bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2219: Inhibitor of differentiation-1 is a novel prognostic factor among NSCLC patients with adenocarcinoma histology and contributes to therapy resistance
Authors: Ponz-Sarvise Mariano , Nguewa Paul A. , Pajares Maria J. , Agorreta Jackeline , Lozano Maria D. , Redrado Miriam , Pio Ruben , Behrens Carmen , Wistuba Ignacio , Garcia-Foncillas Jesus , Montuenga Luis M. , Calvo Alfonso , Gil-Bazo Ignacio
Published: June 2012

High Id1 levels have been found in some tumor types, particularly in advance stages. We aimed to study Id1 levels and their prognostic value in a large series of different histologies of stage I-IV NSCLC patients and to test Id1 function in cell lines and cells derived from malignant pleural effusions (MPE) from patients (pts). Id1 expression was analyzed in NSCLC samples from 311 pts and 65 normal lung tissues by immunohistochemistry. Id1 mRNA expression was also studied in 111 pts using publicly available microarrays. Significantly higher Id1 protein expression levels were found in tumors compared to normal tissue (p<0.001) and in adenocarcinomas (AC) compared so squamous histology (p<0.001). Among the localized NSCLC pts undergoing surgery, higher Id1 expression levels were associated with a shorter overall survival (OS) for AC histology (p=0.017). In the surgery+adjuvant chemo-radiotherapy pts, the same correlation was found between Id1 levels and OS among AC samples (p=0.038). Consistently, in stage IV pts receiving palliative chemotherapy, Id1 maintained its prognostic value (shorter OS in pts with higher levels) for AC (p=0.003). Id1 was also correlated with time-to-progression (TTP) after chemotherapy (lower levels, prolonged TTP) in AC pts (p=0.008). The in silico analysis confirmed this association. High expression of Id1 was also found by western blots in squamous cell lines (H520, HCC15, H157 and H58) and in lesser extent in AC cell lines (SKLU-1, LXF-289, H322, H23) and MPE-derived AC cells. Id1 knockdown resulted in a significant reduction of the clonogenic activity of squamous and AC cells. Id1 silencing on radiotherapy and chemotherapy-resistant MPE-derived cells restored sensitivity to both therapies. We conclude that Id1 protein and mRNA expression is an independent prognostic factor among pts with AC but no other histologies, regardless of the stage or treatment received. In cell lines derived from MPE pts, Id1 downregulation decreases NSCLC cell proliferation and sensitizes cells to radiotherapy and chemotherapy, in vitro.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2219. doi:10.1158/1538-7445.AM2011-2219
Abstract 2222: Identify and evaluate novel biomarker CIP2A for cervical cancer diagnostics
Authors: Huang Lisa P. , Libby Erika , Trama Jason
Published: June 2012

Early detection of cervical cancer is critical for a favorable prognosis. HPV infection is not a reliable marker for predicting the malignancy potential of cervical lesions. Standard cytological detection methods such as Pap smear are highly subjective. As a result, there remains a very high demand for a diagnostic assay capable of determining which early stage cervical lesions will progress to cervical cancer. We sought to identify cervical cancer biomarkers and to test the hypothesis that the small percentage of HPV positive patients who progress to cervical cancer have aberrant expression of the cell signaling component, c-myc and its regulators, including protein phosphatase 2A (PP2A) and cancerous inhibitor of PP2A (CIP2A).

In this study, Bio-Plex liquid microarray analysis was used to determine the HPV status cervical specimens (NNon-tumor=28, NTumor=30). qRT-PCR, Western blotting, flow cytometry, and ELISA analysis were used to access expression levels of CIP2A, c-myc, PP2A, Ki67, TOP2A, MCM2, MCM5, p14ARF, and p16INK4a.

Our studies indicate CIP2A was abundantly expressed in cervical cancer cell lines and it was not expressed in normal epithelial cells. CIP2A mRNA levels were higher in cervical tissues from stage I, II, and III cervical cancer patients in comparison to the level of CIP2A mRNA in normal tissue. CIP2A protein was specifically expressed in cervical cancer tissues at different cancer stages, and not in non-cancer or adjacent non-tumor cervical tissue. Our data showed that CIP2A mRNA detection had a sensitivity of 80% and specificity of 91% and CIP2A protein expression detection had a sensitivity of 80% and specificity of 100%, similar to that of p16INK4a biomarker. Although CIP2A/PP2A/c-myc pathway may work together in the development of cervical cancer, our data strongly indicate that only CIP2A (but not PP2A or c-myc) is a reliable biomarker for detection of cervical cancer. In this study, we also examined the combined use of at least two biomarkers as a tool to detect cervical cancer. Our studies also suggest there was no strong correlation of CIP2A expression with HPV subtype, age, ethnical background, or other patient characteristics.

