bluuebunny/crossref_metadata_2025 huggingface dataset:
Treatment-adjusted predisposition to second malignant neoplasms after a solid cancer in childhood: A case-control study
Authors: Guérin S. , Hawkins M. , Shamsaldin A. , Guibout C. , Diallo I. , Oberlin O. , Brugières L. , de Vathaire F.
Published: March 2020

21031

Background: Previous therapy, genetic susceptibility and the type of the first cancer are known to be associated with the risk of a second malignant neoplasm (SMN) among patients treated for a childhood cancer. The aim of this study was to investigate the independent role of the type of first malignant neoplasm in the onset of any SMN taking into account treatment and family cancer syndromes. Methods: A case-control study nested in a European cohort of 4,581 patients, treated for a solid cancer during childhood was conducted. 146 cases with a SMN and 417 controls were matched according to sex, age at first cancer, chemotherapy, radiotherapy, the local radiation dose received at the site of the SMN for the case and at the same site for the matched controls and follow- up. Results: A significantly increased risk of developing any SMN was observed after Hodgkin's lymphoma, retinoblastoma, soft tissue sarcoma orand a malignant bone tumor as the first malignant neoplasm (FMN), independently of initial treatment and family cancer syndromes. A significantly increased risk of developing a sarcoma was observed among patients who had developed a retinoblastoma, OR=5.9 (95%CI : 1.0–34) a malignant bone tumor, OR=11.9 (95%CI : 1.5–95), or a soft tissue sarcoma, OR=5.1 (95%CI : 1.4–19) compared to patients with a Wilm's tumor as the FMN. Conclusions: Survivors of Hodgkin's lymphoma, retinoblastoma, soft tissue sarcoma or a malignant bone tumor are at increased risk of any SMN independently of initial treatment and genetic susceptibility.

No significant financial relationships to disclose.
Single nucleotide polymorphisms in androgen-related genes and prostate cancer
Authors: Gos M. , Sadowska M. , Wiechno P. , Demkow T. , Janik P.
Published: March 2020

21033

Background: Single nucleotide polymorphisms (SNPs) in genes encoding proteins involved in androgen metabolism were suggested to influence the individual prostate cancer susceptibility and clinical course of the disease. To examine this correlation in Polish population, we have developed a case-control study to analyze whether SNPs in CYP17 (+34T/C), SRD5A2 (V89L and A49T) and UGT2B15 (D85Y) genes may influence prostate cancer risk. These SNPs were also examined for their correlation with clinical features of prostate cancer at diagnosis and disease progression to hormone-refractory state. Methods: The genomic DNA was extracted from blood samples from 182 men with histologically confirmed prostate cancer and 217 healthy men, randomly chosen from population. SNPs analyses were performed with standard molecular methods. Results: The case-control study has revealed that 85YY and 85DY variants in UGT2B15 were more prevalent among patients (83%) than in the control group (73.3%; p=0.02; OR=1.78). The +34CC genotype in CYP17 was more frequent in patients with distant metastasis (30.4%) than in patients with organ confined (T1/T2) or locally advanced (T3/T4) disease (10.2% and 22.6%, respectively; p<0.05). This variant also correlated with higher PSA level at diagnosis. The 89LL variant in SRD5A2 was more common in patients with poorly differentiated prostate cancer (Gleason >6; 17.8%) than in men with well differentiated tumor (5.4%; p=0.033, OR=3.78). The 85YY variant in UGT2B15 was more prevalent in patients that developed hormone-refractory prostate cancer within two years from the beginning of androgen blockade (47.2% vs 23.3%, p=0.025, OR=2.95). Also the analysis of progression-free survival time on hormonal therapy for D85Y polymorphism yield significant results (p=0.041, Gehan's generalized Wilcoxon test). Conclusions: The D85Y polymorphism in UGT2B15 seems to influence prostate cancer risk in Polish population. The analyzed SNPs also correlated with clinical stage and PSA level at diagnosis (+34T/C in CYP17), tumor histological grade (V89L in SRD5A2) and response to hormonal therapy (D85Y in UGT2B15).

No significant financial relationships to disclose.
Retrospective analysis of phosphorylation status of the estrogen receptor in patients with early stage disease
Authors: Swaby R. F. , Huang M. , Ruth K. J. , Ross E. A. , Gong Y. , Page R. E. , Freedman G. M. , Goldstein L. J. , Di Cristofano A.
Published: March 2020

21034

Background: The AKT kinase, when phosphorylated to its activated form (pAKT), mediates many proliferative and survival effects of growth factor receptors such as members of the epidermal growth factor receptor (EGFR) family. Overexpression of AKT in breast neoplasms suggests that AKT activation is involved in breast cancer development and/or progression, and contributes to poor outcome. pAKT is frequently upregulated in Her-2/neu positive breast cancers. Activated AKT is able to phosphorylate in vitro serine 167 (pS167) of estrogen receptor alpha (ERa), resulting in ER-mediated transcriptional activity in the absence of estrogen and possibly contributing to tamoxifen resistance in ERa positive breast cancers. In addition, ERa is phosphorylated at S118 (pS118) by MAPK. Thus, it is possible that pAKT- mediated ERa phosphorylation is implicated in tumorigenesis and resistance to treatment with hormonal therapy in patients with breast cancer. Methods: Utilizing commercially available antibodies, IHC analysis of 125 paraffin-embedded, ERa positive, early breast cancer cases, stratified for stage, were tested for pAKT, PTEN, pS167, and pS118. Staining intensity and number of positive cells were assessed by 4 independent graders. Fisher's exact test was used to determine the significance of associations with a two-sided type I error of 5%. Results: Phosphorylation of AKT occurred in approximately one-half of cases. pS167 was phosphorylated in 66% of pAKT positive cases compared to 35% of pAKT negative cases (p<0.0001). When we analyzed nuclear pAKT staining, this association was more pronounced: 76% of pAKT(+) vs. 37% of pAKT(-) (p<0.0001). 69% of pS118(+) cases were pS167(+) compared to only 15% pS167(+) when pS118 was negative (p<0.0001). In approx. 32% of cases pAKT was present in both the cytoplasm and nucleus. 20% of samples had only cytoplasmic pAKT, while in 4% of cases pAKT was nuclear only, supporting the hypothesis that cytoplasmic phosphorylation of AKT may precede nuclear accumulation. Conclusions: Phosphorylation of pS167ERa is significantly associated with AKT activation (especially in the nucleus) in early breast cancer, supporting the hypothesis that these two events may be causally linked and contribute to breast cancer pathogenesis.

