bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A28: Loss of inhibitory IRS-1 phosphorylation is an early event in mTOR-dependent endometrial hyperplasia and carcinoma
Authors: McCampbell Adrienne , Harris Heather , Crabtree Judy , Winneker Richard , Walker Cheryl , Broaddus Russell
Published: May 2012

A28

Activation of IGF-I receptor signaling contributes to the development of endometrioid-type endometrial carcinoma in humans and in rodent models. This signaling pathway is under both positive and negative regulation, including S6K phosphorylation of IRS-1 at S636/639, which occurs downstream of mTOR activation to inhibit this adapter protein. We observed activation of mTOR signaling with a high frequency in human endometrial hyperplasia and carcinoma, but an absence of IRS1 phosphorylation at S636/639, despite the high levels of activated S6K in these tumors. To explore when during disease progression mTOR activation and loss of inhibitory IRS-1 phosphorylation occurred, we utilized the Eker rat (Tsc2Ek/+) model for this disease, where endometrial hyperplasia develops as a result of loss of Tsc2, the “gatekeeper” for mTOR signaling. We observed mTOR activation early in progression in hyperplasias and in some histologically normal appearing epithelial cells, suggesting that event(s) in addition to loss of Tsc2 were required for progression to hyperplasia. In contrast, IRS-1 phosphorylation at S636/639 was observed in normal appearing epithelium, but was absent from all hyperplasias, indicating loss of IRS-1 inhibition by S6K occurred during progression to hyperplasia. Furthermore, treatment of Eker rats with an mTOR inhibitor resulted in a decreased incidence of endometrial hyperplasia [54/79 hyperplasias/uterine cross section in vehicle controls to 6/90 in treated rats (p<0.001)] and decreased the proliferative index of hyperplastic endometrial epithelial cells by >95% (p < 0.05). Since progression from normal epithelium to endometrial carcinoma proceeds via the intermediate step of endometrial hyperplasia, these data suggest a progression sequence where activation of mTOR is followed by loss of negative feedback to IRS-1 during the initial stages of development of this disease.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A28.
Abstract A3: Bcl-w-induced invasive pathways in gastric cancer cells
Authors: Um Hong Duck , Bae In Hwa , Yoon Sung Hwan
Published: May 2012

A3

We have reported previously that Bcl-w, a pro-survival member ofthe Bcl-2 family, is expressed in gastric cancer cells, particularly in those with an infiltrative morphology. Functional studies utilizing Bcl-w-overexpressing cells have consistently demonstrated that Bcl-w enhances not only the survivability of the cancer cells, but also their migratory and invasive potentials. The Bcl-w-induced invasive pathway was determined to involve a sequential activation of phosphoinositide 3-kinase (PI3K), Akt, and Sp1, which subsequently results in the expression of metalloproteinase-2 (MMP-2). In this study, we have demonstrated that Bcl-w also induces the urokinase-type plasminogen activator(uPA), which was prevented by treatment with pharmacological inhibitors of PI3K, Akt, or Sp1. These inhibitors also attenuated the ability of Bcl-w to promote cell migration. Similar results were obtained when the actions of MMP-2 and uPA were blocked using their inhibitors or small interfering RNAs (siRNAs), thereby suggesting that these enzymes stimulate migratory signals in this system. Bcl-w overexpression was shown to activate focal adhesion kinase (FAK) in a manner dependent on PI3K, Akt, Sp1, MMP-2, and uPA, and the prevention of FAK action using its siRNAs or dominant negative mutants abolished Bcl-w-induced migration. Overall, it appears that Bcl-w induces both MMP-2 and uPA via the PI3K-Akt-Sp1 pathway, which then promotes cell invasion not only via the degradation of the extracellular matrix, but also via the stimulation of the FAK-dependent migratory pathway.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A3.
Abstract A30: MicroRNA-27a functions as an oncogene in gastric adenocarcinoma by targeting prohibitin
Authors: Tang Hua , Liu Tao , Lang Yuanyuan , Liu Min , Li Yixuan , Li Xin
Published: May 2012

A30

MicroRNAs (miRNAs) may function as oncogenes or tumor suppressors. Here, we show that miR-27a is up-regulated in human gastric adenocarcinoma. Suppression of miR-27a inhibits gastric cancer cell growth. Subsequently, prohibitin is identified as a potential miR-27a target, combining bioinformatics and microarray analysis. EGFP report experiment also confirms that the 3’ untranslated region (3’ UTR) of prohibitin carries the directly binding site of miR-27a. After knockdown of miR-27a in gastric cancer cells, mRNA level and protein level of prohibitin are both elevated. Down-regulation of prohibitin by miR-27a may explain why suppression ofmiR-27a can inhibit gastric cancer cell growth, further supporting that miR-27a functions as an oncogene.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A30.
Abstract A31: Intestinal resistance to celecoxib in Min/+ mice involves ECM changes and an expanded population of TGFβ-expressing myofibroblasts
Authors: Davids Jennifer , Carothers Adelaide , Damas Beatrice , Bertagnolli Monica
Published: May 2012

