Idiopathic pulmonary fibrosis (IPF) is a disease with an unknown etiology mainly characterized by a progressive decline of lung function due to the scarring of the tissue deep in the lungs. The overall survival after diagnosis remains low between 3 and 5 years. IPF is a heterogeneous disease and much progress has been made in the past decade in understanding the disease mechanisms that contributed to the development of two new drugs, pirfenidone and nintedanib, which improved the therapeutic management of the disease. The understanding of the cofactors and comorbidities of IPF also contributed to improved management of the disease outcome. In the present review, we evaluate scientific evidence which indicates IPF as a risk factor for other diseases based on the complexity of molecular and cellular mechanisms involved in the disease development and of comorbidities. We conclude from the existing literature that while much progress has been made in understating the mechanisms involved in IPF development, further studies are still necessary to fully understand IPF pathogenesis which will contribute to the identification of novel therapeutic targets for IPF management as well as other diseases for which IPF is a major risk factor.

Kidney cancer ranks among the top 10 cancers in the United States. Although it affects both male and female populations, it is more common in males. The prevalence rate of renal cell carcinoma (RCC), which represents about 85% of kidney cancers, has been increasing gradually in many developed countries. Family history has been considered as one of the most relevant risk factors for kidney cancer, although most forms of an inherited predisposition for RCC only account for less than four percent. Lifestyle and other factors such as occupational exposure, high blood pressure, poor diet, and heavy cigarette smoking are highly associated with its incidence and mortality rates. In the United States, White populations have the lowest prevalence of RCC compared to other ethnic groups, while Black Americans suffer disproportionally from the adverse effects of RCC. Hence, this review article aims at identifying the major risk factors associated with RCC and highlighting the new therapeutic approaches for its control/prevention. To achieve this specific aim, articles in peer-reviewed journals with a primary focus on risk factors related to kidney cancer and on strategies to reduce RCC were identified. The review was systematically conducted by searching the databases of MEDLINE, PUBMED Central, and Google Scholar libraries for original articles. From the search, we found that the incidence and mortality rates of RCC are strongly associated with four main risk factors, including family history (genetics), lifestyle (poor diet, cigarette smoking, excess alcohol drinking), environment (community where people live), and occupation (place where people work). In addition, unequal access to improvement in RCC cancer treatment, limited access to screening and diagnosis, and limited access to kidney transplant significantly contribute to the difference observed in survival rate between African Americans and Caucasians. There is also scientific evidence suggesting that some physicians contribute to racial disparities when performing kidney transplant among minority populations. New therapeutic measures should be taken to prevent or reduce RCC, especially among African Americans, the most vulnerable population group.

As an alternative therapeutic treatment to reduce or eliminate the current side effects associated with advanced prostate cancer (PCa) chemotherapy, a multifunctional double-receptor-targeting iron oxide nanoparticles (IONPs) (luteinizing hormone-releasing hormone receptor [LHRH-R] peptide-and urokinase-type plasminogen activator receptor [uPAR] peptide-targeted iron oxide nanoparticles, LHRH-AE105-IONPs) drug delivery system was developed. Two tumor-targeting peptides guided this double-receptor-targeting nanoscale drug delivery system. These peptides targeted the LHRH-R and the uPAR on PCa cells. Dynamic light scattering showed an increase in the hydrodynamic size of the LHRH-AE105-IONPs in comparison to the non-targeted iron oxide nanoparticles (NT-IONPs). Surface analysis showed that there was a decrease in the zeta potential values for drug-loaded LHRH-AE105-IONPs compared to the NT-IONPs. Prussian blue staining demonstrated that the LHRH-AE105-IONPs were internalized efficiently by the human PCa cell line, PC-3. In vitro, magnetic resonance imaging (MRI) results confirmed the preferential binding and accumulation of LHRH-AE105-IONPs in PC-3 cells compared to normal prostate epithelial cells (RC77N/E). The results also showed that LHRH-AE105-IONPs significantly maintained T-2 MRI contrast effects and reduced T-2 values upon internalization by PC-3 cells. These paclitaxel-loaded double-receptor-targeting IONPs also showed an approximately twofold reduction in PC-3 cell viability compared to NT-IONPs.