No significant financial relationships to disclose.
Diffuse large B-cell lymphoma (DLBCL) in African American patients: A retrospective study of clinical, molecular, and therapeutic differences in a sample population treated at UAB Comprehensive Cancer Center (UAB CCC)
Authors: The A. S. , Reddy V. , Li Y. , Davis R. , Baird M. , Foran J. M.
Published: March 2020

21035

Background: SEER registry data indicate African Americans (AA) have a lower incidence of non-Hodgkin's lymphoma (NHL) but face higher lymphoma-specific mortality rates than Caucasians (C). To investigate this we compared outcomes between AA & C patients (pts) with DLBCL treated at UAB CCC. Correlative histologic & molecular studies of established prognostic features were performed to investigate differences. Methods: After IRB approval, DLBCL pts diagnosed '95-'06 were identified from pathology & referral databases. Baseline demographic & clinical data were extracted, including treatment (Rx), response & survival. Expression patterns of CD10, bcl-6, & MUM-1 were used to identify germinal center B-cell-like (GCB) vs non-GCB molecular subtype of DLBCL based on algorithm of Hans et al (2004). HLA-DR, bcl-2, & CD138 expression was determined. A 2:1 age- & gender-matched comparison of C to A pts was performed to evaluate differences. Descriptive analysis with X2 & Wilcoxon nonparametric tests & Kaplan Meier survival analysis were performed to determine differences. Results: 188 DLBCL patients were identified. Race was noted in 129, including 18 AA pts (14%). In comparison to 36 C pts (2:1 match), no differences were seen in LDH or stage at presentation. AA pts achieved better response to 1st-line Rx (p=0.01) & received fewer regimens (1 vs >1, p=0.05), however they were more likely to receive rituximab with 1st-line Rx (p=0.02), likely reflecting presentation in the post-rituximab era. Median survival, not yet reached for AA's, was 23 months for C pts (p=0.0509). Univariate & parametric survival analyses demonstrated LDH (p=0.0217) & 1st-line rituximab (p=0.0048) independently affected survival. In a separate cohort, no significant difference between races was seen in frequency of GCB vs non-GCB subtype or in expression of bcl-2, CD-138, or HLA-DR. Conclusions: Contrary to SEER observations, the outcome of AA DLBCL pts was superior to C pts in this single center study. No molecular or clinical prognostic feature dominated in either race. The fact that more AA than C patients received rituximab upfront confirms the benefit of adding this agent to 1st-line Rx.

No significant financial relationships to disclose.
Sarcomas predominate in mice deficient for PrPc and p53
Authors: Michael D. , Okuducu A. F. , Holtkamp N. , Bamme T. , Jehn C. F. , Riemer C. , Baier M.
Published: March 2020

21039

Background: For cellular prion protein (PrPc) putative roles in many cellular processes including signalling, survival, adhesion, proliferation and differentation have been proposed. The hallmarks of cancer include some of these traits and in human tumors both increased and decreased levels of PrPc have been observed. However PrPc deficient mice show no increased rate of tumor formation. To characterize the possible role of PrPc in tumorigenesis we generated PrPc-/-/p53-/- mice. Methods: Organ and tumor samples were surgically removed post mortem and processed for histological analysis. Time to death was defined as the latency between birth and spontaneous death or a terminal disease state as indicated by tumor burden and other symptoms of severe disease. Results: As reported previously lymphomas are the most frequent tumors in p53-/- mice [5 out of 7 (71 %)]. In contrast, sarcomas predominate in PrPc-/-/p53-/- mice [9 out of 21 (43 %)]. This was accompanied by a decrease in the incidence of lymphomas in PrPc-/-/p53-/- mice [7 out of 21 (33 %)]. Conclusions: These results indicate for PrPc, in the context of p53 deficiency, a role in sarcoma tumorigenesis and the PrPc-/-/p53-/- mice might provide a clinical relevant model for the study of sarcomas.

No significant financial relationships to disclose.
Prognostic value of immunohistochemical phenotypes (IP) of 141 operable breast cancer patients (pts) included in phase III trials of adjuvant therapy
Authors: Trujillo R. , Gallego E. , Márquez A. , Ribelles N. , Trigo J. , Perez D. , Quero C. , Medina A. , Vicioso L. , Alba E.
Published: March 2020