A31

The COX-2 inhibitor, celecoxib, inhibits prostaglandin E2 (PGE2) production and intestinal adenoma formation in mice and in humans. Previously, we showed that treatment of Min/+ mice with dietary celecoxib for 3 weeks reduced PGE2 and induced tumor regression, whereas treatment for 5 months resulted in increased PGE2 and tumor growth (Carothers et al. Cancer Res. 2006; 66: 6432-8). In the small intestine, cross-talk between epithelial and stromal cells occurs across the extracellular matrix (ECM). Heparan sulfate proteoglycans (HSPGs) within the ECM sequester and modulate the activities of growth factors and latent TGFβ ligands. TGFβ stimulates differentiation of fibroblasts into PGE2-producing myofibroblasts. These stromal cells play important roles in intestinal tumorigenesis and in the chemopreventive effect of nonsteroidal anti-inflammatory drugs. We tested the hypothesis that celecoxib resistance in the intestine of Min/+ mice involves ECM regulation of HSPGs and TGFβ signaling in epithelial and stromal cells. Using immunohistochemistry (IHC), we compared the non-tumor mucosa and adenomas of untreated Min/+ controls with Min/+ mice treated with celecoxib for 3 weeks and for 5 months. We performed IHC for the HSPG membrane protein syndecan-1, cytokines TGFβ1, 2, 3 (TGFβ), and the nuclear downstream TGFβ effector, Smad4. IHC was also performed to characterize subepithelial stromal cells using F4/80, a macrophage-specific marker, as well as α-smooth muscle actin (αSMA) and vimentin, myofibroblast-specific markers. In comparison to untreated Min/+ small intestine, celecoxib treatment for 3 weeks increased membrane-localized syndecan-1, TGFβ, and nuclear Smad4 in crypt and villus enterocytes. Under these conditions, few αSMA+ and vimentin+ myofibroblasts or F4/80+ macrophages were present in the pericryptal submucosa and lamina propria. Thus, brief treatment may sequester latent TGFβ and growth factors in the ECM, blocking their downstream signaling. A reciprocal effect was produced by 5 month treatment, in which there was markedly reduced expression of membrane-localized syndecan-1, TGFβ, and nuclear Smad4 in the crypt and villus enterocytes. There was also a dramatic increase in the number of myofibroblasts in the lamina propria and pericryptal submucosa (P<.01). Thus, celecoxib resistance was associated with TGFβ-induced differentiation of fibroblasts into PGE2-producing myofibroblasts. These results suggest that the duration of celecoxib treatment affects the cross-talk between epithelial and stromal cells through the ECM, providing a novel mechanism for celecoxib resistance. Moreover, elevated numbers of myofibroblasts in the human colon may be predictive of chemoprevention failure.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A31.
Abstract A33: Superior efficacy of dietary 3,3’diindolylmethane (DIM) amongst several indoles to inhibit prostate cancer growth via induction of the p75NTR tumor suppressor protein
Authors: Khwaja Fatima , Posey Isadora , Song Xueqing , Wynne Shehla , Djakiew Daniel
Published: May 2012

A33

A comparison of the indoles etodolac, indomethacin, 5-methylindole-3-acetic acid, indole-3-carbinol, 3,3’diindolylmethane (DIM) and the related ketorolac molecule showed superior efficacy of DIM, a dietary indole, to induce expression of the p75NTR tumor suppressor and inhibit survival of the PC-3 prostate cancer cell line. Each indole that induced p75NTR expression exhibited the same rank-order for inhibition of prostate cell survival. Comparison of the activity of DIM, the most potent of the indoles, between several cell lines, showed that DIM induced p75NTR at low concentrations in prostate cells (PC-3 , DU-145), followed by bladder (T24), with breast (MCF-7) and fibroblast cells (3T3) being the least responsive to DIM. DIM induction of p75NTR levels in the various cell lines occurred in the same rank-order as inhibition of cell survival. Since the consumption of cruciferous vegetables has been reported to reduce the risk of prostate cancer through the release of indole-3-carbinol (I3C) which is subsequently metabolized in the acidic environment of the stomach to DIM, the superior activity of DIM to induce expression of the p75NTR tumor suppressor protein and inhibit cell survival suggests a mechanism of action for the anticancer activity of DIM in the prevention of prostate cancer. In order to demonstrate a cause and effect between DIM and p75NTR inhibition of survival, we used a dominant negative of p75NTR to rescue DIM induced loss of cell survival. In order to investigate the mechanism of action by which p75NTR transduces a biochemical signaling cascade leading to cell death of prostate cancer cells we utilized knockdown with p38 MAPK siRNA to rescue DIM induction of p75NTR thereby implicating p38 MAPK in DIM induction of cell death. Moreover, DIM treatment induced phosphorylation of p38 MAPK within one minute, suggesting the target of DIM is proximal to p38 MAPK. Hence, these results suggest that the anti-cancer activity of cruciferous vegetables may be mediated in part by DIM and that the mechanism of action occurs, in part, by activation of the p38 MAPK pathway leading to expression of the p75NTR death receptor with subsequent apoptosis of prostate cancer cells.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A33.
Abstract A38: Structure-activity relationship of closely-related flavonoids with possible chemopreventive qualities
Authors: Tsuji Petra , Stephenson Katherine , Wade Kristina , Liu Hua , Fahey Jed
Published: May 2012

A38

Dietary flavonoids are now widely presumed to have human health benefits, and increased intake is epidemiologically correlated with decreased risk of cardiovascular disease and cancer. The general mechanisms thought to be involved in chemoprevention through dietary compounds include the inhibition of the carcinogen-bioactivating phase I enzymes and/or induction of carcinogen-bioinactivating phase II enzymes. Methoxylated flavonoids have been shown to have chemopreventive activity in a number of in vitro assays, and we have selected 27 of them as the objects of this work. Furthermore, these flavonoids were analyzed to investigate possible structure-activity relationships for effective, non-toxic agents with chemopreventive potential.