In conclusion, our clinical studies suggest that CIP2A is a reliable and sensitive biomarker for detection of cervical cancer. CIP2A mRNA or protein can both serve as reliable markers. A combined use of CIP2A with at least on additional marker selected from the group consisting of Ki67, TOP2A, MCM2, MCM5, and p16 will improve the assay sensitivity and specificity. CIP2A is a novel biomarker for early detection of cervical cancer in humans.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2222. doi:10.1158/1538-7445.AM2011-2222
Abstract 2226: Intraoperative harvesting of human fallopian tubes for cytodiagnostic and pathobiological studies
Authors: Esposito Nicole N. , Kruk Patricia A. , Hoffman Mitchell , Glazerman Larry R. , Nicosia Santo V.
Published: June 2012

Background: Fallopian tube intraepithelial cancer is a potential precursor of high-grade, sporadic and familial, pelvic carcinomas. Materials and Methods: Using a protocol for rapid intraoperative cytology (IC), fallopian tube fimbrial epithelium (FTFE) was harvested from 24 women (age: 40-80) undergoing surgery for non-neoplastic (N = 14, leiomyomas, adenomyosis, ovarian and paraovarian serous cysts, endometriosis) and neoplastic (N = 10, serous and endometrioid carcinomas with tubal involvement) lesions. Under IRB-approved protocol, eight cytological imprints were obtained from each fimbria and stained intraoperatively by Diff-Quik® and Papanicolaou for general morphology, by Trypan blue to assess cell viability and by rapid immunohistochemistry for evaluation of cytokeratin (CAM 5.2), Ki67 (MIB-1) p53 and high motility group protein (HMGA2) expression. Results: Tubal imprints yielded 0.5-1 x106, cytokeratin-positive and > 85% viable cells/patient with a 2:1 distribution of ciliated: mucous cells dispersed within a proteinaceous material of variable density with rare microcalcifications; reactive fibroblasts and mesothelial cells represented < 3% of cell harvests with the highest percentage seen in endometriosis and malignancies. Malignant fimbrial imprints (8 serous and 2 endometrioid carcinomas) exhibited diffuse accumulation of p53 and HMGA2, a Ki67 proliferation index of 50-70% and displayed diagnostic malignant criteria in all cases. Conclusions: Tubal harvest and intraoperative cytology of fallopian tubes allows for: a) rapid evaluation of tubal cancer, currently viewed as the main source of high-grade serous ovarian carcinomas; and b) triaging of cellular material for basic and translational studies including molecular cancer pathobiology and biomarker discovery. The technique will also provide a useful diagnostic adjunct to emerging in vivo high-resolution optical technologies. (Supported by Ovarian Cancer Research Fund).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2226. doi:10.1158/1538-7445.AM2011-2226
Abstract 2229: Detection of somatic mutations in KRAS gene in Chilean patients with advanced colorectal cancer
Authors: Hurtado Claudia , Zárate Alejandro J. , Kronberg Udo , Lévican Jorge , Contreras Luis , López-Köstner Francisco
Published: June 2012

Introduction: At present, the application of tailored oncologic therapies requires previous performance of mutational analysis of certain genes. In patients with advanced colorectal cancer (CRC), it is essential to detect the presence of activating mutations in the KRAS gene to predict the effectiveness of certain treatments.

Purpose: To determine the presence of KRAS mutations in Chilean patients with advanced CRC, and correlate the mutational status with clinical-pathological data.

Methods: Between October 2009 and August 2010, we prospectively studied the presence of somatic mutations in the KRAS gene in codons 12 and 13 in biopsies of patients with advanced colorectal cancer by allele specific and real time PCR kit. The results were registered in a database, along with the pathology results and clinical data of the patients. Statistical analysis was performed by Student T test and Chi-square test. A p>0.05 was no significant.

Results: A total of 123 patients were analyzed, with a mean age of 61 years and male predominance (60%). Mutations were detected in 47 (39%) of them and only in 2 cases the study was inconclusive. The distribution of the different mutations was as follows: 12Asp (32%), 12Val (26%), 13Asp (19%), 12Cys (11%), 12Ala (6%) and 12Ser (6%). Separated by gender, the detection rate was higher in male patients (45% vs 30%) (p=0.59). Analyzing the data by location of tumor, we found that mutations in right colon tumors was present in 26% (6/23), whereas tumors of the left colon and sigmoid were 43% (16/37) (p=0.1). Mutations detected in patients with lymph node metastasis was 42% comparing to node-negative patients (32%) (p=0.3). In patients with poorly differentiated tumors we found 50% mutations compared to well differentiated (18%) (p=0.08). Finally, the mortality was similar when we compared between WT and mutations patients.

Conclusions: In Chile, 39% of patients with advanced CRC carry mutations in KRAS gene. This result is similar to that described in other populations. There is a tendency to a higher mutation rate in males, patients with left colon cancer, poorly differentiated tumors and patients with lymph nodal involvement.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2229. doi:10.1158/1538-7445.AM2011-2229
Abstract 2245: Fever-range whole-body thermochemotherapy (FR-WB-TT + chemo-biotherapy) can induce long-duration response in both metastatic small-cell lung cancer (SCLCA) and other chemotherapy resistant neuroendocrine cancers
Authors: Bull Joan Marie C , Dancsak Theresa , Su Young , Li Shuang , Redwine Michael D. , King Terri M. , Brown Robert E.
Published: June 2012