21040

Background: Gene expression arrays and IP studies classified breast cancer in three distinct subtypes: basal, HER2/neu and luminal that are associated with different clinical outcomes. Methods: In 141 pts with operable breast cancer, included in phase III trials of adjuvant therapy in our center, immunohistochemical staining was performed on 3μm sections of paraffin blocks, containing tissue-arrays of tumour tissue.A basal phenotype (BP) was defined by negative estrogen receptor (ER) and progesterone receptor (PR) and positive cytokeratin (CK) 5/6 or EGFR immunoreactivity. HER2/neu phenotype as positive c-erb B2 by HercepTest™ and luminal phenotype (LP) by positive ER, PR and CK 7/8 and negative HER-2. Survival curves were calculated by the Kaplan-Meier method. The differences between survivals were estimated using the log rank test. Multivariate Cox regression analysis was used to evaluate any independent prognostic effect of the variables on disease-free survival (DFS). Results: Complete clinical follow-up information was available for 141 pts. The median follow-up period was 52 months (range 1–103 months). During this period, 13.8% pts died from breast cancer and 27.7% pts relapsed. At the time of the primary diagnosis 10.4% of the pts had lymph node negative disease and 89.6% had positive lymph nodes. 50.8% pts received taxane chemotherapy, 7.7% Trastuzumab, 62.3% radiotherapy and 61% pts received hormonotherapy. Positivity for LP was 65.2%, BP 9.9% and Her-2 phenotype 8.5%. 16.3% didn't fit for any of the three subtypes. Median DFS for BP: 24 moths, for LP and Her-2 phenotypes median DFS was not reached. 5 years DFS were; BP: 19%, LP: 63% and Her-2: 56%. Kaplan-Meier survival analyses demonstrated that the presence of a detectable BP was highly significantly associated with a worse DFS compared with the presence of a LP, log rank test (p= 0.0001). Multivariate Cox regression analyses estimated that the prognostic effect of BP in relation to DFS was independent of lymph node, stage and tumor size, HR: 0.12 95% CI (0.05–0.2). Conclusions: We found that expression of BP was associated with poor prognostic in the context of randomized phase III trials.

No significant financial relationships to disclose.
Translation of gene expression profiles into a nine antibody IHC multivariate index assay for colorectal carcinoma
Authors: Schreeder M. T. , Seitz R. S. , Beck R. , Ring B. Z. , Ross D. T.
Published: March 2020

21042

Introduction: Pathologic stage is the most powerful predictor of outcome in colorectal cancer. However, in early stage disease there continues to be a need for better identification of patients at high risk of disease progression after surgical resection. We have undertaken a large scale effort translating gene expression profiles into immunohistochemical biomarkers that classify carcinoma in novel ways. Herein we report the identification of a colorectal tailored antibody “panel of diversity” that distinguishes tumor heterogeneity and propose a multivariate index assay (MIA) for predicting outcome. Methods: 744 stage I-III consecutive colon cancer patients treated by the CCIH between 1992 and 2000 were identified. Clinical followup through 2005 was obtained from the tumor registry and verified by chart review . Tissue micorarrays (TMA) were constructed from these patients resected primary tumors. Over 700 antibodies targeted by carcinoma gene expression experiments or literature review were first screened on a 30 case TMA. Of these, 34 were selected as showing subjectively high quality stains and the ability to classify patient populations. These antibodies were used to stain the clinical TMA and scored using a semi-quantitative scale. We used a number of approaches including Cox, RPART, and Bayesian to derive candidate MIA's to predict recurrence. Results: 13 antibodies showed a significant or near significant association with either overall recurrence or recurrence at 5 years. We have previously shown these antibodies to have similar prognostic abilities in other solid tumor carcinomas. We propose a model combining nine markers (p53, CCNA2, TFF3, RERG, AKR1C1, TLE3, IRX3, SYP, TTC7) as an MIA for predicting outcome in Stage II patients. The model is independent of pathologic stage. Conclusions: This colon tailored antibody ‘panel of diversity’ is a tool for characterizing the biologic diversity in colon cancer cohorts. The identification of nine prognostic markers that can be effectively combined using an MIA suggests that combination of multiple markers will be useful for developing sensitive and specific prognostic assays. The model proposed herein should be validated using additional cohorts to evaluate its overall clinical utility.

No significant financial relationships to disclose.
Prognostic value of HER-2/neu overexpression for the development of brain metastasis after treatment of early breast cancer
Authors: Ahn J. , Kim S. , Kim W. , Gong G. , Kim M. , Son B. , Ahn S. , Lee S. , Kim H.
Published: March 2020

21044

Background: Several studies have addressed the incidence of brain metastasis in HER-2/neu-overexpressing metastatic breast cancer. However, the relevance of HER-2/neu overexpression as a risk factor for brain metastases in breast cancer has not yet been clearly defined. The aim of this study is to analyze the risk of brain metastasis in HER-2/neu-positive early breast cancer patients in comparison with HER-2/neu-negative patients. Methods: We used tissue microarray to examine HER- 2/neu overexpression by immunohistochemistry (IHC) in 773 consecutive patients diagnosed with breast carcinoma who underwent surgery between January 1993 and December 1998. Patients were considered HER-2/neu-positive if they were 3+ on IHC testing. A total of 194 (25.1%) patients with HER-2/neu-overexpressing newly diagnosed breast cancer were identified in our study. The medical records and pathologic data of all 773 patients were reviewed retrospectively. Results: During a median follow-up period of 8.1 years, brain metastasis were identified in 7.2% (14 patients) with 194 HER-2/neu-overexpressing breast cancer compared with 2.6% (15 patients) in 597 HER-2/neu-negative patients (hazard ratio 2.924 [1.385–6.175], P=0.005). More HER-2/neu-overexpressing patients developed distant metastasis (33.5% vs. 18.8%, P=0.0001). The 8-year overall survival was 76.1% for HER-2/neu-negative patients versus 56.4% for HER-2/neu-positive patients (P<0.00001). By multivariate analysis, HER-2/neu overexpression, tumor size larger than 2 cm, at least one positive node, and progesterone receptor (PR)-negativity were independent risk factors for the development of brain metastases. Conclusions: Our data show that HER-2/neu overexpressing early breast cancer patients are at significantly increased risk for brain metastases. Also, our results demonstrate that HER-2/neu overexpression is an important independent prognostic factor of early breast cancer, and suggest that more aggressive adjuvant chemotherapy and radiologic screening to detect brain metastases should be considered in this population.