Abstract Prostate Cancer (PCa) is the second leading cause of cancer-related deaths among men in the United States. Due to the harsh side effects of conventional chemo- and radiotherapies, targeted drug delivery is now gaining the focus of cancer researchers. The key for success of any newly developed targeted drug delivery systems is the novel design of targeting peptides which have high binding affinities to differentially over-expressed receptors on the surface of PCa cells. Two over-expressed such receptors, which can be targeted to the cell surface of PCa cells are: Gonadotropin Releasing Hormone Receptor (GnRH-R), and Prostate Specific Membrane Antigen (PSMA). The analysis done from molecular dynamics simulation data gave an important information about each interaction; such as RMSD, The number of contacts between receptor and peptide within a cutoff distance of 5 Å and the binding free energies, ΔGGBSA of the interactions. In all systems, the back bone RMSD deviation of the receptor was calculated using the last 10 ns of the sampling process and averaged over all trajectories. The RMSD data deviated between 1.22 - 2.51 Å. The free binding energies or ΔGGBSA was calculated from the last 10 ns of the sampling process. The QH1 system has the more negative ΔGGBSA -36.15 kcal mol-1 and the QH4 system has the least negative ΔGGBSA of -18.4 kcal mol-1 among the eight systems of GnRH-R. QH2 system exhibits a highly interactive hydrogen bond (bond distance is 2.65 Å) between 134th position of the receptor amino acid sequence GLU and 5th position of the qh2 peptide's TYR. The ΔGGBSA is for the qh2 system is – 34.92 kcal mol-1. In the PSMA and its targeting peptide interactions, T2IA has the least negative ΔGGBSA of -47.37 kcal mol-1. A salt bridge was observed between the 10th position of the T2IA peptide sequence ARG and the 328 position of the 3RBU receptor's GLU. Based on our molecular modeling studies, we have determined the best targeting peptides among all the newly designed peptides. The binding efficiency of the targeting peptides with the lowest binding energies will be further investigated surface plasmon resonance (SPR) in near future. Citation Format: Ahmad Salam, Vincent Hembrick, Jesse Jaynes, Timothy Turner, Mohamed Abdalla. Molecular modeling of novel peptide-receptor interaction for targeted drug delivery of prostate cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2170.

Abstract Purpose: Prostate Cancer (PCa) is the second leading cause of cancer-related deaths among men in the United States. Due to the harsh side effects of conventional chemo- and radio-therapies, targeted drug delivery is now gaining focus of cancer researchers. The key for success for any newly developed targeted drug delivery systems is the novel design of targeting peptides which have high binding affinities to differentially over-expressed receptors on the surface of PCa cells. Four over-expressed receptors which can be targeted on the cell surface of PCa cells are: (GnRH), (EGFR), (PSMA), and (uPAR). Thus, we propose to study the binding affinity of tweleve novel designed targeting peptides (three peptides per receptor) for these over-expressed receptors. Initially, we will study the binding affinity using a computerized molecular modeling program. We will corroborate our modeling data by determining the dissociation constants of the peptide-receptor interaction using sensitive experimental tools such as miscroscale thermophoresis. Methods: The newly designed peptides were generated in pdb (protein database) format and the known pdb files of the receptors were collected from protein data bank. Cluspro 2.0, developed by Boston University, is a protein docking server with a molecular modeling program that was used to calculate the binding energies of these peptides to their corresponding receptors. The data obtained was analyzed using Pymol. Results: The molecular docking studies showed that the GnRH tarteting peptide QH10, uPAR targeting peptide CV13, EGFR targeting peptide RL10, and PSMA targeting peptide TH12 have the lowest mean binding energies for their respective top three binding conformations. The calculatd binding energies for the aftermentioed peptides-receptors combination are: -1285 KCal/M, -1029 KCal/M, -1019 KCal/M and -1037 KCal/M, respectively. There results suggested a very high interaction for these newly designed peptides and their corroponding receptors as the typical calculated binding energy for an antigen-antibody interaction is -700 KCal/M. Conclusion: Based on our molecular docking studies, we have determined the best targeting peptides among all the newly desiged peptides. The binding efficiency of the targeting peptides with the lowest binding energies will be further investigated using microscale thermophoresis. This work is supported by grant funding from NIH/RCMI G12MD00758 (T. Turner) and NIH/NCI U54 CA118623 (T. Turner & M. Abdalla). Note: This abstract was not presented at the meeting. Citation Format: Mohamed O. Abdalla, Ahmad Bin Salam, Vincent Hembrick, Manikanthan Bhavaraju, Clayton Yates, Jesse Jaynes, Timothy Turner. Molecular modeling studies of novel receptor targeted peptides in the treatment of prostate 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 3694. doi:10.1158/1538-7445.AM2015-3694