Introduction: Metastatic small-cell lung cancer (SCLCA) often relapses quickly after chemotherapy and survival thereafter is short. Resistant neuroendocrine cancers of other origins also have a poor prognosis. Methods: A schedule-optimized Phase I-II thermochemotherapy regimen combining FR-WB-TT with cisplatin (CIS), gemcitabine (GEM), and metronomic interferon-α (IFN) was tested. The treatment regimen used an explicitly sequenced combination of CIS (60 mg/m2) given day 1 with weekly IFN (100 mcg. peginterferon-α-2). On day 3, thirty-six hours later, FR-WB-TT (6h at 40°C) was administered with GEM (600 mg/m2). GM-CSF was administered daily for 10 days beginning 1 week after GEM. The cycle was repeated every 28 days. Results: We have treated 14 patients with metastatic neuroendocrine cancer; 3 with SCLCA and 11 with other neuroendocrine cancers (4 large-cell neuroendocrine of lung, 3 pancreas, 2 prostate, 1 H&N, & 1 adrenal). Thirteen (93%) received prior chemotherapy. The number of treatments ranged from 1-6 (median 4). Response: Using RECIST criteria, we documented: 2 complete responses [CR; 14 %; both SCLCA, durations 45+ months (m), 52 m], 6 partial responses (PR; 43%; >90% tumor response; mean duration 25 m, range 8-37 m; 2 prostate, 2 pancreas, 1 H&N, 1 adrenal). Three patients had stable disease (SD; 21%; all > 4 m duration; 2 large-cell of lung, 1 pancreas). Three patients had progressive disease (PD; 21%; 2 large-cell neuroendocrine of lung, 1 SCLCA). In summary, we observed a 57% overall response (CR + PR). Toxicity: We documented grade 2 leucopenia in twelve patients (86%), grade 2 thrombocytopenia in eight patients (57%), and grade 1 thrombocytopenia in six patients (43%). All patients developed grade 2 anemia. One patient (7%) had a small, grade 2, skin burn. We observed no neuropathy or ototoxicity. Although not formally documented, responding patients experienced increased quality of life with greatly decreased/elimination of pain, weight gain, and resumption of normal activity. Conclusion: This multimodality FR-WB-TT regimen appears to be well tolerated, and demonstrates good clinical activity of relatively long duration in metastatic SCLCA and other resistant stage IV neuroendocrine cancers.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2245. doi:10.1158/1538-7445.AM2011-2245
Abstract 2252: ALDH7A1 expression is associated with recurrence in patients with resected non-small cell lung carcinoma
Authors: Giacalone Nicholas J. , Eisenberg Rosana , Chen Heidi , Olsen Sandra , Massion Pierre , Carbone David P. , Dang Thao P. , Lu Bo
Published: June 2012

The aldehyde dehydrogenase (ALDH) enzymes are important for detoxification of endogenous and exogenous aldehydes. Expression of ALDH family members has recently been described as a potential marker for tumor-initiating cancer stem cells in a variety of human malignancies, including lung cancer. We were interested to determine whether expression of ALDH7A1, a member of the ALDH family, has prognostic significance in resected non-small cell lung carcinoma (NSCLC). Tumor specimens were obtained for 107 patients with completely resected stage I through stage III NSCLC from paraffin-embedded tissue microarrays and stained with an antibody specific for ALDH7A1. Staining patterns were graded by a pathologist based on the intensity of staining and the percentage of cells stained. A staining score index was calculated by multiplying intensity score by the percentage area with positive staining. An index of ≥100 was considered positive, while an index <100 was deemed negative, as determined by visually determining a clear positive stain supported by histograms of the range of scores. Staining index was subsequently correlated with clinical data. Positive ALDH7A1 staining was identified in 46 patients, and negative staining was noted in 48 patients, with 13 tumor sections missing. Multivariate analysis demonstrated that NSCLC patients with positive ALDH7A1 expression had decreased overall survival relative to ALDH7A1-negative tumors, although this did not reach significance (hazard ratio 1.42, 95% confidence interval 0.84 to 2.389; p=0.192). However, patients with ALDH7A1-expressing NSCLC tumors had a significantly higher incidence of lung cancer recurrence compared to patients with ALDH7A1-negative tumors (hazard ratio 6.24, 95% confidence interval 2.06 to 18.86; p=0.001). These data indicate that ALDH7A1 staining is present in a substantial number of NSCLC tumors and may be a biomarker predictive for increased incidence of cancer recurrence in patients with surgically resected NSCLC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2252. doi:10.1158/1538-7445.AM2011-2252
Abstract 2255: Diagnostic and prognostic value of oncogenic human papillomavirus in patients with carcinoma of unknown primary of the neck
Authors: Speel Ernst-Jan M. , Straetmans Jos M.J.A.A. , Vent Julia , Mujagic Zlatan , Henfling Mieke , Haesevoets Annick , Haidl B , Huebbers Christian U. , Ramaekers Frans CS , Kremer Bernd , Klussmann Jens P.
Published: June 2012

Objective. To date, there is no consensus on the optimal diagnostic and therapeutic strategy for patients with carcinoma of unknown primary (CUP) of the neck. These tumors are heterogeneous in their clinical and biological characteristics, and a pre-operative prognostic marker is desirable to optimize therapy and improve outcome and survival. Human papillomavirus (HPV) is an etiologic factor in a subgroup of head and neck squamous cell carcinomas and has been identified as a significant prognostic biomarker. We sought to determine if HPV also may add relevant information on the origin and prognosis of these tumors.