The flavonoids are a structurally diverse class of low molecular weight polyphenolic benzo-c-pyrone derivatives. The basic structure is composed of two benzene rings (A and B) separated by an oxygen-containing heterocyclic ring. Substituent groups can attach on each of the molecule’s rings, with hydroxyl, methoxy groups or sugar moieties being present most frequently. Neither hydroxylation nor methoxylation solely on the B-ring had a strong inducing effect on the phase II enzyme NAD(P)H:quinone-reductase (NQO1) in murine hepatoma cells. However, flavonoids with a methoxy-group substitution at the 5-position of the A-ring were the most potent inducers of NQO1. Other flavonoids were equally potent inducers, but exhibited much higher toxicity in at least one of the cell lines tested. Certain methoxylated flavonoids that are found in dietary fruits and vegetables have significant potential for prevention of chemically-induced carcinogenesis and should be investigated further in animal models of cancer prevention.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A38.
Abstract A39: Dosage-dependent regulation of caspase 8 activation in LNCaP and DU-145 human prostate cancer cell lines by the Chinese medicinal herb scutellaria barbata.
Authors: Nguyen Dinh , Devadhason Rachel , Wong Brian
Published: May 2012

A39

Scutellaria barbata (SB) (known in traditional Chinese medicine as Ban-Zhi-Lian) is a perennial herb. It has been used to treat appendicitis, hepatitis, pulmonary abscess, and several cancers. LNCaP cells are derived from a human prostate cancer cell line with mutated androgen receptors (AR) while DU-145 is a classical human prostate cell line without androgen receptors. Our previous data indicated that SB activated apoptosis in LNCaP cells via caspases 3, 8, and 9. Histopathological data revealed that the herb had the ability to delay tumor progression and development in TRAMP mice. Recent Western blot data from LNCaP cells undergoing treatment with SB showed increased expression of Akt protein while decreased phosphorylation of Akt in the cells. In this study, we compared the effect of AR-dependence on the herb's apoptosis activation through caspase 8 on these two different cell lines. The two cell lines were incubated with distilled water as a control group and 100 µL and 200µL of SB as experimental groups for 2 and 8 hours separately. A specific carboxyfluorescein (FAM) labeled peptide fluoromethyl ketone (FMK) caspase inhibitor (FAM-Peptide-FMK) was used to label caspase 8- activated cells. Caspase positive (+) cells appeared green whereas necrotic cells were stained red with propidium iodide and observed under a fluorescent microscope. Pictures were taken and the activation/necrosis ratio (A/N ratio) was analyzed using ImageJ software (developed by the National Institutes of Health). Significant induction of caspase 8 by SB was observed in androgen receptor positive LNCaP cells. In both 2 and 8 hour incubation, SB activated more caspase 8 in LNCaP cells than in DU-145 cells treated at 100 µL and 200 µL concentrations ( A/NLNCaP > A/NDU-145 ). In addition, DU-145 exhibited significant dosage-dependent induction of caspase 8 by SB, while dosage-dependence was not observed in LNCaP cells (A/N100µL = A/N200µL). These data indicate that SB might activate apoptosis through an up-stream androgen receptor pathway, thus suggesting a plausible mechanism for this herb’s activation of the apoptotic pathway in prostate cancer cells.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A39.
Abstract A4: Antagonistic effects of sulphated polysaccharides from Turbinaria conoides (J. Agardh) on tumor cell migration and angiogenesis
Authors: Delma Caroline , Ramalingam Karthikeyan , Pandian Vijayabaskar , Baskar Albert , Savarimuthu Ignacimuthu , Thangavelu Balasubramanian , Somasundaram Somasundaram
Published: May 2012

A4

Seaweeds are considered to be a rich source of biologically active substances like antioxidants, anti-viral, anti-inflammatory and anticoagulant agents. There are some evidences that seaweeds contain compounds with a relatively high antioxidant and antiproliferative activity. Seaweeds are low in fats but contain vitamins and bioactive compounds, like terpenoids and sulfated polysaccharides. Sulphated polysaccharides from brown algae are found to exhibit diverse biological activities like anticoagulant, antihyperlipidemic, antiviral, antioxidant, antitumor, antimetastatic and antiangiogenic activities. In this study, the antiproliferative, antiinvasive and antiangiogenic properties of sulphated polysaccharides from brown seaweed, Turbinaria conoides against colon carcinoma cell line COLO 320DM were investigated. Crude sulphated polysaccharide was extracted from T. conoides, collected from Mandapam coast, South east coast of India. The antiproliferative activity of the seaweed polysaccahrides on COLO 320DM was evaluated by MTT assay and DNA fragmentation studies. The inhibitory potential on the migration of this invasive cell line was assessed by wound healing assay. The effect of sulphated polysaccharides against angiogenesis was investigated by chorio allantoic membrane (CAM) assay in chick embryo. It was observed that sulphated polysaccharides from T. conoides inhibited the proliferation of COLO 320DM cells at a concentration of 1mg/ml by 40% at 24 hours. DNA fragmentation as evidenced by ladder formation in agarose gel indicates that the antiproliferative activity of seaweed polysaccharides is due to the induction of apoptosis. The migration of the carcinoma cell line was effectively inhibited by the seaweed extract. The antiinvasive property increased with increasing concentrations. However, the activity was more or less similar at 24 and 48 hours. In the CAM assay, it was found that sulphated polysaccharides from the brown seaweed inhibited vasculogenesis in a developing chick embryo. These results clearly indicate that sulphated polysaccharides from brown algae are potential molecular leads for antimetastatic drug development.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A4.
Abstract A40: The enhancement of p53 stability via the inhibition of JNK-modulated ubiquitination with the activation of redox factor 1 (Ref-1) in response to non-genotoxic antioxidant selenomethionine (semet)
Authors: Jung Hwa Jin , Lee Ju Han , Jeong Seok Won , Kwon Jee Young , Park Jee Yeon , Kim Hye Lim , Lee Young Ju , Chang Sung-Goo , Seo Young Rok
Published: May 2012

