Favorable Clinicopathological Features Research Articles

Doublecortin-like kinase 1 expression associates with breast cancer with neuroendocrine differentiation.

Doublecortin-like kinase 1 (DCLK1), a microtubule associated kinase, has recently been proposed to be a putative marker for stemness and adverse prognosis in gastrointestinal cancers. However, it is not clear whether the protein also plays similar roles in breast cancer. Here, the expression of DCLK1 was analyzed in a large cohort of invasive breast cancers (IBC) by immunohistochemistry. DCKL1 was associated with favorable clinico-pathologic features, namely lower histologic grade, absence of lymphovascular invasion, fibrotic focus, necrosis and lower pN stage (p≤0.045). Additionally, independent significant correlations were found with estrogen receptor and neuroendocrine markers (p ≤0.019), implicating its relationship with IBC with neuroendocrine differentiation (IBC-NED). In the current cohort, IBC-NED showed worse outcome than luminal cancers without NED (hazard ratio=1.756, p=0.041). Interestingly, within the IBC-NED group, DCLK1 was found to be a good prognostic factor (hazard ratio =0.288, p=0.011). These findings were in contrast to those in gastrointestinal cancers, suggesting different functional roles of DCLK1 in different types of cancers. In clinical practice, NED is not routinely assessed; thus IBC-NED are not well studied. Its poor outcome and significant heterogeneity warrants more attention. DCLK1 expression could aid in the prognostication and management of this special cancer subtype.

Characterization of HER2 Gene/Protein and Ki67 Protein Expressions in Colorectal Carcinoma Variants With Relation To Clinicopathological Parameters and Prognosis: An Immunohistochemical and Fluorescence In Situ Hybridization Study

Objective The data on the frequency and pattern of HER2 expression in colorectal carcinoma (CRC) and its clinical signifi¬cance are ambiguous. In addition, little is known about HER2 status in CRC variants and its relation with proliferative activity and clinical outcome. Such knowledge may be of potential value for therapeutic decision making in CRCs. Methods The HER2 gene/protein status was assessed by fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) in a tissue microarray of 150 CRCs and correlated with the expression of the proliferation marker Ki67, clinicopathological factors, and prognosis. Results CRCs were categorized into conventional adenocarcinoma (CA), 47 cases; adenocarcinoma with mucinous component (AMC), 28 cases; mucinous adenocarcinoma (MA), 56 cases; and signet ring cell carcinoma (SRCC), 19 cases. Compared to other variants, CA was significantly associated with favorable clinicopathological features, higher overall survival (OS) and disease-free survival (DFS), while SRCC and MA were significantly associated with ominous clinicopathological features, lower OS and DFS. Cytoplasmic HER2 overexpression was detected in 14.2% of CRC cases and showed a significant agreement with gene amplification. HER2 overexpression was significantly associated with favorable clinicopathological features notably early stages. High Ki67 expression was detected in 48% of CRC cases. HER2 and Ki67 were significantly different among CRC variants with AMC showing the greatest frequency of HER2 overexpression and Ki67 high expression than the other variants. The interrelation between HER2 and Ki67 expression was statistically insignificant and neither had any significant relation to OS or DFS in any of the CRC variants. Conclusions We conclude that mucinous histology infers an adverse prognosis in CRC. A subset of early stage CRC patients, with HER2 overexpression and possibly a distinct variant, may benefit from HER2 targeted therapy. IHC can be used as a method for screening of HER2 gene amplification in CRCs.

CD73 Predicts Favorable Prognosis in Patients with Nonmuscle-Invasive Urothelial Bladder Cancer.

Aims. CD73 is a membrane associated 5′-ectonucleotidase that has been proposed as prognostic biomarker in various solid tumors. The aim of this study is to evaluate CD73 expression in a cohort of patients with primary bladder cancer in regard to its association with clinicopathological features and disease course. Methods. Tissue samples from 174 patients with a primary urothelial carcinoma were immunohistochemically assessed on a tissue microarray. Associations between CD73 expression and retrospectively obtained clinicopathological data were evaluated by contingency analysis. Survival analysis was performed to investigate the predictive value of CD73 within the subgroup of pTa and pT1 tumors in regard to progression-free survival (PFS). Results. High CD73 expression was found in 46 (26.4%) patients and was significantly associated with lower stage, lower grade, less adjacent carcinoma in situ and with lower Ki-67 proliferation index. High CD73 immunoreactivity in the subgroup of pTa and pT1 tumors (n = 158) was significantly associated with longer PFS (HR: 0.228; p = 0.047) in univariable Cox regression analysis. Conclusion. High CD73 immunoreactivity was associated with favorable clinicopathological features. Furthermore, it predicts better outcome in the subgroup of pTa and pT1 tumors and may thus serve as additional tool for the selection of patients with favorable prognosis.

