Adenocarcinoma Tumor Cell Lines Research Articles

Integrative analysis indicates the potential values of ANKRD53 in stomach adenocarcinoma.

Ankyrin repeat domain 53 (ANKRD53) plays an important role in maintaining chromosome integrity and stability, and chromosome instability is associated with cancer. Through integrative analysis, this study investigates the potential value of ANKRD53 in stomach adenocarcinoma (STAD). RNA-seq and scRNA-seq data were used for integrative analysis based on online databases. Expression of ANKRD53 was confirmed by RT-PCR after bioinformatic analysis. Kaplan-Meier and Cox regression analyses were performed to evaluate the prognostic value of ANKRD53 in STAD. Gene set enrichment analysis (GSEA) was performed to evaluate ANKRD53-related signaling pathways. In addition, the interaction of ANKRD53 with immunity was also investigated. RT-PCR in STAD cell lines confirmed that ANKRD53 was downregulated in STAD samples compared to normal samples in the online databases. As an independent predictive biomarker, ANKRD53 was combined with other clinicopathological parameters to create a prognostic nomogram. Using GSEA, ANKRD53 was found to be involved in five pathways, including the TGF-β signaling pathway. Further investigation revealed that ANKRD53 was associated with immune checkpoint molecules, immunological pathways, and immunotherapy, in addition to MSI, TMB and neoantigens. In addition, scRNA-seq data revealed that ANKRD53 is mainly expressed in CD8+ T and dendritic cells. ANKRD53 is an important biomarker for STAD that deserves further attention.

MAGED4B is a Poor Prognostic Marker of Stomach Adenocarcinoma and a Potential Therapeutic Target for Stomach Adenocarcinoma Tumorigenesis.

Gastric cancer is the second most common cause of cancer death worldwide with poor overall prognosis. It is important to study the molecular mechanism of stomach adenocarcinoma (STAD). MAGED4B, a member of the melanoma antigen gene (MAGE) family, is highly expressed in many tumor cells and is associated with tumor progression. Its prognostic value in and the function of the encoded protein are still unclear. The data of 415 STAD tissues was retrieved from TCGA database, and the expression level of MAGED4B mRNA was evaluated. The correlation between the expression of MAGED4B mRNA and the progression free survival (PFS) time of STAD patients was evaluated by Kaplan Meier analysis. The STAD cell lines with overexpressed and silent MAGED4B were constructed, and the effects of MAGED4B on the viability, migration and proliferation were evaluated by the CCK-8, scratch test and EDU test. The flow cytometry was used to detect apoptosis with overexpressed and silent MAGED4B under the cisplatin treatment, and WB was used to detect the expressions of related proteins, such as TNF-α. The expression level of MAGED4B mRNA in the STAD tissues was higher than that in the normal tissues, and its high expression was related to poor PFS. The overexpression of MAGED4B in the STAD cell lines can promote the vitality, motility and proliferation of the STAD cells, while the silencing of MAGED4B can inhibit the above three cell functions of the STAD cells. The overexpression of MAGED4B can reduce the cisplatin induced apoptosis and increase the cisplatin IC50; the silencing of MAGED4B can promote the cisplatin induced apoptosis and reduce the cisplatin IC50. The overexpression of MAGED4B reduced the protein levels of TRIM27 and TNF- α. MAGED4B could be a valuable prognostic biomarker and a therapeutic target for gastric adenocarcinoma of great interest.

Alginate-Based Hydrogel as Delivery System for Therapeutic Bacterial RNase.

To deliver therapeutic proteins into a living body, it is important to maintain their target activity in the gastrointestinal tract after oral administration. Secreted ribonuclease from Bacillus pumilus (binase) has antitumor and antiviral activity, which makes it a promising therapeutic agent. This globular protein of small molecular weight (12.2 kDa) is considered as a potential agent that induces apoptosis of tumor cells expressing certain oncogenes, including colorectal and duodenum cancer. The most important problem of its usage is the preservation of its structure and target activity, which could be lost during oral administration. Here, we developed alginate microspheres reinforced with divalent cations and analyzed the enzyme release from them. Using methods of scanning electron microscopy, measurements of fluorescence, enzyme catalytic activity, and determination of viability of the duodenum adenocarcinoma tumor cell line, we characterized obtained microspheres and chose calcium as a biogenic ion-strengthening microsphere structure. Among such modified additivities as beta-casein, gelatin, and carbon nanotubes introduced into microspheres, only gelatin showed a pronounced increase in their stability and provided data on the prolonged action of enzyme release from microspheres into tumor cell culture medium during 48 h in an amount of about 70% of the loaded quantity.