A40

p53 has been known to play an important role as an tumor suppressor gene for the maintaining of genome stability. In the field of cancer prevention study, p53 has been issued to clarify the mechanism of activation under the non-genotoxic cancer preventive agents containing antioxidant selenium. In previous our study, p53 has been reported to be the function of transcriptional activation activated without the genotoxicity through the redox modulation in response to selenomethionine (SeMet). In this study, we investigated that the mechanism of p53 protein stabilization enhanced by SeMet. Our data showed that the ubiquitinated p53 was decreased in the cells exhibiting the downregulation of JNK which is one of E3-ubiquitin ligases compared with in mock-treated cells suggesting that the JNK-modulated p53 ubiquitination might be inhibited by SeMet. To define the mechanism of the enhanced p53 stability in the presence of SeMet, redox factor 1 (Ref-1) which has been documented as one of the factors modulating the redox status in response to antioxidants was downregulated using siRNA. Our data showed the first time that the interaction of JNK and p53 was increased in Ref-1 siRNA-treated cells in the presence of SeMet implicating that the Ref-1 activated by SeMet might be involved in the inhibition of JNK-mediated p53 ubiquitination. Our study suggested that the enhancement of p53 stabilization in response to SeMet might provide an important clue for the cancer prevention.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A40.
Abstract A44: Carnosol, a dietary diterpene functions as a dual AR and ER-alpha antagonist in prostate cancer.
Authors: Johnson Jeremy , Syed Deeba , Heren Chenelle , Siddiqui Imtiaz , Mukhtar Hasan
Published: May 2012

A44

Emerging evidence suggest that androgens may not be the only hormone responsible for the etiology of prostate cancer (PCa). Recent data has suggested that estrogens may be another important piece to the PCa puzzle. For example the use of the Selective Estrogen Receptor Modulator (SERM) toremifene in HG-PIN patients for one year resulted in a 21% reduction in the conversion of HG-PIN to PCa after one year of treatment. It is well established that race correlates with the levels of androgens with African Americans having the highest, followed by Caucasians and lowest in Japanese males. A similar trend has been observed in regard to serum estrogen levels and it is noteworthy that with age estrogen levels remain constant while androgens levels decrease. Given that experimental animal models suggest that both estrogens and androgens are necessary for PCa development it may be a reasonable approach to simultaneously disrupt both androgen and estrogen signaling in PCa. Several SERMs that include tamoxifen, toremifine, and fulvestrant have been evaluated for their ability to also target the AR and function as a dual AR and ER antagonist, however, experiments have suggested these effects are saturable and in some instances SERMs exert AR agonist properties. In our quest to find a dual AR and ER antagonist present in dietary substances that humans consume we found that the dietary diterpene Carnosol displays growth inhibitory properties in the hormone sensitive prostate (LNCaP and CWR22Rv1) and breast (MCF7 and AU565) cancer cell lines. Given the structural similarity between Carnosol to dihydrotestosterone and estradiol we hypothesized that it may interact with both the androgen and estrogen receptor alpha (ER-α). Using a TR-FRET cell free biochemical assay we found that Carnosol can interact with both AR and ER-alpha with similar potencies. Next, we evaluated the antagonist and agonist properties of Carnosol using a functional assay where the AR and ER-alpha ligand binding domains are expressed as a fusion protein in HEK293 cells. We observed that Carnosol exhibits antagonist properties that are dose responsive and even more interestingly, regardless of dose, did not exert agonist properties. In LNCaP, CWR22Rv1, and MCF7 cells Carnosol was found to disrupt both androgen and estrogen signaling in a dose and time dependent manner. Our finding that Carnosol displays the ability to disrupt both androgen and estrogen signaling simultaneously suggesting that Carnosol may be developed or chemically modified through more rigorous structure activity relationship studies of other such chemical entities for a new class of investigational agents - a dual AR/ER modulator.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A44.
Abstract A46: Ablack raspberry extract inhibits the proliferation of cervical cancer cells partially through proapoptotic effects
Authors: Zhang Zhaoxia , Seamon Leigh , Knobloch Thomas , Stoner Gary , Cohn David , O'Malley David , Paskett Electra , Fowler Jeffrey , Weghorst Christopher
Published: May 2012