Androgen Receptor Expression Shows Distinctive Significance in ER Positive and Negative Breast Cancers

Androgen receptor (AR), a nuclear steroid hormone receptor, is differentially expressed in breast cancer subgroups with distinct clinical implications. To investigate the clinical significance of AR in breast cancers more precisely, the expression of AR in a large cohort of breast cancer was correlated with clinicopathological features, biomarker expression, and patients' survival according to different molecular groupings in this study. Higher AR expression was found in ER+ (57.8 %) than in ER- (24.7 %) cancers. In the ER+ cancers, AR expression was associated with favorable clinicopathological features, including lower grade (p < .001), lower pT stage (p < .001), and positivity for PR (p < .001). It was an independent prognostic factor for longer disease-free survival, mainly in the HER2+ luminal B cancers (hazard ratio [HR] = 0.251, 95 % CI 0.065-0.972, p = .045). In ER- cancers, AR expression was associated with features distinct from basal-like breast cancer, and such features were found in molecular apocrine (MA) cancers. AR correlated with presence of extensive in situ component (p = .006) and apocrine phenotype (p < .001), HER2 (p = .026), and EGFR (p = .048), but negatively with c-kit (p = .041), CK5/6 (p < .001), CK14 (p = .002), and αB-crystallin (p = .038). However, AR expression was found only in 37.8 % of immunohistochemically defined MA. Of note, AR-MA appeared to have a trend of worse overall survival than AR+MA. AR expression was different in ER+ and ER- cancers and had different clinical implications. AR alone may not be a good marker for MA subtype. Its expression in MA may have substantial prognostic implication and as such warrants further validation.

Analysis of copy number changes on chromosome 16q in male breast cancer by multiplex ligation-dependent probe amplification

Gene copy number changes have an important role in carcinogenesis and could serve as potential biomarkers for prognosis and targets for therapy. Copy number changes mapping to chromosome 16 have been reported to be the most frequent alteration observed in female breast cancer and a loss on 16q has been shown to be associated with low grade and better prognosis. In the present study, we aimed to characterize copy number changes on 16q in a group of 135 male breast cancers using a novel multiplex ligation-dependent probe amplification kit. One hundred and twelve out of 135 (83%) male breast cancer showed copy number changes of at least one gene on chromosome 16, with frequent loss of 16q (71/135; 53%), either partial (66/135; 49%) or whole arm loss (5/135; 4%). Losses on 16q were thereby less often seen in male breast cancer than previously described in female breast cancer. Loss on 16q was significantly correlated with favorable clinicopathological features such as negative lymph node status, small tumor size, and low grade. Copy number gain of almost all genes on the short arm was also significantly correlated with lymph node negative status. A combination of 16q loss and 16p gain correlated even stronger with negative lymph node status (n=112; P=0.012), which was also underlined by unsupervised clustering. In conclusion, copy number loss on 16q is less frequent in male breast cancer than in female breast cancer, providing further evidence that male breast cancer and female breast cancer are genetically different. Gain on 16p and loss of 16q identify a group of male breast cancer with low propensity to develop lymph node metastases.

Cumulative probability of prostate cancer detection in biopsy according to free/total PSA ratio in men with total PSA levels of 2.1–10.0 ng/ml at population screening

The aim of this study was to investigate the cumulative probability of prostate cancer detection according to free/total prostate-specific antigen (PSA) ratio in men with PSA levels of 2.1-10.0ng/ml and also likelihood of detecting clinically insignificant prostate cancer in population-based screening. A total of 1,277 men aged between 55 and 69years with total PSA (tPSA) levels of 2.1-10.0ng/ml screened in population screening in Kanazawa city and underwent systematic transrectal ultrasonography-guided prostate biopsy between 2000 and 2011 were enrolled. The cumulative probability of prostate cancer detection in biopsy according to age, serum tPSA, and free-to-total PSA (f/t PSA) ratio was investigated. The clinicopathological features of screening-detected prostate cancer were also investigated. Of the 1,277 subjects in the study population, 320 (25.0%) were diagnosed with prostate cancer during the observation period. The probabilities of prostate cancer detection at 3years were 64.5, 41.2, 28.5, and 14.3% for the men with f/t PSA ratio ≤0.08, 0.09-0.13, 0.14-0.22, and ≥0.23, respectively; the differences in probabilities of prostate cancer detection among men with different f/t PSA ratios were statistically significant. Among 320 patients, 84 (26.3%) had favorable clinicopathological features that made them suitable for active surveillance. The f/t PSA ratio in unfavorable cancer patients was significantly lower that that in favorable cancer patients. The present study demonstrated that the f/t PSA ratio was a strong predictor of future cancer detection and unfavorable cancerous features in prostate biopsy in men with total PSA levels of 2.1-10.0ng/ml at population screening.

Molecular profiles of screen detected vs. symptomatic breast cancer and their impact on survival: results from a clinical series.