Long noncoding RNA TUG1 upregulates VEGFA to enhance malignant behaviors in stomach adenocarcinoma by sponging miR-29c-3p.

BackgroundLong noncoding RNA (lncRNA) TUG1 has been reported to display a pivotal role in the tumorigenesis and malignant progression of various types of cancers, including stomach adenocarcinoma (STAD). However, the contribution of aberrant expression of TUG1 and the mechanism by which it serves as a competing endogenous RNA (ceRNA) in STAD remains largely obscure.MethodsThe human STAD cell lines (MGC‐803 and AGS), human normal gastric epithelial cell line (GES‐1), human umbilical vein endothelial cells (HUVECs), and human embryonic kidney cells (HEK293T) were purchased and cultured to investigate the roles of TUG1 in STAD. Twenty BALB/c nude mice were purchased to establish a xenograft model to explore the roles of TUG1 in vivo.ResultsBioinformatics analysis revealed that TUG1 was upregulated in STAD, of which expression was negatively and positively correlated with miR‐29c‐3p and VEGFA, respectively. Functional analyses indicated that TUG1 functioned as an oncogene to promote malignant behaviors (proliferation, migration, and angiogenesis) of STAD cells; whereas miR‐29c‐3p exerted the opposite role. Mechanistically, the interaction between miR‐29c‐3p with TUG1 and VEGFA was demonstrated. It was observed that miR‐29c‐3p could reverse the TUG1‐induced promotion effect on cell proliferation, migration, and angiogenesis in STAD. Furthermore, TUG1 overexpression promoted STAD cell proliferation, metastasis, and angiogenesis, whereas VEGFA silence restored these effects, both in vitro and in vivo.ConclusionThis finding confirmed that lncRNA TUG1 acts as a ceRNA for miR‐29c‐3p to promote tumor progression and angiogenesis by upregulating VEGFA, indicating TUG1 as a therapeutic target in STAD management.

Comprehensive Characterization of Integrin Subunit Genes in Human Cancers.

Although integrin subunit genes (ITGs) have been reported to be associated with some human cancer types, a systematic assessment of ITGs across human cancers is lacking. Hence, we performed comprehensive analyses to investigate mRNA expression, copy number variation (CNV), DNA methylation, mutation, and clinical landscapes of ITGs in more than 8000 cancer patients from The Cancer Genome Atlas (TCGA) dataset. Landscapes of ITGs were established across 20 human cancer types. We observed that ITGs are extensively dysregulated with heterogeneity in different system cancer types, part of which are driven by CNV, DNA hypomethylation or mutation. Furthermore, dysregulated prognosis-related ITGs were systematically identified in each cancer type, including ITGA11 in stomach adenocarcinoma (STAD). The models based on dysregulated ITGs with clinical relevance and TNM staging indexes are good indicators in STAD and head and neck squamous cell carcinoma. Finally, ITGA11 is overexpressed and associated with poor survival in STAD cases from the TCGA and additionally Gene Expression Omnibus cohorts. Functionally, ITGA11 knockdown inhibits malignant phenotypes in STAD cell lines AGS and MKN45, demonstrating the oncogenic role of ITGA11 in STAD. Together, this study highlights the important roles of ITGs in tumorigenesis as potential prognostic biomarkers, and provide an effective resource that identifies cancer-related genes of ITGs in human cancers.

High Endothelin Receptor Type A Expression as an Independent Prognostic Biomarker and Correlated with Immune Infiltrates in Stomach Adenocarcinoma.