A46

Cervical carcinoma, the second most common female cancer worldwide, is the seventh leading cause of cancer death in women. Although pap smears to screen abnormal cervical cytology have helped reduce mortality rates, it remains a challenge to manage preinvasive and invasive cervical lesions. The long latency of cervical cancer allows preventive and therapeutic interventions before progressing onto invasive disease. Thus, phytochemical-based cancer chemoprevention and the role of bioactive food components have attracted significant interest in current cancer-fighting interventions. We have previously shown that lyophilized black raspberries (LBR, Rubus occidentalis) inhibit the cell proliferation of human oral squamous cell carcinoma cells in vitro as well as oral tumorigenesis in vivo. Given the similar epidemiologic basis of oral and cervical cancers, we evaluated whether an LBR extract had anti-proliferative effects in cervical cancer cells in vitro and began to elucidate the possible regulatory molecular mechanisms. We found that LBR inhibited the proliferation of three human cervical cancer cell lines, HeLa (HPV16−/HPV18+) SiHa (HPV16+/HPV18−) and C-33A (HPV18−/HPV16−), in a dose-dependent manner (at 25, 50, 100 or 200µg/ml for 1, 3 and 5 days, respectively) to a maximum of 44%, 53% and 45% respectively (P<0.005). While exogenous hydrogen peroxide (H2O2, 100µM), a common reactive oxygen species (ROS) intermediate and inhibitor of cell growth, inhibited C-33A proliferation, HeLa and SiHa cells were not significantly affected. The anti-proliferative effects of either LBR or LBR+catalase were similar in all cell lines as shown by WST1 (tetrazolium salt cleavage) and crystal violet assays, which supporting the hypothesis that decreased proliferation is specific to LBR bioactive components and independent in vitro peroxide generation. Flow cytometric analysis of propidium iodide and Annexin V staining showed that LBR induced apoptotic markers in all three cells lines, and did not change cell cycle progression. LBR treatment induced hypercondensation and volume contracted nuclei in Hela cells as demonstrated by fluorescence microscopy analysis of Hoechst 33342 staining. In conclusion, an extract of lyophilized black raspberries demonstrated significant anti-proliferative and pro-apoptotic activities in human cervical cancer cells in vitro. Therefore, black raspberries and their bioactive components may represent promising candidates for the phytochemical-based chemoprevention of cervical cancer and warrant further investigation.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A46.
Abstract A51: Chemopreventive effects of Brazilian herb oral feeding on UVB induced photocarcinogenesis
Authors: Tokuda Harukuni , Kaneko Masafumi , Yamashita Mitsuaki , Iiida Akira
Published: May 2012

A51

Ultraviolet(UV) light is the most common cause of skin

cancer in humans and it is very acute problem in our life

style during living. Several effects of UVB(290-320 nm) are thought to contribute to skin carcinogenesis and generation of free radicals and related oxidants produced by UVB exposure.Tabebuia avellanedae (Bignoniaceae)(TA), which is native in South America from Brazil to northern Argentina, is well known in traditionalfolk medicne used for the treatment of various disease during five hundred years. The inner bark of this plant produced in Brazil is distributed in Asia and Japan as a healthy purpose herb tea. Previously, we reported that extract essense of TA(TA ess.) and including naphthoquinones type compounds, NQ801, inhibited TPA-induced in vitro assay for chemopreventive potency and also inhibited TPA-induced in vivo assay on two-stage mouse skin test. We have now extended these investigations to a new tumorigenesis model in which we initiated the tumors with UVB irradiation and promoted with 1.7 nmol of TPA in SENCAR mice. These tumor -free mice, which had a high risk of developing skin tumors, using these experimental systems, were then treated drinking with TA ess. Oral feeding of 0.0025 % of TA ess., two weeks before and after tumor initiation resulted in a significant reduction in tumor incidence (40 %) accompanied by an extension(> 20 %) of the tumor latency and also decrease papilloma multiplicity. These results provide a basis for further development of TA ess. against sunlight-induced skin cancer in humans.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A51.
Abstract A53: Apoptosis initiated in colon cancer cells when treated with muscadine grape extract
Authors: Hannah Lane , Brown Vondina , Ranwala Damayanthi , Wargovich Michael
Published: May 2012

A53

Research suggests strong correlations between the inflammation process and colon cancer progression, making it an attractive target for anti-inflammatory drugs and compounds. Muscadines (Vitis rotundifolia), a grape species native to the Southeastern United States, are rich in polyphenols containing many biologically important flavonoids that may have potential anti-inflammatory and anti-oxidant properties. Current research has shown that the muscadine grape has a higher total phenolic and flavonoid content than commercially available red grapes. A hallmark of progressive colon cancers is disruption of different signaling pathways leading to increased proliferation and escape from apoptotic mechanisms. We hypothesize that extracts containing high levels of biologically important polyphenols from the muscadine pomace would induce cell cycle arrest and apoptosis in HT-29 and HT-15 colon cancer cell lines. Polyphenolic compounds were extracted from muscadine pomace using 80% methanol in 6N hydrochloric acid at 60ºC. Whole concentrated extract (WCE) was fractionized using an Oasis HLB column to yield three different muscadine fractions (MF). We compared the cytotoxic effects of both cell lines when treated with WCE versus three different muscadine fractions (MF-1, MF-2, and MF-3). Cell proliferation assays showed a decrease in viable cell proliferation in both cell lines treated with MF-3. Apoptosis was confirmed through the use of flow cytometry and we found evidence of G1 arrest in MF-3 treated cells. Immunoblots for apoptosis-related proteins suggest apoptotic activation of the caspase-3 pathway in treated cells compared to non-treated cells. We conclude that extracts of muscadine grape contain biologically active polyphenols that inhibit the growth of HT-29 and HT-15 colon cancer cells via an apoptotic pathway.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A53.
Abstract A55: A screening method for identifying potential angiogenesis inhibitors
Authors: Caldwell Kelli , Charlton Shirley , Krill Diane
Published: May 2012

A55

Objective

Plants produce chemical compounds to prevent encroachment by competitors. These turf wars that generate defense products in plants have potential medicinal value for humans. For example, compounds with anti-angiogenic properties are useful in combating cancer by preventing new blood vessel formation to support the tumor. The search for new angiogenesis inhibitors is ongoing, and utilizes a variety of time-intensive cell culture models. In this report we introduce a rapid, inexpensive method for screening potential anti-angiogenic compounds in a model system that stimulates the production of secondary defense chemicals in plants, thus increasing the potential yield of effective compounds.