Abstract Purpose: As an alternative to the drawbacks of current advanced cancer treatments such as conventional chemotherapy, we propose a multifunctional double targeting drug delivery system that utilizes the combination of cancer-targeting peptides fused to amphiphilic polymer coated iron oxide nanoparticles (IONPs) and loaded with suitable anticancer drugs as the payload. Methods: PC3 human prostate cancer cell line will be used initially to test the efficiency of the proposed drug delivery system. Our target sites of choice are the luteinizing hormone releasing hormone receptor (LHRH-R) and the urokinase-type plasminogen activator receptor (uPAR), and docetaxel was selected as the payload. A modified LHRH (ligand for LHRH-R) and AE105 (ligand for uPAR) were conjugated to polymer coated IONPs according to the manufacturer's protocol. Results: Conjugated IONPs were characterized by gel electrophoresis and Dynamic Light Scattering (DLS). IONPs showed expected narrow size distribution after conjugation with peptide ligand. The average hydrodynamic size of non targeted IONPs (36.84 nm) was increased upon conjugation of double peptide to their surface (44.06 nm). Conjugation of peptides to carboxylic groups of polymer coating on IONPs resulted in a decrease of zeta potential from -70.43 mV to -58.06 mV. Prussian blue staining demonstrated that LHRH and AE105 conjugated IONPs were internalized efficiently by the PC3 cells. The drug loading and release capability of conjugated IONPs will be evaluated by HPLC. In addition the internalization of conjugated IONPs will also be examined utilizing Magnetic Resonance Imaging (MRI), and tumor eradication will be determined by MTT assay. Conclusions: We expect a significant enhancement in the binding efficiency of the LHRH-IONPs-AE105 to the prostate cancer cells, increased efficiency in tumor eradication, and better imaging and monitoring capabilities because IONPs can be visualized by MRI during molecular imaging of the prostate cancer cells. Therefore, we believe the optimization of the proposed system will enhance targeted nanomedicine and significantly improve the health outcomes and quality of life for cancer patients because of the following reasons: 1) its ability to deliver anticancer drugs specifically to cancer cells while sparing the surrounding normal tissues, and 2) the capability of in situ monitoring of the therapeutics. Citation Format: Md Shakir U. Ahmed, Mohamed O. Abdalla, Timothy Turner. Double targeting nanoscale drug delivery system for treatment and imaging of metastatic solid cancers. [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 4480. doi:10.1158/1538-7445.AM2014-4480