No significant financial relationships to disclose.
Stage and histology influence the relationship between MDM2 promoter polymorphism and esophageal cancer and overall survival (OS)
Authors: Mackay H. J. , Bradbury P. , Asomaning K. , Zhou W. , Kulke M. , Su L. , Suk Heist R. , Lynch T. J. , Christiani D. C. , Liu G.
Published: March 2020

21047

Background: A single nucleotide polymorphism in the MDM2 promoter (SNP309) has been found to affect OS of advanced stage gastric adenocarcinoma (AD) and early stage squamous (SQ) cell carcinoma of the lung. The aim of this study was to evaluate the role of this polymorphism in the prognosis of esophageal cancer, another aerodigestive cancer. Methods: 150 early stage (E) and 118 locally advanced stage (LA) esophageal cancers were genotyped for MDM2 SNP309 using Taqman. The primary endpoint was overall survival (OS). Results: E disease: n=23 stage I; n=127 stage II. LA disease: n=93, Stage III; n=25, Stage IVA. AD comprised 215 (81%), while SQ comprised 45 (17%) of cases; 8 (3%) had poorly differentiated tumors. Median follow-up = 32 months. Median OS were 36 and 21 months for E and LA disease, respectively. Both histology and disease stage affected the relationship between SNP309 and esophageal cancer OS (see Table ). The wildtype T/T genotype conferred a worse OS in E patients (log-rank, p=0.03), especially those with AD (log-rank, p=0.003). In Cox proportional hazards interaction analyses, after adjusting for age, gender, stage and PS, there were statistically significant interactions between MDM2 SNP309 and disease stage (interaction p=0.004) and between MDM2 SNP309 and histologic subtype (AD vs. SQ)(interaction p=0.02). Thus, the direction of SNP309 association from our AD and E esophageal cancer patients are opposite to those of our SQ and LA esophageal cancer patients. However, our SQ and LA results are similar to the SQ lung cancer and advanced stage gastric cancers previously reported. This suggests that biologic mechanisms underpinning the prognostic role of SNP309 are dependent on extent of disease and histologic subtype. Conclusion: Histology and disease stage interact with the prognostic role of MDM2 SNP309 polymorphism in esophageal cancer OS.

[Table: see text]

No significant financial relationships to disclose.
One Step Nucleic Acid Amplification (OSNA) for intra-operative detection of lymph node metastases in breast cancer patients
Authors: Schem C. , Maass N. , Bauerschlag D. O. , Hilpert F. , Roder C. , Löning T. , Carstensen M. , Jonat W. , Tiemann K.
Published: March 2020

21053

Background: Sentinel lymph node metastasis in breast cancer patients has conventionally been determined by intra- operative histopathological examination of frozen sections followed by definitive post-operative examination of permanent sections. The purpose of this study is to evaluate a more efficient method for intra-operative detection of lymph node metastasis in a clinical setting. Therefore, a new rapid method of detection of CK19 mRNA by One Step Nucleic Acid Amplification (OSNA) and Reverse Transcription Loop Mediated Isothermal Amplification (RT-LAMP) was tested against standard histological techniques. Methods: The clinical study was conducted at two facilities using 188 lymph nodes (46 patients). Lymph nodes from axillary dissected patients with breast cancer were divided into four slices. Alternate slices were used for the new method of CK19 mRNA detection (OSNA) and histopathological examination by hematoxylin & eosin and cytokeratin staining. For the analysis of concordance between both methods, the slices for histopathology were cut in 4 μm sections. For the OSNA method the other 2 slices were homogenised in a short sample preparation step. Afterwards CK19 mRNA was directly amplified from the tissue lysate within 16 minutes. Results: In the clinical study, an overall concordance rate between the OSNA assay and histopathology was 92%. Several discordant results are explainable with the expected uneven distribution of metastases within the lymph nodes. Conclusions: The study indicates that OSNA is a reliable and fast diagnostic tool for the detection of lymph node metastases in breast cancer patients. This method could be beneficial to avoid diagnostic delay for the patient or second surgery based on a post-operatively diagnosed positive lymph node.

No significant financial relationships to disclose.
The MUC16 mucin gene upregulates gene transcripts associated with invasion
Authors: Diaz J. P. , Kehoe S. , Thapi D. , Rosales N. , Ma X. , Yan X. J. , Spriggs D. R.
Published: March 2020

21057

Background: The MUC16 gene encodes the CA125 ovarian cancer antigen and may play a role in invasion. The aim of this study is to investigate the properties of MUC16 in vitro. Methods: PCR was used to create a DNA construct, MUC16–1R-GFP, containing all of the MUC 16 sequence proximal to the second cysteine loop repeat and a GFP-vector construct. SKOv-3 and T-80 cell lines were selected because they are negative for immunologically active CA125. Transfections were performed utilizing the phrGFP II C vector system. Stable transfectants were sorted for GFP by FACS and were grown as populations. The growth rate of the transfected populations were compared to vector only. Periodically the stable transfectants were analyzed by FACS for GFP and OC125 mAb. The in vitro invasive potential was examined, using a Matrigel® invasion assay. RT PCR was used to examine the effect of MUC16 constructs for a panel of invasion focused genes. Immunoblotting was performed on the 1R protein and probed for all genes that were significantly changed on RT PCR. Results: The SKOv-3 1R transfected cell lines produce measurable CA125 into the cell culture medium (4–10 U/mL) while T-80 1R cell lines did not. Analysis of the in vitro growth rate did not demonstrate any difference between the transfectant and vector only cell lines. The 1R-GFP transfected lines showed MUC16 cell surface expression by FACS analysis. Invasion assay of the MUC16 transfectants demonstrated substantially more invasive potential than vector only (SKOV-3: 36% vs. 15% invasion; T-80: 35% vs. 23% invasion). Several transcripts associated with invasion and metastasis were up-regulated 3–9 fold including CDH1, FN1, IL1B, MMP7 and MMP9. Transcripts for the tyrosine kinases SYK and HTATIP2 were down-regulated 16–20 fold respectively. Western immunoblotting confirmed the RT PCR findings. Conclusions: The1R-GFP construct did not alter the growth rate of transfectants in vitro; however; the SKOV-3 1R and T-80 1R cell line did have substantially more invasive potential than vector only. Five transcripts associated with invasion were up-regulated, and 2 transcripts for kinases were down-regulated in the transfected cell lines. These data supports an association between MUC 16 and invasive potential.