Plant chemicals under investigation for anti-angiogenic activity include flavonoids, a broad group of compounds that are present in most photosynthesizing cells of plants. Flavonoids share a common phenyl-chromanone structure (C6-C3-C6), and include thousands of compounds, subdivided into 6 major categories; flavones, flavonols, isoflavones, flavanols, flavanones, and anthocyanins. Humans do not synthesize flavonoids, but derive an average of 1 gram per day in their diet. Their reported pharmacological properties include antioxidative, anti-inflammatory and antiproliferative properties.

Methods

In order to induce defense chemical production, we chose thyme and mint plants to combine because thyme was observed to inhibit the spread of mint, a highly invasive vascular plant. The thyme plants grown alone, as well as the mint alone, were analyzed as controls. The plant leaves and stems were extracted and the flavonoid products were analyzed by reverse phase HPLC. The flavonoids that appear in the thyme in the presence of mint, that are not present in the thyme plants grown alone, were considered induced by the presence of mint. The induced compounds were collected and tested in a vertical plate assay that measures root length as a quantitative assay for drug sensitivity. The compounds were applied to the vascular root tissue of mint seedlings 5-7 days after germination.

Results

Of eight Thymus vulgaris compounds that were induced by the presence of mint, 5 were tested by bioassay, and one was found to have an inhibitory effect. Known angiogenesis inhibitors, VEGF receptor tyrosine kinase and amelioride hydrochloride, significantly reduced root growth compared to the control. An HPLC-isolated compound b, which eluted at 15.2 minutes, reduced the growth of root vascular tissue compared to the control and vehicle control, and 50% as well as known angiogenesis inhibitors.

Conclusion

This report demonstrates that induced secondary defense chemicals produced by thyme plants include flavonoid compounds. Useful inhibitors may be identified with this rapid, sensitive, and inexpensive technique.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A55.
Abstract A56: Alterations in female reproductive organs of cyclic rats treated with aqueous extract of moringa oleifera lam: Indication of possible role in epithelial ovarian cancer
Authors: Bose Chinmoy
Published: May 2012

A56

Background

A hormonal etiology of epithelial ovarian cancer has long been suspected seeing its incidence menopausal age, and now the role of FSHR has also been demonstrated. Many ovarian cancer cell line express FSHR in them. In studies of the anticancer potential of plants used in folk medicine of Bengal, extracts of plants such as Oroxylum indicum, Moringa oleifera lam, Aegles marmelos could be considered as potential sources of anticancer compounds. Amongst them only Moringa oleifera lam has unique anticancer as well as hormonal property, which may or may not be attributable to isothiocyanate or glucosinolate that it contains. An animal experiment was planned to see effect of Moringa root extract in female reproductive system of mice.

Methods

5 adult female mice of swiss strain of 30 gm each 2-control, 3-treated kept on stock diet, pellet, having nutritional value of 7 days. An aqueous extract of the root was prepared according to a traditional method. In brief, 500 g of the root were placed in a container with 750 ml of water and boiled for 30 min. The preparation was left standing to cool and was then filtered. The filtrate, containing 66.7 mg root in 1 mL, was placed in small vials and kept in 4 degree C refrigerator until use. 1 ml of extract was used orally daily for 45 days.

Results

Attenuation/shrinkage of ovary and uterus was seen while mice tolerated the herb extract well. There was reversal to pre estrus phase of adult mice as was revealed by PAP smear from vagina. In histology there was absence of follicles in comparison to control ovary. There was lesser amount of fibrosis in treated ovary.

Conclusion

Isothiocyanate of Moringa may inhibit proliferation of ovarian granulosa and other cells as it induces apoptosis via caspase-9 and -3 pathways, a family of calcium-dependent cysteine proteases. It may also act by inhibiting ERK1/2 and Akt survival signaling while simultaneously activating pro-apoptotic p38 and JNK1/2. There are reports to show that Moringa induced decrease in cerebral dopamine and norepinephrine which may influence and lower NGF and FSHR through central mechanisms. There is strong possibilty of using this agent in epithelial ovarian cancer and, as such, a cell line experiment is urgently necessary.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A56.
Abstract A57: Lipid extract from the ostrich fern Matteucia struthiopteris displays potent anti-cancer effects in microtumors from human malignant cell lines via disruption of actin cytoskeleton
Authors: Crawford Sarah , Penarreta Rafaella , Diamond Deanna
Published: May 2012

A57

Previous studies from the Cancer Biology Research Laboratory identified primitive plants of moss, fern and lichen species with anti-cancer activities. Current research involves the extraction of a lipid-containing fraction from the ostrich fern, Matteucia struthiopteris, with 1,000-fold greater anti-cancer activity than whole plant aqueous homogenates in pre-clinical assays of microtumors prepared from several solid tumor malignancies, including drug-resistant glioblastoma multiforme. Confocal laser microscopy of phalloidin-stained tumor cells demonstrated that a primary effect of the plant extracts is the disruption of cortical actin cytoskeleton in the cell membrane. This represents a previously unrecognized therapeutic target for anti-cancer agents.