BackgroundStomach adenocarcinoma (STAD) is the most common gastrointestinal cancer and is associated with high mortality worldwide. Endothelin receptor type A (EDNRA) is associated with guanine-nucleotide-binding (G) proteins and plays important roles in cellular processes and various diseases.PurposeTo investigate the prognosis value of EDNRA expression and its correlation with immune infiltrates in patients with STAD.MethodsThe association between clinical characteristics and EDNRA expression in STAD was analyzed using the Wilcoxon signed-rank test and logistic regression. The Kaplan–Meier plotter analysis and Cox regression were constructed to evaluate the influence of EDNRA on prognosis, and a receiver operating characteristic (ROC) curve and nomogram were constructed. Gene set enrichment analysis (GSEA) and single-sample gene set enrichment analysis (ssGSEA) were conducted to analyze the correlation between EDNRA and immune infiltrates. In addition, Oncomine, TIMER databases and qRT-PCR of STAD cell lines were used to verify the EDNRA expression in STAD.ResultsOur results revealed that EDNRA expression was significantly higher in patients with STAD than normal gastric tissues, and the results have been confirmed by RT-qPCR. KM-plotter analysis revealed that patients with STAD had shorter OS, FP, and PPS (P<0.001). Multivariate Cox analysis further confirmed that high EDNRA expression was an independent risk factor for OS in patients with STAD. Moreover, other clinicopathologic features were related with worse prognosis in STAD, including age, lymph nodes metastases and primary outcome. More importantly, ROC analysis also confirmed the diagnostic value, and a prognostic nomogram involving age, T, M, N classification, pathologic stage, residual tumor and EDNRA was constructed. GSEA revealed that high EDNRA expression was correlated with immunoregulatory interactions between lymphoid and non lymphoid cells pathways, natural killer cell activation involved in immune response, interleukin 1 receptor binding and pathways in cancer, and ssGSEA showed that EDNRA is correlated with macrophages and NK cells.ConclusionCollectively, EDNRA can be an independent prognostic biomarker and correlated with immune infiltration in stomach adenocarcinoma.

ClC-3/SGK1 regulatory axis enhances the olaparib-induced antitumor effect in human stomach adenocarcinoma

Currently, only a few available targeted drugs are considered to be effective in stomach adenocarcinoma (STAD) treatment. The PARP inhibitor olaparib is a molecularly targeted drug that continues to be investigated in BRCA-mutated tumors. However, in tumors without BRCA gene mutations, particularly in STAD, the effect and molecular mechanism of olaparib are unclear, which largely restricts the use of olaparib in STAD treatment. In this study, the in vitro results showed that olaparib specifically inhibited cell growth and migration, exerting antitumor effect in STAD cell lines. In addition, a ClC-3/SGK1 regulatory axis was identified and validated in STAD cells. We then found that the down-regulation of ClC-3/SGK1 axis attenuated olaparib-induced cell growth and migration inhibition. On the contrary, the up-regulation of ClC-3/SGK1 axis enhanced olaparib-induced cell growth and migration inhibition, and the enhancement effect could be attenuated by SGK1 knockdown. Consistently, the whole-cell recorded chloride current activated by olaparib presented the same variation trend. Next, the clinical data showed that ClC-3 and SGK1 were highly expressed in human STAD tissues and positively correlated (r = 0.276, P = 0.009). Furthermore, high protein expression of both ClC-3 (P = 0.030) and SGK1 (P = 0.006) was associated with poor survival rate in STAD patients, and positive correlations between ClC-3/SGK1 and their downstream molecules in STAD tissues were demonstrated via the GEPIA datasets. Finally, our results suggested that olaparib inhibited the PI3K/AKT pathway in STAD cells, and up-regulation of ClC-3/SGK1 axis enhanced olaparib-induced PI3K/AKT pathway inhibition. The animal experiments indicated that olaparib also exerted antitumor effect in vivo. Altogether, our findings illustrate that olaparib exerts antitumor effect in human STAD, and ClC-3/SGK1 regulatory axis enhances the olaparib-induced antitumor effect. Up-regulation of the ClC-3/SGK1 axis may provide promising therapeutic potential for the clinical application of olaparib in STAD treatment.

Establishment and characterization of an in vitro 3D ovarian cancer model for drug screening assays.

The acquired drug chemoresistance represents the main challenge of the ovarian cancer treatment. In addition, the absence of an adequate in vitro model able to reproduce the native tumor environment can contribute to the poor success rate of pre-clinical studies of new compounds. Three-dimensional (3D) culture models have been recently used for drug screening purposes due to their ability to reproduce the main characteristics of in vivo solid tumors. Here we describe the establishment and characterization of 3D ovarian cancer spheroids using different adenocarcinoma tumor cell lines (SKOV-3 and OVCAR-3 cells) in two different 3D scaffold-free methods: forced-floating in ultra-low attachment (ULA) plates and hanging drop (HD). Spheroids were evaluated in both 3D cultures in order to establish the best condition to perform the drug response analysis with Paclitaxel, a common drug used to treat ovarian cancer. SKOV-3 and OVCAR-3 spheroids with the desired characteristics (roundness close to 1.0 and diameter in the 200-500 μm range) were obtained using both methods after addition of the methylcellulose (MC) in the culture medium (0.25% and 0.5%, w/v). We also observed the presence of microvilli on the surface of the spheroids, higher presence of apoptotic cells and higher drug resistance, when compared with 2D cultures. The 3D cultures obtained seem to provide more reliable results in terms of drug response than those provided by 2D monolayer culture. The forced floating method was considered more suitable and straightforward to generate ovarian cancer spheroids for drug screening/cytotoxicity assays.