Material and method. 47 patients (mean age 58.7 years; 40 men, 7 women) presenting with CUP of the neck between 1994 and 2008 in Cologne were examined by standard diagnostic procedures including radiological imaging of the head, neck, thorax and abdomen, positron emission tomography (PET), panendoscopy, curettage of the nasopharynx, bilateral tonsillectomy and blind probes of the base of tongue. All patients were surgically treated with neck dissection of the diseased neck as well as adjuvant chemoradiation. The mean follow up time was 34 months.

Formaldehyde-fixed, paraffin-embedded tissue specimens of the 47 metastases were examined retrospectively by means of p16INK4A immunohistochemistry, HPV-specific PCR and fluorescence in situ hybridization. Results were correlated with clinical follow-up data.

Results. Oncogenic HPV was present in 12/47 (26%) metastases (10 with HPV 16), which showed also p16INK4A overexpression. In 42% of CUP patients the primary tumor was discovered during follow-up. A significant correlation between HPV positivity and later detection of the primary tumor in the oropharynx was found (p=0.038). Moreover, patients with a p16INK4A- (13/47) or HPV-positive tumor had a more favorable overall 5-year survival rate then p16INK4A- and HPV-negative tumors (69% vs 33%, p=0.05; 65% vs 37%, p=0.093; respectively).

Conclusion. HPV is present in a quarter of neck metastases of CUP patients and the presence of oncogenic HPV and p16INK4A expression can serve to locate the primary tumor in the oropharynx. In addition, both biomarkers are indicators of a favorable prognosis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2255. doi:10.1158/1538-7445.AM2011-2255
Abstract 226: Transcriptional control of the BRCA1 expression by a metabolic switch
Authors: Di Lijun , Fernandez Alfonso , Gardner Kevin
Published: June 2012

Deletion or loss of BRCA1 function is the major cause of most familial forms of breast cancer, decreased expression of non-mutated BRCA1 alleles is often found in sporadic breast cancer, suggesting that alterations in BRCA1 transcriptional regulation may contribute to non-inherited forms of breast cancer. BRCA1 plays a central role in maintenance of genome stability and cell cycle control and therefore, often recognized as both a “care taker” and “gate keeper” of the genome. In this study, we show that BRCA1 expression is controlled by a tightly regulated dynamic balance between the association of transcriptional co-activators and co-repressors that regulate histone acetylation and chromatin accessibility at the BRCA1 promoter during the transcriptional response to estrogen stimulation. A central regulatory step in the control of the BRCA1 promoter is the eviction of the co-repressor and metabolic sensor C-terminal-binding protein1 (CtBP1). Loss of CtBP1 from the BRCA1 promoter either by estrogen induction, CtBP1 RNAi depletion or increasing NAD+/NADH ratio results in elevated histone acetylation, HDAC1 dismissal from the BRCA1 promoter and increased BRCA1 transcription. These findings indicate that BRCA1 expression is controlled by a metabolic switch that dynamically influences the localized deposition of epigenetic marks and chromatin structure at the BRCA1 promoter and suggests that caloric intake may influence the levels of this critical tumor suppressor in mammary tissues. Strikingly extended observation of other DNA damage response genes indicate that they are also under direct regulation by CtBP and CtBP complex. These data suggest a global molecular linkage between cellular metabolic status and the capacity to maintaining the genome free from damage or mistakes.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 226. doi:10.1158/1538-7445.AM2011-226
Abstract 2263: Prognostic significance of miR-215 in colon cancer
Authors: Karaayvaz Mihriban , Pal Timothy , Song Bo , Zhang Cecilia , Georgakopoulos Penelope , Mehmood Saira , Burke Stephanie , Shroyer Kenneth , Ju Jingfang
Published: June 2012

Background: We have previously shown that miR-215 suppressed the expression of key targets such as thymidylate synthase (TS), dihydrofolate reductase (DHFR), and denticleless protein homolog (DTL) in colon cancer. miR-215 is a tumor suppressor candidate due to the up-regulation of p53 and p21 by targeting DTL. However, high levels of miR-215 conferred chemoresistance due to cell cycle arrest and reduced cell proliferation by suppressing DTL. In this study, the clinical significance of miR-215 was further investigated as a potential prognostic biomarker in colon cancer patients.

Methods: Total RNAs were extracted from 34 paired normal and colon (stage II and III) tumor specimens using Trizol based approach. The levels of miR-215 and a closely related miR-192 were quantified using real time qRT-PCR expression analysis. The expression of DTL mRNA and protein were quantified by real time qRT-PCR and immunohistochemistry.

Results: The expression levels of miR-192 (p=0.0008) and miR-215 (p<0.0001) were significantly decreased in colon tumors compared to normal tissues. DTL was significantly over-expressed and was inversely correlated with miR-215, further suggesting an in vivo physiological relevance of miR-215 mediated DTL suppression. Kaplan-Meier survival analysis revealed that high levels of miR-215 expression (hazard ratio, 0.36; log rank test, p=0.04) are closely associated with poor patient's overall survival. Furthermore, an elevated expression of a miR-215 target protein DTL was detected in colon cancer tissues while no expression was present in normal tissues.