Here, we report improved solubility and enhanced colonic delivery of reduced bromonoscapine (Red-Br-Nos), a cyclic ether brominated analogue of noscapine, upon encapsulation of its cyclodextrin (CD) complexes in bioresponsive guar gum microspheres (GGM). Phase-solubility analysis suggested that Red-Br-Nos complexed with β-CD and methyl-β-CD in a 1:1 stoichiometry, with a stability constant (Kc) of 2.29 × 10(3) M(-1) and 4.27 × 10(3) M(-1). Fourier transforms infrared spectroscopy indicated entrance of an O-CH₂ or OCH₃-C₆H₄-OCH₃ moiety of Red-Br-Nos in the β-CD or methyl-β-CD cavity. Furthermore, the cage complex of Red-Br-Nos with β-CD and methyl-β-CD was validated by several spectral techniques. Rotating frame Overhauser enhancement spectroscopy revealed that the Ha proton of the OCH₃-C₆H₄-OCH₃ moiety was closer to the H₅ proton of β-CD and the H₃ proton of the methyl-β-CD cavity. The solubility of Red-Br-Nos in phosphate buffer saline (PBS, pH ∼ 7.4) was improved by ∼10.7-fold and ∼21.2-fold when mixed with β-CD and methyl-β-CD, respectively. This increase in solubility led to a favorable decline in the IC₅₀ by ∼2-fold and ∼3-fold for Red-Br-Nos-β-CD-GGM and Red-Br-Nos-methyl-β-CD-GGM formulations respectively, compared to free Red-Br-Nos-β-CD and Red-Br-Nos-methyl-β-CD in human colon HT-29 cells. GGM-bearing drug complex formulations were found to be highly cytotoxic to the HT-29 cell line and further effective with simultaneous continuous release of Red-Br-Nos from microspheres. This is the first study to showing the preparation of drug-complex loaded GGMS for colon delivery of Red-Br-Nos that warrants preclinical assessment for the effective management of colon cancer.

miRNA expression in African American compared to Caucasian PCa patients has not been widely explored. Herein, we probed the miRNA expression profile of novel AA and CA derived prostate cancer cell lines. We found a unique miRNA signature associated with AA cell lines, independent of tumor status. Evaluation of the most differentially expressed miRNAs showed that miR-132, miR-367b, miR-410, and miR-152 were decreased in more aggressive cells, and this was reversed after treatment of the cells with 5-aza-2'-deoxycytidine. Sequencing of the miR-152 promoter confirmed that it was highly methylated. Ectopic expression of miR-152 resulted in decreased growth, migration, and invasion. Informatics analysis of a large patient cohort showed that decreased miR-152 expression correlated with increased metastasis and a decrease in biochemical recurrence free survival. Analysis of 39 prostate cancer tissues with matched controls (20 AA and 19 CA), showed that 50% of AA patients had statistically significant lower miR-152 expression compared to only 35% of CA patients. Ectopic expression of miR-152 in LNCaP, PC-3, and MDA-PCa-2b cells down-regulated DNA (cytosine-5)-methyltransferase 1 (DNMT1) through direct binding in the DNMT1 3'UTR. There appeared to be a reciprocal regulatory relationship of miR-152/DNMT1 expression, as cells treated with siRNA DNMT1 caused miR-152 to be re-expressed in all cell lines. In summary, these results demonstrate that epigenetic regulation of miR-152/DNMT1 may play an important role in multiple events that contribute to the aggressiveness of PCa tumors, with an emphasis on AA PCa patients .