The targeted actin-cytoskeleton may also comprise a novel chemopreventive target. A novel biophysical model, proposed by Crawford, suggests that the active lipid fraction may affect cytoskeletal integrity as a consequence of the interaction of plant isoprenoids and dolichols on tumor membrane fluidity. Increased membrane fluidity may interfere with the dynamic assembly of actin cytoskeletal components essential to cell motility and focal adhesions essential for tumor viability and metastasis.

*Patent pending 2008

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A57.
Abstract A65: Identification of inflammation-associated plasma markers as response indicators in human colorectal cancer patients consuming freeze-dried black raspberries
Authors: Mentor-Marcel Roycelynn , Bobe Gerd , Sardo Christine , Wang Li-Shu , Albert Paul , Stoner Gary , Colburn Nancy
Published: May 2012

A65

Background

Inflammatory cytokines have been reported as biomarkers of colorectal adenoma risk in humans, however, little is known of biomarkers of response to interventions that may be useful as preventive or therapeutic agents for colorectal cancer. Freeze-dried black raspberries given to colorectal cancer patients were previously shown to be efficacious in altering cellular parameters associated with colorectal tumor development (i.e. apoptosis, cell proliferation, angiogenesis). We hypothesized that berry-induced changes in these cellular responses would be associated with changes in plasma levels of inflammatory cytokines that may serve as measures of efficacy of a 2-4 week dietary intervention of berries in colorectal cancer patients.

Methods

26 patients who had not received prior therapy consumed a slurry of black raspberries (20g in 100 ml drinking water) 3 times a day for 2-4 weeks. Plasma was collected before and after berry treatment and was analyzed for 9 inflammation-associated proteins.

Results

2 of 9 inflammation-associated plasma proteins were increased (i.e. IL-1β, GM-CSF) with berry treatment. Whether these diet-induced plasma changes correlate with changes in apoptosis, cell proliferation, and angiogenesis is being determined.

Conclusion

In summary, IL-1β and GM-CSF emerged as plasma proteins that were increased in colorectal cancer patients upon berry treatment. Changes in a subset of inflammation-associated plasma proteins may serve as response indicators to berry treatment under efficacious conditions and thus may be useful as a short term assay to monitor response to berry-based interventions for treatment of colorectal cancer.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A65.
Abstract A71: Molecular trail for the action of a metal-based drug in HPV positive cervical cancer cells
Authors: Srinivas Priya , Nair Rakesh , Kurup Maliyeckal Ramakrishna Panicker , Kuriakose Mini
Published: May 2012

A71

Cervical cancer is one of the most prevalent female cancers in world. The treatment for cervical cancer includes chemotherapy. Most potential standard chemotherapeutic agents have side effects. A need-based approach for developing new anti cancer agents from metal compounds with fewer side effects will have enormous implications. Opportunities exist to exploit metal-based drugs in the discovery and development of pharmaceuticals against cancers. Vanadium is a transition metal widely distributed in the environment, which is also a dietary micronutrient. The antidiabetic and anticancer activities of the oxovanadium complexes are recently known. In this study, human cervical cancer cell line SiHa, which is HPV16 - positive has been employed to determine the anticarcinogenic property of two oxovanadium complexes, OVK 49 and OVK 89. Among these OVK 49 proved to be more effective than OVK 89. Both complexes inhibited the growth of SiHa cells in a concentration and time dependent manner. Treatment of cells with oxovanadium complexes caused loss of mitochondrial membrane potential (ΔΨm) and morphological changes characteristic of apoptosis, such as the nuclear condensation. Moreover oxovanadium complex induced apoptosis, which involved release of mitochondrial cytochrome c and activation of caspase 8 and 9. The molecular mechanism behind the apoptotic induction due to these oxovanadium complexes is well established in this study. The E6 oncoprotein of human papillomaviruses has the potential to funtionally antagonize p53. Here it has also been investigated whether this reflects the regulation of p53 expression in HPV positive cervical cells. It has also been found that p21 was efficiently induced and HPV E6, E7 p53 and GADD45 were decreased by OVK 49 and 89 in SiHa cells. These results suggests that inspite of the presence of E6 protein HPV 16 positive cervical cancer cell lines are capable of responding efficiently to DNA damage provoked by metal based drug treatment through a p53 dependent pathway.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A71.
Abstract A75: DNA repair genotypes associated with benign breast disease in women at high breast cancer risk
Authors: Jorgensen Timothy , Helzlsouer Kathy , Hoffman Sandra , Hoffman-Bolton Judith , Crum Rosa , Visvanathan Kala
Published: May 2012

A75

Background

Benign breast disease (BBD) is a risk factor for breast cancer and may have a heritable component. Little is known about genetic factors associated with BBD risk. Deficient DNA repair has been implicated in the etiology of breast cancer. Since DNA repair occurs early in the breast carcinogenesis pathway, it is logical that polymorphic forms of DNA repair genes may alter BBD risk. No prior studies have reported on the association between DNA repair variants and BBD.

Methods

We examined the association between a panel of DNA repair genotypes and BBD in a sub-cohort of women within CLUE II, a population based cohort study in Washington County, Maryland. The women were followed from 1989 to 2003 with biennial questionnaires from 1996 onwards eliciting information on many health outcomes, including physician diagnosis of BBD and breast biopsies. 3,212 women reporting no prior diagnosis of BBD at baseline were genotyped for non-synonymous SNPs in 8 DNA repair genes (ERCC2, ERCC4, ERCC5, RAD23B, XPC, XRCC1, XRCC2, and XRCC4) that had previously been implicated in cancer risk. BBD-free survival time was fitted to age-adjusted Cox proportional hazards model and analyzed by DNA repair genotype.