Conclusion: miR-215 has a unique potential as a prognostic and diagnostic biomarker in stage II and III colon cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2263. doi:10.1158/1538-7445.AM2011-2263
Abstract 2271: Single nucleotide polymorphisms in p53 codon 72 is associated with grade III tumors in African Americans with breast carcinoma
Authors: Shanmugam Chandrakumar , Salih Chura , Katkoori Venkat R. , Bovell Liselle L. , Callens Tom , Messiaen Ludwine M. , Hameed Omar , Grizzle William E. , Manne Upender
Published: June 2012

Background: Breast carcinomas (BCs) are more aggressive in African Americans (AAs) than in non-Hispanic Caucasian Americans (NHCAs); the reasons for this difference have not been fully elucidated. We recently demonstrated that single nucleotide polymorphisms (SNPs) at codon 72 of the p53 gene are significantly associated with a worse outcome in AAs with colorectal carcinomas. Therefore, we determined the prevalence of such SNPs in BCs and its possible association with adverse clinical or pathological features of BCs in AAs as well as NHCAs.

Methods: To determine the phenotypic status at codon 72, DNA samples, extracted from 137 BCs and their matching benign tissues (from 47 AAs and 90 NHCAs), were analyzed for genotypes by direct sequencing of exon 4. Also, these samples were evaluated for microsatellite (MS) instability by use of the ABI 3100 system. The results obtained were correlated with the demographic and clinicopathological features of the BCs.

Results: Overall, SNPs leading to either Arg/Pro or Pro/Pro at codon 72 were observed in 102 of 137 (74%) BCs, with a slightly higher prevalence in AAs compared to NHCAs (39 of 47; 83% vs. 63 of 90; 70%). The most frequent phenotype was Arg/Pro (84 of 137; 61%), followed by Arg/Arg (35 of 137; 26%) and Pro/Pro (18 of 137; 13%). The prevalence of the homozygous Pro/Pro phenotype, known to be associated with aggressive tumor behavior, was higher in AAs compared to NHCAs (19% vs. 10%). BCs with Pro/Pro or Arg/Pro SNPs were significantly more prevalent in grade III tumors (84%) than in grade I and II tumors (60%) (P=0.005); specifically, this distribution was highly significant in AAs (100% vs. 53%; χ2P=0.0001) as compared to NHCAs (72% vs. 62%; χ2P=0.451). Also, in AAs, BCs with the Pro/Pro phenotype were more frequently associated with MS stable status compared to the other phenotypes (63% vs. 18%; χ2P=0.011). Furthermore, Pro/Pro phenotype was associated with a shorter median survival in AAs (39.9 months) compared to NHCAs (112.2 months).

Conclusions: These preliminary findings suggest that SNPs at codon 72 of p53 gene are associated with aggressive tumor behavior in AAs. Additionally, the Pro/Pro phenotype is associated with MS stable disease and with a shorter median survival in AAs but not in NHCAs. These studies were supported by the UAB-SPORE grant (a pilot study and a supplement) from NCI/NIH (5P50CA089019) and by the Susan G. Komen Breast Cancer Foundation (POP0601383).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2271. doi:10.1158/1538-7445.AM2011-2271
Abstract 2274: The association of preoperative serum level of lipocalin2 with breast cancer prognosis
Authors: Sung Hyuna , Lee Sang-Ah , Lee Kyoung-Mu , Lee Ji-Young , Oh Jungmin , Lee Yunhee , Li Lian , Park Sue K. , Yoo Keun-Young , Noh Dong-Young , Ahn Sei-Hyun , Kang Daehee
Published: June 2012

Purpose: The association between the expression level of lipocalin2 with cancer progression has already been reported in several tumors such as lung, colon, and breast cancer. However, no previous study has examined the relationship between the circulating level of lipocalin2 and its effect on the prognosis of breast cancer. Thus, this study aimed at assessing whether the preoperative serum level of lipocalin2 is related to the risk of recurrence and death in patients who have undergone curative surgical treatment for breast cancer.

Design: A total of 370 histologically proven breast cancer patients who had undergone curative resections of the tumor between Mar 2004 and Jan 2007 were included in this study. The median follow-up time of survivors was 4.35 years. The preoperative serum level of lipocalin2 was assayed by using enzyme-linked immunosorbent assay. Disease-free survival was defined as the time from surgery to the date of the first locoregional recurrence, first distant metastasis, or death from any cause. Univariate survival analysis was performed using the Kaplan-Meier method, and log-rank tests were employed for comparison of survival curves. Multivariate analyses were conducted using Cox's proportional hazard regression model adjusted for age, tumor size, lymph node metastasis, hormone receptor status, adjuvant chemotherapy and hormone therapy.