Abstract Prostate cancer is the most commonly diagnosed malignancy in men, with African American men experiencing a rate 60% higher than white patients. At the time of diagnosis, approximately 50% of men have clinically advanced disease. African American men have almost twice the incidence and death rates related to prostate cancer compared to Caucasian men. Several studies have suggested that some of these differences may be attributed to the elevated expression of different genes. Many factors have been associated with why prostate tumors in AA patients are more aggressive. However there is a lack of knowledge, whether this is the result of genes that promote aggressiveness or promote tumor development. Methylation profiling of prostate tumors has demonstrated that the loss of a number of key regulatory genes are not via mutation, but rather hypermethylation. This is particularly evident in the AA patients where hypermethylation of a number of genes in normal or pre-malignant areas are thought to predispose a full-blown malignancy. However the underlining mechanism of these acquired methylation patterns is poorly understood. To better understand the epigenetic regulation of genes associated with promoting aggressiveness and specifically to determine if this can explain the more aggressive nature of AA tumors , we conducted 662 microRNA microarray profiling utilizing novel isogenic non-malignant and malignant cell lines derived from both AA and CA tumor patients. Utilizing multiple criteria, included pathological stage and race, we were able to determine a distinct microRNA signature that was unique the AA derived cell lines, independent of tumor status. Validation by q-PCR of the 10 most significantly differentially expressed miRNA's across our 11 cell line panel demonstrated that loss of miR-152 expression was associated with aggressiveness. Restoring miR-152 expression in miR-152 deficient cell lines, resulted in decreased proliferation, and S phase arrest of the cell cycle. In silico informatics analysis predicted that miR-152 contains a significant number of CpG islands within the promoter region upstream of the start site. We confirmed this analysis with bisulfite conversion and sequencing of the promoter, as well as treatment with demethylation agent 5-Aza-2′-deoxycytidine. Furthermore, we observed an inverse relationship of miR-152 with predicted target DNA methyltransferase-1 (DNMT1), suggesting a reciprocal maintenance of hypermethylation for not only miR-152, but also a number of other methylated genes. This is plausible given the well-established role of DNMT1 in methylation of a large number of genes found to promote aggressiveness in prostate tumors . Lastly, a comparison of normal/tumor ratios of miR-152 expression in 20 CA and 20 AA prostate cancer patients, we observed significantly decreased expression in tumors from CA patients as expected, however this was not observed in AA patients. AA patients also displayed an overall lower expression compared to CA tumors. In summary, these results are the first to identify unique miRNAs that contribute to aggressive prostate cancers in AA patients. Furthermore epigenetic regulation of the miR-152/DNMT1 regulatory loop may play an import ant role in multiple events that contribute to the aggressiveness of PCa tumors. Citation Format: Shaniece C. Theodore, Honghe Wang, Jhong Rhim, Timothy Turner, Melissa Davis, William Grizzle, Clayton C. Yates. Functional biomarkers that promote African American prostate cancer through epigenetic regulation. [abstract]. In: Proceedings of the Sixth AACR Conference: The Science of Cancer Health Disparities; Dec 6–9, 2013; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2014;23(11 Suppl):Abstract nr PL01-02. doi:10.1158/1538-7755.DISP13-PL01-02

: Prostate cancer disproportionately afflicts African-American men. As such, we feel that it is critically important to recruit researchers from this population if we are to conquer this disease. Numerous programs have attempted to recruit minorities to biomedical research and prostate cancer in particular. Often this involves a short period of research immersion during a summer semester. However, it has been shown that many of these trainees do not persevere in the selected area due to the singular nature of the experience. Our goal is to formalize a program to broaden the scope of and enlarge Tuskegee University s prostate cancer research , which will be accomplished through tatargeting interested undergraduate students early during their science studies at Tuskegee University and enabling them to participate in summer research and education training periods at the University of Pittsburgh and the University of Pittsburgh Cancer Institute as part of their overall prostate cancer education.

Abstract Metastatic spread of prostate cancer (PCa) is responsible for the majority of prostate cancer-related deaths. Whereas socioeconomic factors, different geographic areas and ethnicity contribute to the incidence and mortality rates of this disease, they cannot fully explain the tumor biology. Our understanding of the mechanisms that drive PCa progression and metastasis is still limited. Kaiso belongs to a BTB/POZ zinc finger protein family and is known as a transcriptional repressor by binding to sequence-specific Kaiso binding sites or methyl-CpG dinucleotide pairs. Kaiso expression and localization have been reported to correlate with the prognosis and metastatic potential in several human malignancies. Previous studies from our lab showed that the progression of PCa is closely associated with the over-expression and nuclear translocation of Kaiso. Our objective here is to explore the potential molecular mechanisms underlying Kaiso mediated increase in PCa progression. MicroRNAs play an emerging role in cancer initiation and progression. Therefore we proposed to investigate Kaiso targeting microRNAs and their roles in PCa. PC-3 cells, a highly metastatic PCa cell line, were stably transfected with a shRNA plasmid against human Kaiso or a scramble control. Two clones exhibited >80% downregulation of Kaiso in PC-3 cells at either the transcriptional or translational levels were used for further studies. We determined miRNA expression profiles correlated with Kaiso knock down and DNMT1 inhibitor 5-Aza de-methylation treatment using microRNA arrays. Comparative analysis revealed 11 miRNAs were significantly altered by Kaiso knock down and 32 miRNAs were significantly altered by 5-Aza treatment (> 2 fold, p<0.05). Real-time PCR confirmed that miR-31 expression was up-regulated in both Kaiso knock down cells and 5-Aza treated cells. From the analysis of differentially expressed miRNAs, miR-31 expression was also negatively correlated with kaiso expression in a set of prostate cell lines. ChIP assay revealed that kaiso directly binds to miR-31 promoter. The restoration of miR-31 suppressed cell proliferation; induced apoptosis and inhibited tumor cell invasion and metastasis in vitro. Inhibition of miR-31 in PC-3 shKaiso cells restored kaiso knock down-inhibited cell proliferation; migration and metastasis. Xenograft model indicated that miR-31 overexpression significant suppressed tumor formation in vivo. Taken together, our studies suggest that Kaiso regulates specific miRNA expression in PCa cells, indicating that Kaiso could promote cancer progression through regulating miR-31 expression in a methylation dependent manner. Citation Format: Honghe Wang, ShaNekkia Black, Wei Liu, Fu Zhao, Timothy Turner, Clayton Yates. Transcriptional repressor Kaiso promotes cancer progression by targeting miR-31 in prostate cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3062. doi:10.1158/1538-7445.AM2013-3062 Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.