No significant financial relationships to disclose.
Transforming genomic grade index (GGI) into a user-friendly qRT-PCR tool which will assist clinicians and patients in optimizing treatment of early breast cancer (BC)
Authors: Durbecq V. , Toussaint J. , Haibe-Kains B. , Desmedt C. , Rouas G. , Larsimont D. , Bontempi G. , Buyse M. , Piccart M. , Sotiriou C.
Published: March 2020

21058

Background: We have recently shown that proliferation captured by the genomic grade index (GGI) is the most important prognostic factor in BC, far beyond estrogen receptor (ER) status and may encompass a significant portion of the predictive power of many previously published prognostic signatures. Furthermore, we have demonstrated that GGI can identify two clinically distinct ER-positive molecular subtypes in a simple and highly reproducible manner across multiple datasets. The aims of this study were to convert and validate by qRT-PCR assay the prognostic value of the GGI using frozen (FS) and paraffin-embedded tumor samples (FFPE) from early BC patients. Methods: We first developed a qRT-PCR assay based on 8 selected GGI genes involved in different phases of the cell cycle and 4 reference genes. We then tested its accuracy and concordance with the original microarray derived GGI using a BC population from which FS, FFPE tissues and microarray data were available (N=30). Finally, we assessed its prognostic value on an independent ER-positive tamoxifen only treated BC population (N=82). Results: A statistically significant correlation was observed between GGI generated by microarray and qRT-PCR assay using FS material (r=0.91; p<0.0001) as well as between GGI using qRT-PCR derived from FS and FFPE tumor samples (r=0.86; p<0.0001). A high GGI levels assessed by qRT-PCR (GGRI) was associated with a higher risk of recurrence in the ER-positive tamoxifen only treated patients [HR= 2.319 (95% CI: 1.245–4.321), p=0.008] in accordance with our previous microarray results. Interestingly, in a multivariate analysis, GGRI remained significant [HR=1.956 (95% CI: 1.031–3.710), p=0.04] together with age (<50y) and tumor size (>2cm). Conclusions: In this study, we report a qRT-PCR assay based on a limited number of genes, which recapitulates in an accurate and reproducible manner the prognostic power of the microarray derived GGI using both FS and FFPE tumor samples. Further validation of its prognostic value is currently ongoing on two independent populations totaling over 500 BC patients.

No significant financial relationships to disclose.
Lack of p21/Waf-1 expression and lymphocyte infiltration in primary ovarian cancer correlate with the somatic HLA-A2 expression
Authors: Andersson E. , Villabona L. , Kanter L. , Gamzatova Z. , Kiessling R. , Dalianis T. , Bergfeldt K. , Masucci G. V.
Published: March 2020

21059

Background: HLA-A2 allele is highly prevalent in Scandinavia and decreases with latitude in Europe. Similarly, ovarian cancer mortality decreases with latitude in European countries. We have previously shown that HLA-A2 is associated to a severe prognosis in serous adenocarcinoma of the ovary in stage III-IV. Others have shown that lack of p21/Waf-1 and lymphocyte infiltration in this tumour is correlated to poorer prognosis as well. Here we analyse a possible linkage between these variables. Methods: Thirty eight Swedish women with epithelial ovarian cancer were analyzed for the HLA-A genotype locus by PCR/sequence-specific oligonucleotide hybridization procedure (PCR/SSOP). Tissue slides from paraffin embedded tumour material were stained with haematoxylin and antibodies anti-p21 and -p53. Results: Twenty two patients were HLA-A2 positive. Fourteen of them (37%) lacked lymphocyte infiltration wich differed significantly from HLA-A2 negative patients (n=4, 11 %) (p=0.002). All of the HLA-A2 positive patients, with serous adenocarcinoma that showed absence of tumour lymphocyte infiltration, did not express p21/waf-1 (n=7 18 %). No cases in the HLA-A2 negative group with the same parameters were identified (p=0.009). Conclusions: These observations identify a high risk group of patients by linking together the somatic expression of HLA-A2 to the lack of lymphocyte infiltration and absence of p21/Waf-1 expression at the tumour level as prognostic negative factors. The information might have a relevant impact on the patient management.. In order to obtain a stronger power the analysis will be extended to a larger group of patients.

No significant financial relationships to disclose.
Interval and non-compliant breast cancers diagnosed in women aged 50–69 in a Canadian breast cancer screening program: Clinical-pathologic correlations
Authors: Rayson D. , Abdolell M. , Payne J. , Foley T. J. , Younis T. , Barnes P. J. , MacIntosh R. F. , Caines J.
Published: March 2020