Results: The Kaplan-Meier curve showed that patients with upper three-quarters of lipocalin2 concentration combined had lower survival rates than the patients with lowest quarter concentration (P=0.014). In multivariate analysis, lipocalin2 remained an independent prognostic marker for disease-free survival after adjusting for known prognostic factors. The hazard ratio comparing the uppermost quartile to the lowest quartile of lipocalin2 was 2.33 (95% confidence interval, 1.19-4.56; P=0.015).

Conclusions: The results revealed that patients with higher level of lipocalin2 showed significantly lower disease-free survival than patients with lower level. Our study suggests that the higher preoperative level of lipocalin2 may be closely linked to poor prognosis in breast cancer. Further validation is required and functional studies are warranted to elucidate the underlying biological mechanisms for the association.

Key words: Lipocalin2 (LCN), serum biomarker, breast cancer, prognosis

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2274. doi:10.1158/1538-7445.AM2011-2274
Abstract 2289: Potential role for Src kinase inhibitors in PKCα-overexpressing breast cancer
Authors: Molloy Mary E. , Tonetti Debra A.
Published: June 2012

The purpose of this study was to evaluate the role of Src kinase in estrogen induced growth inhibition in the T47D/PKCα breast cancer cell line. Our lab has previously shown that constitutive overexpression of protein kinase Cα (PKCα) imparts a tamoxifen-resistant, hormone-independent phenotype in the T47D:A18 breast cancer cell line (Chisamore, et al. Clin Cancer Res 2001; 7(10):3156-65). Furthermore 17β-estradiol (E2) inhibits tumor growth in vivo as well as inhibits colony formation when cells are grown in 3D Matrigel (Zhang, et al. Mol Cancer Res 2009; 7(4):498-510). Here we show that the Src kinase inhibitor PP2 (5 μM) significantly inhibited T47D/PKCα colony formation in 3D Matrigel, but was unable to inhibit proliferation in vitro. The combination of PP2 and E2 treatment in 3D Matrigel did not further decrease colony number when compared to either alone. This suggests that these compounds may be acting on the same pathway and that E2 may be inhibiting Src kinase activity. To examine the effect of E2 on Src in T47D/PKCα xenograft tumors we probed the phosphorylation state of Src on Tyr416, known to be necessary for its full kinase activity. Src is basally phosphorylated on Tyr416 in T47D/PKCα cells. Interestingly, E2 treatment in vivo resulted in a decrease in phospho-Src. Erk 1/2, a downstream target of Src, is basally phosphorylated in T47D/PKCα cells. E2 treatment also resulted in a decrease in phospho-Erk1/2, in accordance with decreased Src phosphorylation. A reduction in phosphorylated Src and Erk1/2 was not seen in vitro indicating that the extracellular matrix is necessary for E2 to elicit an inhibitory effect on these proteins. In this study, we report that E2 treatment in vivo inhibited Src phosphorylation in tamoxifen resisitant T47D/PKCα tumors. Additionally pharmacological inhibition of Src inhibited T47D/PKCα colony formation in 3D Matrigel. Taken together, our results suggest that a Src inhibitor may be a potential therapeutic option for patients with PKCα-overexpressing breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2289. doi:10.1158/1538-7445.AM2011-2289
Abstract 2294: The nuclear association among ERα – PR – AIB1 is modulated by progestins and antiprogestins in experimental breast cancer
Authors: Giulianelli Sebastian J. , Gorostiaga María A. , Lanari Claudia
Published: June 2012

The nuclear receptor coactivator AIB1 (SRC-3) is intimately associated with hormone action in estrogen receptor (ER)-positive breast cancer, and its overexpression has been associated with early resistance to endocrine therapy. In two models of ER+, progesterone receptor (PR)+ breast cancer, human T47D cells growing in vitro and C4-HD and C4-HI mouse mammary carcinomas maintained by syngeneic transplantations in BALB/c mice, we have shown that progestins induce a nuclear physical association between ERα and PR isoform A (PRA). We hypothesize that this interaction is necessary to induce progestin-induced cell proliferation. While both the pure antiestrogen ICI 182780 (ICI) and the antiprogestin RU486 inhibit tumor growth, only ICI blocked the ERα-PRA nuclear interaction induced by medroxyprogesterone acetate (MPA). RU486 showed an agonist-like effect stimulating the association between both receptors. The aim of this study was to investigate the interaction of ERα with PR isoform B (PRB) in both models and to evaluate the role of AIB1 in progestin-induced proliferation and in RU486-induced inhibition of cell proliferation. MPA (10nM) treatment induced an early increase in the nuclear co-localization between pSer118 ERα and pSer162 PRB in epithelial cells from the mouse model (p<0.001), and between total ERα with pSer162 PRB in T47D cells (p<0.001), as shown by confocal microscopy. By co-immunoprecipitation assays we corroborated that both PR isoforms interact with ERα in the cell nuclei. We also found that MPA increased the nuclear association of pSer294 PR and ERα with AIB1 in the murine epithelial tumor cells (p<0.001). These results were corroborated by co-immunoprecipitation in nuclear extracts from untreated or MPA-treated T47D cells. RU486 (10nM) induced similar ERα-PR interactions as MPA, but no association between PR and AIB1 was observed in RU486-treated cells. Our data support the hypothesis that a nuclear complex of ERα, both PR isoforms, and AIB1 mediates the proliferative effects of MPA. In addition, the results suggest that the lack of association of ERα-PR complexes with AIB1 may explain the antagonist effect of RU486 in these cells, supporting the hypothesis that the inhibitory effects of RU486 are related to genomic events.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2294. doi:10.1158/1538-7445.AM2011-2294
Abstract 2311: A novel kinase inhibitor of FADD phosphorylation chemosensitizes through the inhibition of NF-κB
Authors: Schinske Katrina , Nyati Shyam , Williams Terence , Johnson Timothy , Ross Brian , Tomás Ricardo Pérez , Rehemtulla Alnawaz
Published: June 2012