The saffron effects on expression pattern of critical self-renewal genes in adenocarcinoma tumor cell line (AGS)

Abstracts Background The aim of the current study was to determine the effects of various concentrations of saffron distillate (Crocus sativus) on expression of some self-renewal genes including OCT4, KLF, SOX2, NANOG and Nucleostemin in AGS (gastric adenocarcinoma) cell line. Material and methods The AGS cells were cultured in RPMI1640 medium contained saffron (20, 40 and 100 μg/mL) for 24, 48 and 72 h. Inhibition of cell proliferation was assessed by MTT. The percentage of apoptotic cells was determined by flow cytometry analysis using Annexin V/PI apoptosis detection kit. Total cellular RNA extracted and subjected to and cDNA synthesis and then, the expression of the desired genes was extracted and subjected to in both the control and test groups, using real time- PCR (RT-PCR). Data were analyzed by student t and repeated measure tests. Differences were considered significant if (P Findings There was a significant difference in cell viability, when various concentrations of saffron were used for 24, 48 and 72 h of incubation and apoptosis increased in a time-dependent manner. Flow cytometry results exhibited an obviously significant augmentation in apoptotic AGS cells. Real time- PCR our RT-PCR based findings indicated that saffron distillate has reduced the expression of OCT4, KLF, SOX2, NANOG and Nucleostemin genes in compare to untreated control cells. Conclusion According to the results of this study it may be concluded that saffron distillate can be considered as an inhibitor of cell division and regardingly inhibit cancer progression.

FKBP10 functioned as a cancer-promoting factor mediates cell proliferation, invasion, and migration via regulating PI3K signaling pathway in stomach adenocarcinoma.

As documented, the expression, biological roles, and prognostic significance of FKBP10 in stomach adenocarcinoma (STAD) have not been investigated till now. This drives us to detect the biological roles and clinical significance of FKBP10 in STAD. The expression level of FKBP10 was measured based on the data obtained from the TCGA, ONCOMINE, and GEPIA databases, and STAD cell lines. Through in vitro experiments, cell behaviors were investigated to evaluate the effects of FKBP10 on STAD. Moreover, the PI3K-AKT signaling pathway was measured. Relying on the data of TCGA, ONCOMINE, and GEPIA databases, and cancer cell lines, FKBP10 was up-regulated in STAD when compared with normals. The patients with low expression of FKBP10 had higher survival rate than those with high FKBP10 expression. After knockdown of FKBP10 in AGS cells, cell vitality, colony formation ability, and the migratory and invasive potential were inhibited. Western blotting analysis exhibited that knockdown of FKBP10 significantly reduced the expression level of p-AKT, and p-PI3K, but it did not influence the total expression level of AKT, and PI3K. FKBP10 might serve as a crucial player in gastric cancer, and targeting FKBP10 might provide clinical utility in gastric cancer in future.

Abstract 5195: Efficacy of histone deacetylase inhibitors in a 3D cell culture model