21060

Background: Assessment of interval breast cancers and screening compliance are important parameters in the assessment of any comprehensive breast screening program. Clinical-pathologic examination of interval cancers may also be helpful in understanding differences between screen-detected and interval disease. The Nova Scotia Breast Screening Program (NSBSP) was founded in 1991 and, as of Jan 2007, 86,071 women aged 50–69 have entered the program, with 255,350 screening examinations completed. Methods: The Mammography Information System and Diagnostic Reporting System databases were used to identify all interval (diagnosed after a negative screen and before the recommended next screen) and non-compliant (diagnosed after a negative screen and after the recommended next screen) invasive breast cancer diagnoses. Unpaired t-tests were used to compare age at diagnosis, detection time and tumor size and the Chi- square test was used to compare the odds ratio of nodal involvement between interval and non-compliant cases. Results: A total of 1,189 screen-detected, 309 interval and 260 non-compliant invasive breast cancers were diagnosed amongst program participants aged 50–69 over this time period. Mean values (standard deviation, SD) for interval vs. non-compliant cases were; age at diagnosis 58.8 (SD=5.6) vs. 59.6 (SD=5.9) years (p=0.113); time to detection from last screen 363.9 (SD=187.6) vs. 1296 (SD=868.4) days (p<0.0001); tumor size 15.9 (SD=15.0) vs. 14.6 (SD=14.7) mms. (p=0.342). The incidence of nodal involvement in interval vs. non-compliant breast cancers was 32% and 23.5% respectively resulting in an odds ratio of 1.54, p=0.023. Conclusions: Interval and non-compliant breast cancers made up a significant proportion of all invasive cancers diagnosed amongst program participants. The finding of more cases having involved nodes at diagnosis despite no significant difference in mean age at diagnosis or tumor size suggests that interval disease may have a more aggressive phenotype. Data on tumor grade, lymphovascular invasion, hormone receptor status and HER2/neu over-expression are being collected.

No significant financial relationships to disclose.
Expression of a cancer associated isoform of PCNA in breast cancer has implications as a potential biomarker
Authors: Malkas L. H. , Schnaper L. A. , Herbert B. , Abdel-Aziz W. , Liu Y. , Dobrolecki L. E. , Hoelz D. , Agarwal B. , Badve S. , Goulet R. J. , Hickey R. J.
Published: March 2020

21069

Background: We have identified a novel cancer associated isoform of the protein, proliferating cell nuclear antigen (PCNA), termed (caPCNA), and demonstrated that this isoform arises through a direct protein post-translational modification, and not via genetic mutation or RNA splice variation. Our results suggest that caPCNA has the potential to serve as a highly effective and unique marker for identifying malignant breast cancer cells. Methods: This assertion is based on our proteomic-based analyses of more than 60 malignant and non- malignant breast cell lines and tissues. Commercially available antibodies against PCNA cannot distinguish between the different isoforms of PCNA present in malignant and non-malignant breast cells and cannot be used clinically to differentiate between normal and malignant breast tissue. However, we have recently developed a rabbit polyclonal antibody, (caPCNAab), which specifically recognizes only the caPCNA isoform expressed by malignant human breast cells. Results: Using this antibody we clearly show that caPCNA is expressed only in malignant breast cells and tissues, and can be found in early disease. caPCNA expression in tissues was quantified as average staining intensity X average percentage of cells stained. Using this criterion the following data were obtained: 10 cases of normal breast tissue (reduction mammoplasty) gave a total score of 1%; 35 cases of normal breast tissue adjacent to malignancy scored as 4%; 30 cases of DCIS scored as 90%; and 55 cases of invasive breast carcinoma scored as 120%. The five cases of ADH examined thus far were shown to score similar to that of normal breast tissue. Conclusions: The implication of these data is that the development of a caPCNAab-based IHC stain could potentially be used to reliably stain only in situ or invasive carcinoma, and distinguish genuinely benign lesions (e.g., ADH) from carcinoma, (e.g., DCIS) allowing definitive diagnosis in such cases where a limited amount of an atypical lesion prevents definitive diagnosis on routine H/E stained sections alone. For the patient population, this could result in a marked decrease in the need for either a repeat core biopsy or an excisional biopsy due to an inconclusive initial diagnosis.

No significant financial relationships to disclose.
Epidermal Growth Factor Receptor (EGFr) status detection in correlation to objective response on cetuximab-based therapy in patients (pts) with advanced colorectal cancer (ACC)
Authors: Gevorgyan A. , Di Bartolomeo M. , Andreola S. , Orsenigo M. , Casieri P. , Signoroni S. , Cortinovis D. , Martinetti A. , Pilotti S. , Bajetta E.
Published: March 2020

21070

Background: In current clinical practice Cetuximab represents a standard therapy for Irinotecan resistant pts with ACC, where EGFR-immunohistochemistry (IHC) evaluation is positive. In absence of reliable molecular predictors of the clinical benefit to anti-EGFR therapy, we have studied EGFR protein expression and gene status in order to correlate them to the clinical response. Methods: We investigated 40 primary colorectal carcinoma specimens. EGFR-protein expression was evaluated by Immunohistochemistry (IHC) using PharmDX-Kit DAKO Cytomation. IHC evaluation was performed using combined score based on the sum of percentage of positive cells and the staining of each sample. We identified three EGFR-score categories: 0–2, 3–5, 6–7 corresponding to low, intermediate and high expression respectively. EGFR-genetic status was conducted by Dual-Colour FISH (LSI EGFR-probe Vysis Inc).Gene and chromosome 7 copy numbers were identified by fluorescence in situ hybridization (FISH-LSI EGFR-Dual-colour-probe Vysis-Inc) as follows: tumor samples had a high EGFR-gene copy number if there was high polysomy ( 4-copies 40% of cells) or gene amplification (gene-clusters, gene/chromosome ratio per cell of 2, 15 copies of EGFR/ cell in 10% of cells) . The analysis of clinical data from all patients treated with Cetuximab (400–250mg/m2/w) and Irinotecan (300mg/m2/d1q21) was then performed. Results: EGFR-(IHC) high score was observed in 7pts from which only 2 had an objective response (OR). Intermediate (13pts) with 3 OR, low (12pts) with 1 OR and negative (8pts) with 2OR. Main FISH patterns were: high polisomy/amplification in 8pts with no response on cetuximab therapy. Low polisomy described in 19pts with 3 cases of disease remission. Disomy in 13pts with 5OR. No relationship was detected between IHC-score evaluation and FISH pattern. The clinical benefit (OR+stable disease =6months) was 65% in score-group 0–2; whereas 46% and 57% in groups 3–5/6–7 respectively, without statistical significance. Conclusion: According to our experience, IHC and FISH are unable to provide adequate selection criteria for the patients with different EGFR-status expression, who can benefit from Cetuximab therapy.