Results

Carriers of the variant alleles of XRCC1 codon 194 and ERCC4 codon 415 were significantly more likely to be diagnosed with BBD than non-carriers -- hazard ratios (HR) = 1.36, 95%CI 1.06-1.74 and 1.39, 95%CI 1.09-1.76, respectively. The association between the ERCC4 variant and BBD was even greater among women with a positive family history of breast cancer. In this subgroup, the variant ERCC4 allele was associated with a 2.7 fold increase in breast cancer risk (HR = 2.63, 95% CI 1.52-4.66, p interaction = 0.02).

Conclusion

Variant ERCC4 and XRCC1 genotypes are significantly associated with a diagnosis of BBD in this population, particularly in women with a family history of breast cancer. These same genes have also been implicated in the risk of breast cancer in other studies. Taken together, these studies support the notion that these two DNA repair genes may have a common role in the etiology of both BBD and breast cancer.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A75.
Abstract A78: Common single nucleotide polymorphisms in immunoregulatory genes and multiple myeloma risk among women in Connecticut
Authors: Lee Kyoung-Mu , Dalsu Baris , Zhang Yawei , Hosgood H. Dean , Yeager Meredith , Zahm Shelia , Wang Sophia , Purdue Mark , Chanock Stephen , Zheng Tongzhang , Qing Lan
Published: May 2012

A78

Background

Increased risk of multiple myeloma has been observed among those who have a family history, suggesting a genetic component in multiple myeloma development. Also, a number of studies have shown that changes in immuoregulatory role of T cells may contribute to the development of multiple myeloma.

Objective

In light of the relationship between immune system dysregulation and multiple myeloma risk, we investigated whether genetic variation in 153 immune function genes are associated with multiple myeloma susceptibility in a population-based case-control study conducted among White Connecticut women.

Methods

Tagging single-nucleotide polymorphisms (SNPs; N=876) were selected using a pairwise linkage-disequilibrium based algorithm. Odds ratios (ORs) and 95% confidence intervals (CIs) for SNP genotypes were estimated using unconditional logistic regression. Tests of association for gene regions were conducted using the minP test. We applied the false discovery rate (FDR) method to the minP test results as a means of controlling for multiple comparisons. Haplotype analyses among subjects were performed using the haplo.stat statistical package in the R program. Haplotype frequencies were estimated from genotype data using the expectation-maximization algorithm while at the same time excluding those with frequencies less than 1%, and were evaluated by the global score test.

Results

Six out of 75 gene regions were significantly (P<0.05) associated with multiple myeloma using the MinP test. However, only the PTMS-LAG3-CD4-GPR162-LEPREL gene region remained noteworthy after adjustment for multiple tests using the FDR method (minP=0.0018; noteworthy at a level of FDR control of 0.14). In this region, a total of six SNPs in two genes (CD4 and LAG3) were significantly associated with risk (Ptrend<0.05), with the strongest association observed for rs11064392 (ORAG/GG=2.53, 95% CI=1.59-4.02; Ptrend=0.0001). Haplotype analyses also supported an association with rs11064392.

Conclusion

Our findings suggest that genetic variation in CD4 may influence susceptibility to multiple myeloma.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A78.
Abstract A79: Hormonal factors and breast cancer risk in nulliparous women
Authors: Schonfeld Sara , Pfeiffer Ruth , Lacey James , Hartge Patricia , Doody Michele , Greenlee Robert , Park Yikyung , Schairer Catherine , Schatzkin Arthur , Sigurdson Alice , Visvanathan Kala
Published: May 2012

A79

Background

Nulliparous women are a growing population in the U.S. that is known to be at elevated risk for breast cancer. A term pregnancy is believed to exert a protective effect on the ductal structures in the breast through terminal differentiation. It is biologically plausible that limited ductal maturation in nulliparous women renders the breast more susceptible to the effects of hormonal factors.

Methods

We assessed the association between hormonal and other factors and breast cancer risk by parity among postmenopausal women. Data were combined from four prospective cohort studies [The Breast Cancer Detection Demonstration Project Follow-up Study, the NIH-AARP Diet and Health Study, The Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial, and The United States Radiologic Technologist Study]. Each dataset was analyzed using multivariable Cox Proportional Hazards models to estimate hazard ratios (HR) and 95% confidence intervals (CI) separately for nulliparous and parous women for known risk factors. Study-specific results were combined using a meta-analytic random effects model.

Results

A total of 1,611 breast cancers were reported among 32,636 nulliparous women and 8,284 breast cancers among 207,763 parous women. The meta-analytic HRs for the one of the strongest risk factors, current use of menopausal hormone therapy for ≥5 years, were 1.46 (95% CI: 1.10-1.82) and 1.50 (95% CI: 1.42-1.59) for nulliparous and parous women, respectively. Patterns of risk were similar for nulliparous and parous women for other factors including ages at menarche and last menstrual period, body mass index, family history of breast cancer, and benign breast disease. Some study-specific variability was detected.

Conclusions

Similar associations between hormonal risk factors and breast cancer were observed for nulliparous and parous women. However, given their elevated baseline risk, these risk factors are likely to contribute to a greater absolute risk of breast cancer among nulliparous women.

Citation Information: Cancer Prev Res 2008;1(7 Suppl):A79.