Abstract PURPOSE: African American men have almost twice the incidence and death rates related to prostate cancer compared to Caucasian men. Several studies have suggested that some of these differences may be attributed to the elevated expression of different genes. Recently, a new class of genetic regulators has emerged known as microRNAs. Many reports have found microRNAs to be implicated in cell growth, cell differentiation and the onset of many diseases including prostate cancer. We sought to investigate whether microRNAs are differentially expressed in African American and Caucasian prostate samples. METHODS: We analyzed the expression of over 300 microRNAs in 11 African American and Caucasian cell lines utilizing Affymetrix Microarray Chip. Validations of the most significant miRNAs were done by qRT-PCR. Methylation analysis of miR-152 promoter region was carried out by performing bisulfite modification and DNA sequencing. Characterization of the miRNA was carried out by performing MTT assay, Flow Cytometry, Western Blot, and Imunoflourescence. RESULTS: We identified 15 microRNAs that were differentially expressed between the African American and Caucasian cell lines. Further validations led to the identification of miR-152 which is significantly expressed among both groups. Bisulfite modification and DNA sequencing revealed that miR-152 is methylated in aggressive prostate cancer cell lines. 5-Aza-2’/TSA treatment restored miR-152 levels in these cell lines. MiR-152 is responsible for inhibiting DNA methyltransferase 1, Rictor and TGF-β. Our results showed that miR-152 reduced cellular proliferation in PC-3, DU-145 and LNCaP prostate cancer cell lines. In addition, miR-152 initiated G2-M cell cycle arrest. Expression of miR-152 in 40 normal-tumor paired prostate tissue samples indicated that its expression was down-regulated in 67% of tumor samples. Analysis of African American and Caucasian prostate normal-tumor paired sample revealed that both normal and tumor prostate samples had lower miR-152 expression in the African American prostate samples compared to the Caucasian prostate samples. CONCLUSION: These results give evidence that microRNAs may play a role in the advance progression seen in prostate cancer in African Americans. Future studies will focus on validating these results in larger patient sample sizes. Research supported by G12 RR03059-21A1(NIH/RCMI), and CA118623 (NIH/NCI). Citation Format: Shaniece Theodore, Melissa Davis, Jhong Rhim, William Grizzle, Timothy Turner, Clayton Yates. MicroRNA expression profiling in African American and Caucasian prostate cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3060. doi:10.1158/1538-7445.AM2013-3060

Lytic peptides represent a novel class of therapeutics that from our view, have not received much attention clinically for the treatment of common cancers. This is partly due to the lack of tumor specificity of these compounds compared with other classes of therapies which enhances the possibilility of unwanted side effects. Herein we detail the use of lytic peptides from their initial use as anti-bacterical/anti-fungal agents to now prospective cancer therapeutic agents. Additionally, we describe a novel design approach that facilitates the modulation of physical peptide characteristics and how this translates lytic activity.

