Oil palm phenolics (OPP) or Palm Juice (PJ), a water soluble extract from the palm fruit (Elaies guineensis) has been documented to have anti-carcinogenic activities in various cancer types.
To ...investigate OPP effects in pancreatic cancer (PaCa) cells, two PaCa cell lines (PANC-1 and BxPC-3) were treated with different OPP doses. The anti-proliferative, apoptotic and anti-invasive properties of OPP were evaluated using MTS, cytoplasmic histone-DNA fragmentation and matrigel invasive assays, respectively.
OPP suppressed PaCa proliferation in a dose-dependent manner. Its anti-invasive effects were validated by decreased expressions of MMP-9 and VEGF. Cell-cycle analysis demonstrated that cells were arrested in the S phase. OPP-induced apoptosis was associated with decrease in survivin and Bcl-XL expressions and increased expression of cleaved caspase-3, caspase-9 and PARP.
Overall, our results demonstrate the anti-tumor effects of OPP on PaCa cells, providing initial evidence towards its potential therapeutic use.
Abstract
INTRODUCTION: In a recent study, we showed that SLC1A5, a key glutamine (Gln) transporter, regulates tumor growth and survival in NSCLC. Here we sought to characterize the expression of ...other Gln transporters in lung cancer and to test whether the inhibition of SLC1A5 activity induced a change in expression of other amino acid transporters in NSCLC.
METHODS: Neutral amino acid transporters reported in the cancer literature were evaluated in publically available lung cancer gene expression datasets. Among those, SLC38A1, SLC1A4, SLC1A5, SLC7A5, SLC7A11, SLC7A8 were selected for the in vitro validation. The expression of these genes was evaluated in two lung squamous carcinoma cell lines (HCC15, H226) and one adenocarcinoma cell line (A549) by RT-PCR and then quantified by qPCR, using SYBR® Green Supermix protocol. The expression of the same genes was assessed in response to transient SLC1A5 knockdown in HCC15 and A549 cell lines by qPCR.
RESULTS: Amino acid transporters SLC38A1, SLC1A5, SLC7A5 and SLC7A11 are expressed in NSCLCs, a finding that was confirmed in the three cell lines studied. SLC1A4 was found to have the lowest expression and SLC7A8 no expression in any of cell lines. In response to SLC1A5 transient knockdown in A549, SLC38A1, SLC1A4 and SLC7A5 were significantly up-regulated (p<0.02); in contrast, in HCC15, SLC38A1, SLC1A4 and SLC7A5 were significantly down-regulated (p<0.02) and SLC7A11 was up-regulated (p<0.13).
CONCLUSION: Our results show that SLC38A1, SLC1A5, SLC7A5 and SLC7A11 have the highest level of gene expression in NSCLC. We identified a possible compensatory mechanism to satisfy Gln transport and cellular function upon SLC1A5 silencing involving different transporters in a cell-specific manner. Compensatory mechanisms in response to SLC1A5 knockdown deserve further investigation as it may have implications in strategies aiming at inhibiting its activity. Further expression, localization and functional studies will follow to better understand changes occurring in lung cancer progression.
Key words: Non-small cell lung cancer, SLC1A5, expression, amino acid, transporters
Citation Format: Pierre P. Massion, Maria Senosain, Mohamed Hassanein, Xiangming Ji, Jun Qian, Megan hoeksema. Regulation of neutral amino acid transporters gene expression profile in non-small cell lung cancer. abstract. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1185. doi:10.1158/1538-7445.AM2015-1185
Lung cancer is the leading cause of death among all the cancers. Non‐small cell lung cancer accounts for 80% of lung cancer with a five‐year survival rate of 16%. Notch pathway, especially Notch‐1 is ...upregulated in a subgroup of Non‐small cell lung cancer patients. Since Notch‐1 signaling plays an important role in cell proliferation, differentiation, and apoptosis, down‐regulation of Notch‐1 may exert anti‐tumor effects. The objective of this study was to investigate the effect of delta‐tocotrienol on non‐small cell lung cancer cell lines (A549, H1299) via Notch signaling. Treatment with delta‐tocotrienol resulted in a dose and time dependent inhibition of cell growth (p<0.05), cell migration, tumor cell invasiveness, and induction of apoptosis (p<0.05). Western blot analysis showed that apoptosis induced by delta‐tocotrienol was associated with decrease in Notch‐1, Hes‐1, Survivin, and BCX‐XL expression. This was confirmed by real‐time PCR by a decrease in the gene expression of Notch1, Hes‐1, MMP‐9, and VEGF was observed. In addition, this was accompanied with a decrease in NF‐κB‐DNA binding activity. These results suggest that down‐regulation of Notch‐1, especially by delta‐tocotrienol could provide a novel approach for the prevention of tumor progression, which is likely to be mediated via downregulation of NF‐κB signaling pathways in Non‐small cell lung cancer.