No significant financial relationships to disclose.
SK3 polyglutamine tract polymorphism and acute oxaliplatin neurotoxicity: A translational study
Authors: Basso M. , Spada D. , Modoni A. , Cassano A. , Inno A. , Bagalà C. , Pozzo C. , Schinzari G. , Tonali P. A. , Barone C.
Published: March 2020

21076

Background: Acute neurotoxicity of l-oxaliplatin (l-OHP), characterized by cold-induced paresthesias, muscle cramps and tightness, seems electrophysiologically related to an abnormal nerve hyperexcitability (NHE) resembling neuromyotonia, an autoimmune disease caused by autoantibodies directed against voltage-gated potassium channels (VGKC). This l-OHP-related channelopathy seems to be reversible, but there is no agreement about the channel involved, most investigators favouring voltage-gated sodium channels. SK3 VGKC are mainly located in the peripheral nervous system and are characterized by an highly polymorphic CAG motif. We hypothesized that SK3 potassium channel dysfunction might be responsible for nerve hyperexcitability in patients treated with l-OHP. Methods: Pts eligible for an l-OHP-containing regimen were enrolled. Detailed neurological examination, nerve conduction studies (NCS) and needle electromyography (EMG) were performed before and after l-OHP administration. Furthermore, a venous blood sample was obtained from each pt in order to perform genetic analysis. Genomic DNA was extracted from pts’ leukocytes and PCR-mediated amplification of the second polyglutamine CAG tract of SK3 gene was carried out; PCR products were analyzed on polyacrylamide gel stained with Ethidium Bromide. Results: We evaluated 20 pts (14M, 6F); mean age was 59 yrs. According to neurophysiologic data it was possible to divide pts into 3 groups: G0 (no symptoms or signs of NHE), 7 pts; G1 (cold induced paresthesias-mild NHE), 8 pts; G2 (muscle cramps-severe NHE), 5 pts. Genetic analysis showed different alleles ranging from 125 to 225 bp. In particular, 75% of pts carrying a 175 bp allele experienced severe acute toxicicy (G2 group), and 15% mild toxicity (G1 group). The 175 bp allele was found only in 1/7 G0 pts (15%); in these pts the alleles distribution was diffusely scattered. Conclusions: Acute and reversible NHE observed in some pts treated with l-OHP might be produced either by an increased sodium conductance or by a decreased activity of VGKC. The high percentage of the 175 bp allele, that we found in the G2 group, induces us to think that the polymorphism at CAG locus of SK3 may be responsible for l-OHP toxicity. Sequence analysis of PCR gene products is ongoing.

No significant financial relationships to disclose.
The impact of DNA methylation on the identification of recurrent prostate cancer
Authors: Alumkal J. J. , Zhang Z. , Humphreys E. B. , Bennett C. , Mangold L. A. , Carducci M. A. , Partin A. W. , Garrett-Mayer E. , DeMarzo A. M. , Herman J. G.
Published: March 2020

21086

Purpose: Biochemical (PSA) recurrence of prostate cancer following radical prostatectomy remains a major problem. Better biomarkers are needed to identify high and low-risk patients. DNA methylation of promoter regions leads to gene silencing in many cancers. In this study, we assessed the impact of changes in DNA methylation on biochemical recurrence in men with prostate cancer. Methods: We examined the methylation status of fifteen genes using MSP (Methylation Specific PCR) on tissue samples from 151 patients with clinically localized prostate cancer for whom at least five years of follow-up after prostatectomy was available. Results: In a multivariable logistic regression analysis, extra capsular penetration, high Gleason score, and involvement of the lymph nodes, seminal vesicles, or surgical margin were associated with an increased risk of recurrence. In addition, samples with methylation of 2 specific genes involved in cell-cell adhesion and apoptosis were associated with biochemical recurrence with an odds ratio of 5.64 (95% CI=1.47–21.7, p=0.012) compared to samples without methylation of both of these genes. The methylation status of these 2 genes had a higher sensitivity (72.3%; 95% CI=57–84.4%) for detecting recurrences than all the clinico-pathological variables (p<0.02) except extra-capsular penetration (p=0.346). The methylation status of these 2 genes had a similar negative predictive value (79.0%; 95% CI=66.8–88.3%) as the individual clinico-pathological variables examined. Conclusion: DNA Methylation of specific genes is independently associated with an increased risk of biochemical recurrence after radical prostatectomy even one considers the prognostic clinico-pathologic variables used in the clinic today. Our findings should be validated on another larger group of patients with prostate cancer who have undergone radical prosatetectomies.