Abstract Epigenetic dysregulation, changes in gene expression not due to changes in the DNA sequence, is the underlying cause of many diseases including cancer. Identification of enzymatic chromatin-complex modulators that can restore chromatin homeostasis is critical to developing better therapeutic agents for both, hematologic and solid tumors. 3D cell culture models have been developed to overcome the limitations of monolayer in vitro assays, to include the complexity of the tumor microenvironment known to modulate epigenetic factors, such as cell-cell interaction and allowing for formations of metabolic gradients, factors known to play a role in epigenetic drug efficacy and resistance. In this study, we evaluated the efficacy of class I and II Inhibitors of histone deacetylases, SAHA and Trichostatin A, in a 3D cell culture in vitro assay across several human tumor carcinoma cell lines. The cell lines were grown in 3D cell cultures, and efficacy of the drugs was evaluated at 72 hours and ten days. All the cell lines grown as spheroids proved to be highly sensitive to SAHA and Trichostation A. AGS, a stomach adenocarcinoma cell line, showed the highest sensitivity to SAHA with an IC50 of 270 nM. Trichostatin A, on the other hand, displayed higher sensitivity to the ovarian carcinoma cell line, SKOV3, with an IC50 of 190 nM. In all cases, maximum potency was seen at the 10-day incubations with IC50s below 1 µM for all the cell lines in the panel. Drug-drug combinations with the epigenetic drugs were also studied, the results are discussed. Citation Format: Macarena Irigoyen, Gonzalo Castillo. Efficacy of histone deacetylase inhibitors in a 3D cell culture model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5195.

Cytotoxic effect of crab shell extracts on different tumor cell lines

Crab, a kind of marine animal that has a unique status in evolution, has many bioactive substances with special functions. In the present study, we investigated the antitumor effects of four different extracts derived from crab shell (aqueous and methanolic extracts of burned and normal crab shell powder). Aqueous and methanolic extracts of burned crab shell powder were shown to be cytotoxic and inhibit the in vitro proliferation of human laryngeal carcinoma tumor cell lines (Hep-2) and human Rhabdomyosarcoma cell line (RD) and one murine mammary adenocarcinoma tumor cell line (AMN3), in dose and time dependant manner. These results suggest that the burned crab shell powder have an antitumor effect that needs more investigations.

Circulating exosomes potentiate tumor malignant properties in a mouse model of chronic sleep fragmentation.

BackgroundChronic sleep fragmentation (SF) increases cancer aggressiveness in mice. Exosomes exhibit pleiotropic biological functions, including immune regulatory functions, antigen presentation, intracellular communication and inter-cellular transfer of RNA and proteins. We hypothesized that SF-induced alterations in biosynthesis and cargo of plasma exosomes may affect tumor cell properties.ResultsSF-derived exosomes increased tumor cell proliferation (~13%), migration (~2.3-fold) and extravasation (~10%) when compared to exosomes from SC-exposed mice. Similarly, Pre exosomes from OSA patients significantly enhanced proliferation and migration of human adenocarcinoma cells compared to Post. SF-exosomal cargo revealed 3 discrete differentially expressed miRNAs, and exploration of potential mRNA targets in TC1 tumor cells uncovered 132 differentially expressed genes that encode for multiple cancer-related pathways.MethodsPlasma-derived exosomes from C57/B6 mice exposed to 6 wks of SF or sleep control (SC), and from adult human patients with obstructive sleep apnea (OSA) before (Pre) and after adherent treatment for 6 wks (Post) were co-cultured with mouse lung TC1 or human adenocarcinoma tumor cell lines, respectively. Proliferation, migration, invasion, endothelial barrier integrity and extravasation assays of tumor cells were performed. Plasma mouse exosomal miRNAs were profiled with arrays, and transcriptomic assessments of TC1 cells exposed to SF or SC exosomes were conducted to identify gene targets.ConclusionsChronic SF induces alterations in exosomal miRNA cargo that alter the biological properties of TC1 lung tumor cells to enhance their proliferative, migratory and extravasation properties, and similar findings occur in OSA patients, in whom SF is a constitutive component of their sleep disorder. Thus, exosomes could participate, at least in part, in the adverse cancer outcomes observed in OSA.

Purification of Cucurbitacins D, E, and I from Ecballium Elaterium (L.) A. Rich Fruits and Study of Their Cytotoxic Effects on the AGS Cell Line

Background:The plant Ecballium elaterium (L.) A. Rich, belongs to the Cucurbitaceae family which occupies an important position in traditional medicine prescriptions. It has been reported that a freeze-dried aqueous extract of E. elaterium fruits has cytotoxic effects on the AGS human stomach adenocarcinoma cell line. We here focused on anticancer effects of the main chemicals purified from E. elaterium fruits.Materials and Methods:We isolated cucurbitacins D, E, and I from chloroform, and ethyl acetate fractions of a methanolic extract of E. elaterium fruits and assessed their cytotoxic effects on the AGS cell line by MTT assay. The methanolic extract was fractionated to petroleum ether, chloroform, and ethyl acetate fractions. The compounds isolated by column chromatography were identified by NMR spectroscopy.Results:After 24 h of incubation with AGS cells, the IC50 values were 0.3, 0.1, and 0.5 μg/ml for cucurbitacins D, E, and I respectively.Conclusions:This finding suggests that because of its cucurbitacins, E. elaterium fruit may have some cytotoxic effects on gastric cancer cells. Also, compared with D and I, cucurbitacin E showed greater potency in this regard.