Grant Funding Source: Across Societies (EB)
Abstract
The amplification of the distal portion of chromosome 3q in lung cancer is a major signature of neoplastic transformation, particularly in the squamous cell carcinoma (SCC) of the lung. A ...number of potential drivers in the 3q amplicon have been identified and are proposed to contribute to the development of lung SCC. miRNAs have emerged as a new class of small, non-coding RNAs that regulate gene expression and might act as integral parts of the molecular architecture of oncogene and tumor suppressor networks in human cancer, including non-small cell lung cancer (NSCLC). miR 1224-5p, 1248 and 944 are located at 3q amplicon and overexpressed in 50%, 26% and 53% of 30 primary SCCs when compared to normal tissues. The overexpression of these miRNAs was further validated in 237 TCGA lung SCC samples compared with 35 normal tissues (p=0.007, 1.18E-16 and 7.71E-20, respectively). The expression of the three miRNAs was higher in lung SCC compared to lung adenocarcinomas (p=8.19E-05, 1.04E-33 and 3.37E-95, respectively), suggesting unique roles of these miRNAs in the lung SCC development. Using miRNA mRNA target prediction tools (miRANDA and TargetScan) and in the combination with a literature search, we found RUNX1T1 and SMAD4 were candidate tumor suppressor gene target for miRNA-1224-5p and miRNA-1248. Downregulation of RUNX1T1 and SMAD4 were confirmed in TCGA lung SCC dataset. Using real-time RT-PCR, we found that the same three miRNAs 1224-5p, 1248 and 944 are overexpressed in 3q amplified lung SCC cell lines H520 and HCC95. When miRNA inhibitors were used to repress the expression of miR-1224-5p or miR-1248 in H520, we found that RUNX1T1 and SMAD4 protein expression levels were increased, suggesting that these tumor suppressor genes are regulated by miR-1224-5p or miR-1248. Lastly, knockdown of miR-1224-5p or miR-1248 using miRNA inhibitors led to 35-60% growth inhibition in H520 and HCC95 cells. Together, these results indicate potential roles for miR-1245-5p and 1248 in regulating cell proliferation by directly targeting tumor suppressor genes RUNX1T1 or SMAD4 in lung SCC. This work provides, for the first time, evidence for overexpression of miRNAs at the 3q26-29 amplicon that might be implicated in the oncogenesis of lung SCC. The work is supported by the NIH Grant R01 CA102353 to PPM and Lung Cancer Research Foundation grant (2011) to JQ.
Note: This abstract was not presented at the meeting.
Citation Format: Jun Qian, Xiangming Ji, Yong Zou, Megan Hoeksema, Heidi Chen, Pierre P. Massion. Overexpression of chromosome 3q26-29 miRNAs in squamous cell carcinoma of the lung. abstract. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 526. doi:10.1158/1538-7445.AM2014-526
...COPD variants may have pleiotropic effects that influence multiple traits. Other phenotypes in the MyCode dataset associated with the six SNPs included lung cancer, COPD, rheumatoid arthritis, and ...hypothyroidism. Furthermore, in patients with emphysema, cigarette smoke activates the production of antielastin antibody and a T-helper cell type 1 immune response, which has a known role in mediating autoimmune diseases such as type 1 diabetes (12, 13). Supported by National Institutes of Health grants K07CA172294 (M.C.A.) and RO1 LM0010685 (J.C.D.), Department of Defense grant W81XWH-11-20161 (P.P.M.), and Flight Attendant Medical Research Institute grant YFEL141014 (X.J.).
Chronic obstructive pulmonary disease (COPD) remains a leading cause of death and treatment options for this condition remain scant. This study aims to examine the ability of γ-tocotrienol to ...mitigate disease progression in an animal model of COPD induced by e-cigarettes. We hypothesize that γ-tocotrienol will attenuate inflammation and subsequently slow the progression of e-cigarette induced COPD.