Fas-Associated protein with Death Domain (FADD) was originally reported as a pro-apoptotic adaptor molecule that mediates receptor induced apoptosis. Recent studies have revealed a potential role of FADD in NF-κB activation, embryogenesis, and cell cycle regulation and proliferation. Over-expression of FADD and its phosphorylation have been associated with the transformed phenotype in many cancers and is therefore a potential target for therapeutic intervention. In an effort to identify novel chemical entities that inhibit FADD phosphorylation, we screened an NCI diversity compound collection using a cell based assay recently developed in our laboratory for the detection of FADD-kinase activity (Khan, 2010). The screen resulted in the identification of NSC 47147 as a potent inhibitor of FADD phosphorylation. This compound was shown to be an inhibitor of CKIα, a previously described FADD kinase, with an IC50 = 1.5 µM. We demonstrate that inhibition of FADD phosphorylation by NSC 47147 results in a decrease in NF-κB activity, thereby potentially sensitizing tumor cells to chemotherapy. In support of this, treatment of lung cancer cells with NSC 47147 and cisplatin in culture as well as in a mouse xenograft model resulted in the synergistic induction of apoptosis and enhanced tumor control compared to either agent alone. These results provide evidence for the inhibition of FADD phosphorylation as a viable target for anti-cancer therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2311. doi:10.1158/1538-7445.AM2011-2311
Abstract 2312: Using multidimensional real-time cell profiling approach to identify novel inhibitors of key targets and pathways involved in cancer maintenance and progression
Authors: Ke Ning , Wang Xiaobo , Xu Xiao , Abassi Yama
Published: June 2012

A high throughput cell-based phenotypic screening method was developed using real time cell analysis system which is based on impedance sensor technology. Using compounds with known mechanisms, we had previously demonstrated small molecule compounds produce characteristic time-dependent cell response profiles (TCRP) which could be indicative of mechanism of action. The cellular-basis for the anti-mitotic TCRP has been extensively validated using both anti-mitotic compounds and siRNAs targeting mitotic machineries (e.g. Eg5, PLK1 & Mad2). In this study, we used the TCRP approach to screen a 119,595 compound library for novel anti-cancer agents. The screening was performed at a final concentration of 17.8 µM using a human non small cell lung cancer line, H1993. The TCRP was monitored for 48 hours after compound treatment and the data generated was analyzed using in-house developed software to perform clustering analysis. For each screening set, a panel of compounds with known mechanisms of action were used as positive control and also served as reference for TCRP-based clustering to predict mechanisms of action. After multi-dose confirmation, 239 hits with IC50 values less than 8.9 µM were subjected to TCRP analysis. In comparison with reference TCRP patterns, we indentified 26 hits corresponding to tyrosine kinase inhibitors, 9 hits as DNA damaging agents, 125 hits as anti-mitotic agents, and 28 hits as cytostatic agents. Detailed characterization of 117 putative anti-mitotic compounds with traditional p-H3 based methods demonstrates 113 of them (96.6%) mediated mitotic arrest, thus demonstrating the robust predictive nature of the approach. We also have validated the putative tyrosine kinase inhibitors in in vitro kinase assays. TCRP-based phenotypic screening displays a novel drug screening approach with great potentials for discovering new pathway-modulating anticancer agents.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2312. doi:10.1158/1538-7445.AM2011-2312
Abstract 2313: DDR1 receptor kinase pathway as an emerging target in cancer therapeutics
Authors: Kim Hyung-Gu , Mandinova Anna , Hwang So Young , Schreiber Stuart L. , Lee Sam W.
Published: June 2012