No significant financial relationships to disclose.
Molecular analysis of the Epstein-Barr virus LMP1 in Hodgkin lymphoma from Argentina and Brazil: Identification of a distinct LMP1 deleted variant
Authors: Chabay P. , Hassan R. , Guiretti D. , Valva P. , Barros M. , Lorenzetti M. , De Matteo E. , Zalcberg I. , Rey G. , Preciado M. V.
Published: March 2020

21099

Background: Epstein Barr virus (EBV), a human oncogenic virus, has two types, EBV1 and 2, and several polymorphic markers allow individual variants to be distinguished. Variations in EBV latent membrane protein 1 (LMP1) gene could define a more oncogenic strain or reflect geographic EBV origin. By sequence analysis of C-ter, N-ter and promoter (prom) regions, 4 distinct EBV groups (A to D) have been defined. Aim: To analyze LMP1 molecular variability of two different groups of Hodgkin`s lymphoma (HL) from the same geographic area. Methods: EBV association by EBERs in situ hibridization and LMP1 immunohistochemistry was analyzed in 27 HL from Argentina (Arg) and 36 HL from Brazil (Br). EBV type was assessed by EBNA PCR. C-ter LMP1 PCR was done in all cases. Sequencing of C-ter, N-ter and prom regions was done in 16 patients. Results: 21/27 (78%) Arg HL and 31/36 (86%) Br HL were EBV1, while 6/27 (22%) and 4/36 (11%) were EBV2, respectively (p= 0.29). Coinfection was observed in 1 (3%) Br HL. LMP1 deleted (del) variant was observed in 20/27 (74%) Arg HL and 20/36 (55%) Br HL (p= 0.18), as well as in non-neoplastic controls, 4/11 and 3/10 (36 and 30%) from Arg and Br respectively. Most LMP1 del displayed high number (5–7) of 33bp repeats (86% LMP1 del Br HL and 77% LMP1 del Arg HL) compared with LMP1 wt that exhibited low number (3–4) 33bp repeats (68% LMP1 wt Br HL and 100% LMP1 wt Arg HL). Analysis of LMP1 sequences showed that: LMP1 wt C-ter regions had unmutated N-ter and prom as B95.8, A and B groups (31%), except for a case showing new mutations. LMP1 del C-ter regions showed molecular identity with C group, but they showed new, undescribed mutations in prom and N-ter (63%). Conclusions: We found high frequency of LMP1 del variants in HL from Argentina and Brazil, which was associated with high number of 33bp repeats in C-ter region. This suggests a role for this variant in lymphomagenesis. A new molecular variant with characteristic promoter and N-ter mutations was identified in LMP1 del cases, which could be proposed as a regional South American variant.

No significant financial relationships to disclose.
EGFR is frequently hyperexpressed in human malignant peripheral nerve sheath tumors (MPNST) and is preferentially localized in high grade areas
Authors: Emile J. , Tabone S. , Bahleda R. , Terrier P. , Bieche Y. , Beauchet A. , Terrier-Lacombe M. , Lemoine A. , Penault-Llorca F. , Le Cesne A.
Published: March 2020

21105

Background: MPNST are highly malignant Schwann cell neoplasms that are generally resistant to conventional radiation and chemotherapy. Mouse models and in vitro data suggest that epidermal growth factor receptor (EGFR) expression is implicated in malignant transformation of Schwann cells. Methods: MPNST samples from 52 patients were studied for EGFR, Ki67, neurofibromin, p53 and survivin expression by immunohistochemistry and for EGFR amplification by in situ hybridization. Results were correlated with clinical data. EGFR RNA were also quantified by RT-PCR in 20 other MPNST and 14 cutaneous neurofibroma samples. Results: Half of the patients had a neurofibromatosis type 1 (NF1), and their age was lower at diagnosis (19 versus 27 years, P=0.04). EGFR expression was detected in 86% of MPNST, and was more frequent in NF1 (95% versus 75%). The staining of tumor cells was heterogeneous in several cases, and closely associated with high grade, Ki67 high and p53 positive areas. MPNST expressed a higher level of EGFR transcripts than neurofibromas. The five year survival was 33%, and NF1 status was the only prognostic factor in multivariate analysis. Conclusion: Our data suggest that EGFR may become a target for new therapies in MPNST, in particular in NF1-associated MPNSTs.

No significant financial relationships to disclose.
Detection of ovarian cancer-specific gene by differentially expressed gene polymerase chain reaction prescreening and direct DNA sequencing
Authors: Kim J. , Pak J. H. , Choi W. H. , Kim J. Y. , Joo W. D. , Kim D. Y. , Suh D. S. , Kim Y. M. , Kim Y. T. , Nam J. H.
Published: March 2020

21106

Background: To detect the genes differentially expressed in the ovarian cancer, we analysed the genes in the ovarian cancer and normal ovary by differentially expressed gene(DEG) PCR using the RNA extracted from the both tissues. We examined the relationship between the specific genes of ovarian cancer and pathogenesis of ovarian cancer. Methods: Differentially expressed genes were screened by ACP-based PCR. Differentially expressed bands were extracted from agarose gel, and then directly sequenced. Finally we determined the clinical importances of differentially expressed genes. Results: Some genes were overexpressed in the ovarian cancer tissue than normal ovary, such as plexin B1(PLXNB1), aminoacylase 1(ACY1), solute carrier family 25 protein(SLC25A5), triosephosphate isomerase 1(TPI 1), poliovirus receptor-related 3 protein(PVRL 3), clusterin, LY6/PLAUR domain containing 1 protein(LYPDC 1). And other five genes were more expressed in the normal ovary than ovarian cancer, such as ribosomal protein L11 and L23, tenascin XB (TNXB), complement component 1 and actin alpha 2. Conclusions: Clusterin was highly expressed in the tissue from ovarian cancer, which was identified with anti- or proapoptotic activity regulated by calcium homeostasis in prostate, breast and colorectal cancers. And it suggests the possibility that regulation of clusterin activity provides the prospect of breaking down cancer cells‘ resistance to apoptosis in the ovarian cancer. Ribosomal protein L11 and L23 was highly expressed in normal ovary, which plays an important role in regulating the stability and function of the p53 tumor suppressor protein. It suggests that suppression of ribosomal protein L11 may act an important role in proliferation of ovarian cancer and over-expression of ribosomal protein L11 may act an important role in cell cycle arrest in the treatment of the ovarian cancer.

No significant financial relationships to disclose.