reflection of real biological settings the multiplication mechanisms of cells and parasites are allowed to obey some dependence structure. More precisely, the number of offspring produced by a mother cell determines the reproduction law of a parasite living in this cell and also the way the parasite offspring is shared into the daughter cells. In this article, we provide a formal introduction of this branching-within-branching model and then focus on the property of parasite extinction. We establish equivalent conditions for almost sure extinction of parasites and find a strong relation of this event to the behavior of parasite multiplication along a randomly chosen cell line through the cell tree, which forms a branching process in random environment. We then focus on asymptotic results for relevant processes in the case when parasites survive. In particular, limit theorems for the processes of contaminated cells and of parasites are established by using martingale theory and the technique of size-biasing. The results for both processes are of Kesten–Stigum type by including equivalent integrability conditions for the martingale limits to be positive with positive probability. The case when these conditions fail is also studied. For the process of contaminated cells, we show that a proper Heyde–Seneta norming exists such that the limit is nondegenerate.