Scnn1b-Tg + mice (n = 10/group), were exposed to e-cigarette vapor twice daily for 10 days in an acute model and daily for eight weeks in a chronic model with or without γ-tocotrienol at a dose of 100 mg/kg/day. Following the treatment, animals underwent pulmonary function testing. Upon sacrifice, bronchoalveolar lavage (BAL) fluid and serum were collected for analysis of cytokine expression through cytokine array. Markers of oxidative stress, inflammation, and fibrosis in the lung were assessed via western blot. Mucus accumulation and structural changes (i.e., emphysema) were measured through Periodic Acid-Schiff and Hematoxylin and Eosin staining, respectively. Collagen deposition were evaluated by Sirius Red staining and Sircol Collagen assay. Cell morphology in BAL fluid was analyzed by Diff staining.
In the acute model, γ-tocotrienol was evidenced to decrease collagen deposition and mucus accumulation in the bronchioles. Additionally, γ-tocotrienol reduced expression of cytokines C-X3-C motif chemokine ligand (CX3CL) 1 (P = 0.017), interleukin (IL) 4 (P = 0.0038) and T-cell immunoglobulin and mucin domain (TIM) 1 (P = 0.0215). Evidenced by large effect size, pulmonary function tests evidenced the ability of γ-tocotrienol to preserve lung function following 8 weeks of e-cigarette exposure. In addition, macrophage presence in BAL fluid was decreased following in mice supplemented with γ-tocotrienol following e-cigarette exposure.
Our results show the ability of γ-tocotrienol to attenuate the inflammatory response, and preserve lung function in models of e-cigarette induced COPD. These results indicate potential beneficial effects of γ-tocotrienol as an ancillary treatment in COPD.
This work was supported by FAMRI foundation “METABOLIC REPROGRAMMING IN PATIENTS WITH COPD”, YCSA 2015.
Plasma HDL cholesterol levels are inversely correlated with cardiovascular disease, the leading cause of death worldwide. This study investigated the effect of algae extract (AE) and its ...sub‐fractions (F1,2,3,4) on plasma lipid concentrations in hamster model. 80 male golden Syrian hamsters were randomized into: control(C)/water (W); C/AE; high fat diet (HF)/W; HF/AE; HF/F1; HF/F2; HF/F3 and HF/F4(n=10). AE and its subfractions (20%) were administrated via the drinking water for 4 weeks. Serum lipid profile was assayed and liver samples were subjected to rT‐PCR for the relative transcription levels of genes involved in HDL/reverse cholesterol transport (RCT) metabolism (Apo‐A1, ABCA1, CETP, and SRB‐1). Significant reduction was seen in TC (F3, p<0.05) and in non‐HDLc (AE, p<0.05), (F3, F4, p<0.01) as compared to HF/W, while the HDLc showed significant increases in the AE (p<0.05), F3 and F4 (p<0.01) groups. qPCR showed a 2 fold increase in ApoA1 and ABCA1 expression(F4) indicating an increase in HDL production, biogenesis, and maturation. Increase in SRB1 expression (2 fold) indicates that F4 further augments RCT mechanism. Reduction of CETP expression may indicate a decrease in transfer of CE to LDL, further increasing cholesterol held as HDL particles. Algal extracts and/or its sub‐fractions significantly improved plasma cholesterol profile by lowering LDL and increasing HDL, possibly via the RCT mechanism
Grant Funding Source: Health Enhancement Product Inc.
Isoprenoids suppress the mevalonate pathway that provides prenyl groups for the posttranslational modification of growth-regulating proteins. We hypothesize that xanthorrhizol and
-δ-tocotrienol ...synergistically suppress the growth of murine B16 melanoma and human DU145 prostate carcinoma cells. Xanthorrhizol (0-200 µmol/L; half maximal inhibitory concentration IC
= 65 µmol/L) and
-δ-tocotrienol (0-40 µmol/L; IC
= 20 µmol/L) each induced a concentration-dependent suppression of the proliferation of B16 cells and concurrent cell cycle arrest at the G1 phase. A blend of 16.25 µmol/L xanthorrhizol and 10 µmol/L
-δ-tocotrienol suppressed B16 cell proliferation by 69%, an impact greater than the sum of those induced by xanthorrhizol (15%) and
-δ-tocotrienol (12%) individually. The blend cumulatively reduced the levels of cyclin-dependent kinase four and cyclin D1, key regulators of cell cycle progression at the G1 phase. The expression of RAS and extracellular signal-regulated kinase (ERK1/2) in the proliferation-stimulating RAS-RAF-MEK-ERK pathway was downregulated by the blend. Xanthorrhizol also induced a concentration-dependent suppression of the proliferation of DU145 cells with concomitant morphological changes. Isobologram confirmed the synergistic effect of xanthorrhizol and
-δ-tocotrienol on DU145 cell proliferation with combination index values ranging 0.61-0.94. Novel combinations of isoprenoids with synergistic actions may offer effective approaches in cancer prevention and therapy.