Abstract Quercetin, one of the most abundant polyphenols, has been evaluated for its potential cancer preventive functions and for its anticancer activity in vitro and in vivo. Its interactions with chemotherapeutics, however, have not been established. Some reports indicate enhancement of drug activity by quercetin but others suggest precautions in co-administering antioxidants such as quercetin and chemotherapeutic drugs. Therefore, further investigations were needed to determine the conditions under which the co-treatment of cancer cells with drugs and quercetin could be clinically beneficial. This study was done to examine how quercetin modulates responses to the chemotherapeutic drugs, 5FU, camptothecin, and VP16, in colon and prostate cancer cells, and to determine the biochemical processes involved in the interactions. Survival of HCT116 colorectal cancer cells (p53 wild-type & null) was determined by assay of clonogenicity of cells exposed to selected concentrations of 5-FU, camptothecin, or VP16 in the presence or absence of quercetin. For cell cycle and biochemical experiments, cells were exposed for 24 hours to 10 µM 5FU, 1 µM camptothecin, or 10 µM VP-16 with or without 50 µM quercetin. The effects of the drugs on the cell cycle were measured by flow cytometry. The induction of p53 and its transcriptional targets, p21 and Bax, as well as the levels of cell cycle regulators, cyclin B1 and survivin, were determined by immunoblotting. Additionally, cell migration assays were used to evaluate the effect of combination treatments on the motility of RKO colorectal and PPC1 prostate cancer cells. Quercetin synergistically inhibited the clonogenicity of the wild-type HCT116 cells, but also inhibited effects on the cell cycle of all the drugs tested. In contrast, for p53-null cells, the combination of low concentrations of 5-FU with up to 6 µM quercetin promoted clonogenic survival. Exposure of wild-type cells to 50 µM quercetin reduced drug-induced up-regulation of p53, p21, and Bax. Combinations of quercetin and the drugs also reduced the levels of cyclin B1 and survivin. RKO and PPC1 cells exposed to the drugs had reduced migratory capacity. Although quercetin alone reduced their migratory capacity, combination treatments did not further reduce cell migration. In summary, quercetin in combination with 5-FU reduced the survival of p53-proficient HCT116 colorectal cancer cells independently of p21 and Bax, but combination of this flavonoid with low concentrations of 5-FU provided a survival advantage for p53-null cells. Further investigations are needed to determine the mechanisms and circumstances under which the combinations of bioactive dietary compounds and chemotherapeutic drugs are beneficial. Supported by grants from NCI/NIH (2U54-CA118948-06, and SC2CA138178). Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4664. doi:1538-7445.AM2012-4664

Advanced stages of cancer are characterized by increased aggressiveness, invasiveness, and the downregulation of tumor suppressor genes via methylation. Kaiso, a bi‐modal transcription factor, interacts with DNA through either a DNA consensus sequence or methylated CpG di nucleotides thus regulating gene expression. A clinical role for Kaiso expression in advanced stages of breast cancer remains unclear. Here we investigate the ability of kaiso to decrease the metastatic functional characteristics of breast cancer progression. Immunohistochemistry was performed to examine protein expression and localization in human breast tissue. siRNA kaiso treated cells were used to assess cell motility and invasion. An overall increase in Kaiso expression was found in primary tumor samples and this increased as cancer progressed to distant sites. Additionally, nuclear localization of Kaiso was observed in primary tumor and lymph node metastasis in breast tumors. Construct treated cell lines showed a delay in cell motility, invasion. 5‐aza‐2′‐deoxycytidine treated cells showed a re‐expression of tumor suppressor E‐cadherin, which correlated with E‐cadherin re‐expression in siRNA Kaiso treated cells. Downregulation of E‐cadherin, migration and invasion are key to cancer progression and metastasis. Our results reveal kaiso as a potential indicator and/or therapeutic agent of breast cancer metastasis.