We have identified a novel p53 pro-survival signaling pathway involving up-regulation/activation of a p53 target gene DDR1 and its down-stream signaling, which can influence the balance between cell death and survival in a p53 dependent response. Unlike other p53 target genes that function as either cell cycle inhibitors or apoptosis-promoters, DDR1 kinase promotes cell survival by counteracting p53-mediated cell death/apoptosis. Furthermore, accumulating evidence suggest that aberrant signaling through DDR1 kinase is closely associated with various steps of tumorigenesis and carcinogenesis, although little is known about the molecular mechanism(s) underlying the role of DDR1 in cancer. We have identified TK0019 as a DDR1 tyrosine kinase inhibitor via the target-based small molecule array screening. TK0019 specifically targets DDR1 kinase and its novel targets involved in EMT (Epithelial-Mesenchymal transition) pathway such as Slug, AKT and vimentin, which is known to play a major role in tumor progression and metastasis development. In addition, TK0019 suppressed the activation of AKT prosurvival protein and of essential EMT pathway proteins involved in tumor migration/invasion. DDR1 kinase is highly expressed in a variety of human cancer and occasionally mutated in lung cancer and leukemia. TK0019 binds directly to N-terminal part (extracellular region of the receptor) of DDR1 protein and specifically inhibits DDR1 kinase activation in response to its ligand. TK0019 demonstrated antiproliferative activity against a range of DDR1-overexpressing cancer cell lines without high cytotoxicity in normal cells. TK0019 also exhibited the potent anti-growth properties even in EGFR targeted (erlotinib and gefitinib) drug-resistant cancer cells. TK0019 potently inhibited spontaneous formed breast cancer tumors and their associated metastasis in mice with no apparent adverse effects on normal organs. More particularly, the invention features a novel use of TK0019 for effective treatment of the variety human cancers by inhibiting tumor migration/invasion via the inactivation of EMT pathway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2313. doi:10.1158/1538-7445.AM2011-2313
Abstract 2323: Molecular modeling studies of the STAT3 homodimer:DNA complex
Authors: Husby Jarmila , Haider Shozeb M. , Zinzella Giovanna , Thurston David E. , Neidle Stephen
Published: June 2012

Signal Transducers and Activators of Transcription (STAT) are a group of latent cytoplasmic transcription factors in cytokine signaling, that transduce signals from the cell membrane to the nucleus, where they activate gene transcription. STAT activation by phosphorylation is mediated by growth factors or cytokines, inducing STAT:STAT dimer formation between two monomers via reciprocal phosphoTyr formation. A member of the STAT family, STAT3 (Figure 1), has been found to be expressed at elevated levels in a large number of diverse human cancers, thus STAT3 represents a validated target for anticancer drug design. Understanding the dynamics of the STAT3 dimer interface, accounting for both protein-DNA and protein-protein interactions, with respect to the dynamics of the latent, unphosphorylated STAT3 monomer, is important for designing potential small-molecule inhibitors of the activated dimer.

The results of molecular dynamics (MD) simulations of the activated STAT3 homodimer:DNA complex, and latent unphosporylated STAT3 monomer in explicit water have been studied. Careful analysis of the data obtained from MD simulations over a 35 ns time-frame has revealed the dynamics of the macromolecular complex at the atomistic level; how the transcription factor interacts with the DNA, the nature of the conformational changes, and the way function may be affected. Further analysis of the dimer interface, with a main focus on the protein-DNA interaction, including the water molecules, has revealed the key residues contributing to the recognition events involved in STAT3 protein-DNA interactions. Moreover, locations of the majority of mutations in the DNA-binding domain are found at the protein-DNA interface. In particular, we have been able to map out these mutations in detail and relate them to specific protein-DNA contacts. Their structural stability and importance will be described, along with a discussion of how this model is being used to assist in the discovery of novel small-molecule inhibitors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2323. doi:10.1158/1538-7445.AM2011-2323
Abstract 2331: Role of MMP expression in the effect of Plk4 on cell migration and invasion
Authors: Brashavitskaya Volha , Rosario Carla O. , Dennis James W. , Swallow Carol J.
Published: June 2012

Plk4 is a haploinsufficient tumor suppressor in mice (Ko et al, Nature Genetics, 2005). Plk4+/− mouse embryonic fibroblasts (MEFs) spontaneously immortalize in culture and become tumorigenic by passage 15 (Rosario et al, PNAS, 2010). We investigated the secondary genetic alterations associated with tumorigenicity.

Early passage (P3) Plk4+/+, P3 Plk4+/− and late passage (P15) Plk4+/− MEFs were compared by genome-wide expression array (Illumina). When differentially expressed genes were organized by biological function, increased cell proliferation and death (p=2.13E10-1.31E02, p = 2.52E07-1.14E02 respectively), and decreased cell motility (p = 3.74E07-1.27E02) in the P15 MEFs were predicted. An independent array analysis comparing tumorigenic (T) to nontumorigenic (NT) Plk4+/− MEFs showed a pattern of altered gene expression predictive of decreased motility in the T MEFs. These results suggested the hypothesis that Plk4 modulates cell motility. To directly test this hypothesis in functional assays we measured cell spreading, scratch-wound healing and transwell migration through Matrigel; in all these assays P3 Plk4+/− were inferior to P3 Plk4+/+ MEFs. We hypothesized that altered MMP expression was one mechanism for the difference observed in migration, and tested this by comparing MMP-13,-10 and -3 expression as shown in the Table. Plk4 status was associated with altered expression of MMPs; in particular, MMP-13 expression was confirmed to be significantly reduced in P3 Plk4+/− compared to P3 Plk4+/+ MEFs by RT-qPCR.

Our results indicate that Plk4 regulates cell motility and invasion. One potential mechanism is through upregulation of MMP-3,-10, and -13 expression. To further elucidate the molecular mechanisms of Plk4 related carcinogenesis we will next directly determine the effect of Plk4 on MMP activity and correlate this with migration and invasion.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2331. doi:10.1158/1538-7445.AM2011-2331