Xanthorrhizol, a sesquiterpene, and d-δ-tocotrienol, a vitamin E molecule, each suppresses the proliferation of a number of tumor cells. This study aims to examine the potentially synergistic effect ...of xanthorrhizol and d-δ-tocotrienol in tumor cells.
Murine B16 melanoma and human DU145 prostate carcinoma cells were incubated for 48 h (B16) or 72 h (DU145) with xanthorrhizol or d-δ-tocotrienol before cell populations were determined by CellTiter 96â Aqueous One Solution. Cells incubated with the agents for 24 hours were stained with propidium iodide and analyzed for cell cycle using flow cytometry and MultiCycle AV. Isobologram and combination index (CI) were used to demonstrate their synergistic anti-proliferative impacts.
Xanthorrhizol (0–200 μmol/L) and d-δ-tocotrienol (0–40 μmol/L) each elicited a concentration-dependent suppression of the proliferation of B16 cells. A blend of 16.25 μmol/L xanthorrhizol and 10 μmol/L d-δ-tocotrienol achieved 69% (P < 0.05) growth suppression of B16 cells, exceeding the sum of individual effects. B16 cells incubated with 5 and 10 μmol/L d-δ-tocotrienol for 24-h had a concentration-dependent increase in the percentage of cells in the G1 phase with a concomitant decrease in the percentage of cells in the S phase. The G1/S ratio, an indicator of cell cycle arrest at the G1 phase, increased from 1.73 ± 0.05 (Control) to 2.01 ± 0.10 (5 μmol/L) and 2.73 ± 0.05 (10 μmol/L). A parallel pattern of concentration-dependent increase in the G1/S ratio was induced by xanthorrhizol at concentrations equivalent to 25% (16.25 μmol/L) and 50% (32.5 μmol/L) of its IC50 value. A blend of 5 μmol/L d-δ-tocotrienol and 16.25 μmol/L xanthorrhizol, each at no-effect concentrations, significantly increased the percentage of B16 cells in the G1 phase to 62.6 ± 0.6%. Isobologram and CI confirmed the synergistic effect of xanthorrhizol (50 and 100 μmol/L) and d-δ-tocotrienol (10 and 20 μmol/L) on the proliferation of DU145 cells.
Xanthorrhizol and d-δ-tocotrienol synergistically suppress tumor cell proliferation by inducing G1 arrest and may have potential in cancer prevention and therapy.
American River Nutrition, Inc. and the University Assistantship Program and the Department of Nutrition of Georgia State University.
The molecular determinants of lung cancer risk remain largely unknown. Airway epithelial cells are prone to assault by risk factors and are considered to be the primary cell type involved in the ...field of cancerization. To investigate risk-associated changes in the bronchial epithelium proteome that may offer new insights into the molecular pathogenesis of lung cancer, proteins were identified in the airway epithelial cells of bronchial brushing specimens from risk-stratified individuals by shotgun proteomics. Differential expression of selected proteins was validated by parallel reaction monitoring mass spectrometry in an independent set of individual bronchial brushings. We identified 2,869 proteins, of which 312 proteins demonstrated a trend in expression. Pathway analysis revealed enrichment of carbohydrate metabolic enzymes in high-risk individuals. Glucose consumption and lactate production were increased in human bronchial epithelial BEAS2B cells treated with cigarette smoke condensate for 7 months. Increased lipid biosynthetic capacity and net reductive carboxylation were revealed by metabolic flux analyses of U-
C
glutamine in this in vitro model, suggesting profound metabolic reprogramming in the airway epithelium of high-risk individuals. These results provide a rationale for the development of potentially new chemopreventive strategies and selection of patients for surveillance programs.
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