Spiculisporic acid analogues of the marine-derived fungus, Aspergillus candidus strain HDf2, and their antibacterial activity.

Two novel antibiotic spiculisporic acid analogues, named as spiculisporic acid F (1) and G (2), and two known compounds, (-)-spiculisporic acid (3) and secospiculisporic acid B (4), were isolated by bioactivity-guided fractionation from the fermentation broth of the sea urchin-derived Aspergillus candidus strain HDf2. Their structures were unambiguously established by comprehensive analysis of 1D and 2D NMR, and high-resolution MS spectra, and by comparison with known compounds. Biological experiments demonstrated that compounds 1 and 2 displayed antibacterial activity against Gram-negative Pseudomonas solanacearum and Gram-positive Staphylococcus aureus, but showed no cytotoxicity against SGC-7901 human gastric adenocarcinoma and SPC-A-1 human lung adenocarcinoma tumor cell lines. This is the first critical evidence identifying spiculisporic acid derivatives as a potential bio-control agent for the soil borne pathogen P. solanacearum (E. F. Smith) Smith. These findings provide further insight into the chemical and biological activity diversity of this class of compounds.

Efficient synthesis of panaxadiol derivatives using continuous-flow microreactor and evaluation of anti-tumor activity

Abstract An efficient method has been developed for the synthesis of a series of (20R)-panaxadiol derivatives (4a–w) using a continuous-flow microreactor. The antitumor activities of the newly synthesized compounds were evaluated in vitro in two human prostate adenocarcinoma tumor cell lines (i.e., PC-3 and LNCaP cells), and their cytotoxicities were evaluated using a standard MTT assay. Compounds 4c, 4h, 4p, 4q and 4s exhibited higher antitumor activities toward PC-3 cell line than panaxadiol, which was used as a reference standard.

Chemical Constituents of Ficus linearifolia and Ficus triangularis

Ficus linearifolia Elmer is an endemic Philippine fig tree distributed in the forests of Luzon, Negros, and Leyte at altitudes up to 900 m above sea level [1]. It was earlier considered as a mere variety of Ficus botryocarpa Miq. [2]. This fig tree was first described in 1907 as an erect tree that reaches a height of 20 m and bears linear lanceolate leaves that are more or less scattered but usually confined towards ends of the branchlets. It grows in low damp wooded ground or along streams in deep ravines at the base of Mt. Banahao (Lucban, Luzon Island) at 750 m above sea level [3]. This fig tree is locally known in the Philippines as daing-daing [4]. Ficus triangularis, commonly known as triangular fig and sweetheart tree, is a native to tropical Africa. It was recently introduced in the Philippines as an ornamental tree [5]. This study was conducted as part of our research on the chemical constituents of the genus Ficus. We earlier reported the isolation of a new neohopane [6] from Ficus pumila. We also reported the isolation of terpenoids and sterols from the endemic Philippine trees, Ficus pseudopalma and Ficus ulmifolia [7]. Recently, we reported the isolation of terpenoids and sterols from another endemic Philippine tree, Ficus odorata. -Sitosteryl-3 -glucopyranoside-6-O-palmitate from the leaves of F. odorata exhibited cytotoxicity against human stomach adenocarcinoma cell line (AGS) with 60.28% growth inhibition [8]. There are no reported chemical constituents and biological activities of F. linearifolia and F. triangularis. We report herein the isolation and structure elucidation of 1–9 from the dichloromethane extract of the air-dried leaves of Ficus linearifolia and 4–13 from the leaves of F. triangularis. To the best of our knowledge, this is the first report on the isolation of these compounds from these trees. Silica gel chromatography of the dichloromethane extract of the leaves of F. linearifolia afforded 1–9. The structure of 1 was elucidated by extensive 1D and 2D NMR spectroscopy and confirmed by comparison of their 13C NMR data with those of 11 ,12 -epoxyurs-14-en-3 -yl acetate [9]. The structures of 2–9 were identified by comparison of their 13C NMR data with those reported in the literature for -amyrin (2) [10], -amyrin (3) [10], squalene (4) [11], -sitosterol (5) [12], -stigmasterol (6) [12], polyprenol (7) [13], linoleic acid (8) [14], lutein (9) [15]. The leaves of Ficus linearifolia were collected from the foot of Mt. Banahaw, at around 600 m above sea level (masl), at Avila s Farm, Brgy Palola, Lucban, Quezon, Philippines in March 2012. Voucher specimens were authenticated by Danilo N. Tandang of Philippine National Museum. The air-dried leaves (310 g) of F. linearifolia were soaked in CH2Cl2 for 3 days and then filtered. The filtrate was concentrated under vacuum to afford the crude extract (6 g), which was chromatographed in increasing proportions of acetone in CH2Cl2 at 10% increments by volume as eluents. The CH2Cl2 fraction was rechromatographed (4 ) in petroleum ether to afford 4 (8 mg). The 10% acetone in CH2Cl2 fraction from the chromatography of the crude extract was rechromatographed (3 ) in 7.5% EtOAc in petroleum ether to afford 1 (12 mg). The 20% acetone in CH2Cl2 fraction from the chromatography of the crude extract was rechromatographed (4 ) in 10% EtOAc in petroleum ether to afford 7 (10 mg). The 30% acetone in CH2Cl2 fraction was rechromatographed (3 ) in 15% EtOAc in petroleum ether to afford 8 (15 mg) and a mixture of 2 and 3 (9 mg).

Chemical constituents of Ficus odorata

A dichloromethane extract of the leaves of Ficus odorata (Moreaceae family) contains l-sitosteryl3β-glucopyranoside-6'-O-palmitate (1), squalene (2), lutein (3), α-amyrin acetate (4), lupeol acetate (5), and β-carotene (6). The structure of 1 was elucidated by extensive 1D and 2D NMR spectroscopy. Compound 1 was tested for cytotoxicity using sulforhodamine B assay against the human lung adenocarcinoma epithelial cell line (A549), human stomach adenocarcinoma cell line (AGS), human colon adenocarcinoma (HT29) and human prostate cancer cell line (PC3). It exhibited cytotoxicity against AGS cell line with 60.28% growth inhibition

CoMFA and CoMSIA Studies of 1,2-dihydropyridine Derivatives as Anticancer Agents

Taking advantage of our in-house experimental data on 3-cyano-2-imino-1, 2-dihydropyridine and 3-cyano-2- oxo-1,2-dihydropyridine derivatives as inhibitors of the growth of the human HT-29 colon adenocarcinoma tumor cell line, we have established a highly significant CoMFA and CoMSIA models (q2cv=0.70/0.639). The models were investigated to assure their stability and predictivity (r2pred=0.65/0.61) and successfully applied to design two new potential cell growth inhibitory agents with IC50s in the submicromolar range.

Design, synthesis and biological evaluation of novel pyridine derivatives as anticancer agents and phosphodiesterase 3 inhibitors

Two series of 4,6-diaryl-2-imino-1,2-dihydropyridine-3-carbonitriles and their isosteric 4,6-diaryl-2-oxo-1,2-dihydropyridine-3-carbonitriles were synthesized through a combinatorial approach. The prepared analogues were evaluated for their in vitro capacity to inhibit PDE3A and the growth of the human HT-29 colon adenocarcinoma tumor cell line. Compound 6-(4-bromophenyl)-4-(2-ethoxyphenyl)-2-imino-1,2-dihydropyridine-3-carbonitrile (Id) exhibited the strongest PDE3 inhibition when cGMP but not cAMP is the substrate with a IC 50of 27 μM, which indicates a highly selective mechanism of enzyme inhibition. On the other hand, compound 6-(1,3-benzodioxol-5- yl)-4-(2-ethoxyphenyl)-2-imino-1,2-dihydropyridine-3-carbonitrile ( Ii) was the most active in inhibiting colon tumor cell growth with a IC 50 of 3 μM. The electronic effects, steric effects and conformational aspects of Id seem to be the most crucial for the PDE3 inhibition. Meanwhile, steric factors and the H-bonding capability seem to be the most important factors for tumor cell growth inhibitory activity. Conversely, there is no direct correlation between PDE3 inhibition and anticancer activity for the prepared compounds. An in silico docking experiment indicates the potential involvement of other potential molecular targets such as PIM-1 kinase to explain its tumor cell growth inhibitory activity.