BackgroundStage shift is widely considered a major determinant of the survival benefit conferred by breast cancer screening. However, factors and mechanisms underlying such a prognostic advantage need further clarification. We sought to compare the molecular characteristics of screen detected vs. symptomatic breast cancers and assess whether differences in tumour biology might translate into survival benefit.MethodsIn a clinical series of 448 women with operable breast cancer, the Kaplan-Meier method and the log-rank test were used to estimate the likelihood of cancer recurrence and death. The Cox proportional hazard model was used for the multivariate analyses including mode of detection, age at diagnosis, tumour size, and lymph node status. These same models were applied to subgroups defined by molecular subtypes.ResultsScreen detected breast cancers tended to show more favourable clinicopathological features and survival outcomes compared to symptomatic cancers. The luminal A subtype was more common in women with mammography detected tumours than in symptomatic patients (68.5 vs. 59.0%, p=0.04). Data analysis across categories of molecular subtypes revealed significantly longer disease free and overall survival for screen detected cancers with a luminal A subtype only (p=0.01 and 0.02, respectively). For women with a luminal A subtype, the independent prognostic role of mode of detection on recurrence was confirmed in Cox proportional hazard models (p=0.03). An independent role of modality of detection on survival was also suggested (p=0.05).ConclusionsMolecular subtypes did not substantially explain the differences in survival outcomes between screened and symptomatic patients. However, our results suggest that molecular profiles might play a role in interpreting such differences at least partially.Further studies are warranted to reinterpret the efficacy of screening programmes in the light of tumour biology.

Abstract P2-08-01: PIK3CA mutations associate with decreased Ki67 in early stage breast cancer (BC) and better outcomes in patients even among those with low Ki67 tumors.

Abstract Background: In studies of over 450 BC patients (pts) with &amp;gt; 10 years follow-up, pts with PIK3CA-mutated primary tumors had improved clinical outcome (Kalinsky et al 2009, Cizkova et al 2012). PIK3CA mutations associated with favorable clinicopathologic features: lower grade, smaller size, lymph node negativity (−), ER positivity (+), HER2−. In BC, several studies have suggested that PIK3CA mutations do not associate with PI3K/mTOR pathway activation (Loi et al 2010, Stemke-Hale et al 2008). To gain additional insight into the favorable biology imparted by a PIK3CA mutation in early stage BC and to identify predictive biomarkers, we performed immunohistochemistry (IHC) on tissue microarrays (TMA) constructed from 590 primary BCs. Methods: TMAs derived from FFPE tumors previously genotyped for PIK3CA mutations (rate of 32.5%, 192/590) were stained for ER, HER2 and AR (Kalinsky et al 2009). Here, we performed additional stains for MIB1 (Ki67 proliferation index, Ki67), p53 and markers of PI3K pathway activation (pS6, PTEN). Slides were scanned with Aperio ScanScope XT and segmented using TMALab (Aperio). Images were analyzed with the Aperio algorithm based on staining pattern (nuclear, cytoplasmic, membrane) and scored for % + and intensity. Manual review was performed for tumors with less than 3 cores or discordant results. PTEN was scored manually: 0 (no tumor staining, PTEN loss), 1 (tumor &amp;lt; stroma), or 2 (tumor ≥ stroma). P values were based on the log-rank test for comparison of Kaplan-Meier curves for disease free survival (DFS), Chi-square test for categorical variables and t-test for continuous variables. Results: On average, 66 cases (11%) were non-informative for each IHC stain, due to missing cores or insufficient tumor cells. In a binary analysis of Ki67 using a cutoff of ≥10% + cells, PIK3CA mutated tumors demonstrated significantly lower proliferation index with only 9.4% of tumors (16/170) having high Ki67 as compared to 23.6% of wild-type PIK3CA tumors (82/347) (P = .001). Ki67 was also highly associated with PIK3CA genotype when analyzed with a cutoff of ≥13.25% + cells or as a continuous variable. Importantly, DFS was significantly different when analyzed by PIK3CA genotype and Ki67 (P = .01). Among pts with low Ki67 tumors (n = 419), PIK3CA mutation associated with longer DFS (10yr 79%, CI 69–85) as compared to wild-type PIK3CA (10yr 71%, CI 64–77). When PI3K pathway interactions were analyzed, PTEN loss associated with wild-type PIK3CA (P&amp;lt;.001), although PTEN loss occurred with mutated PIK3CA in 11 hormone receptor (HR)+ tumors. In analysis of pS6 with a binary cutoff of ≥20% + cells, high pS6 associated with HER2+ (P&amp;lt;.001), HR- (P&amp;lt;.001) and high tumor grade (P = 0.003), but did not associate with PIK3CA genotype (P = .13). PIK3CA mutation was associated with lower pS6 when pS6 % + cells was analyzed as a continuous variable (P = .01). Conclusions: PIK3CA mutation associates with lower Ki67 in early stage BC. Even among pts with low Ki67 tumors, PIK3CA mutation associates with improved clinical oucome. pS6 is not increased in PIK3CA-mutated tumors which likely indicates that mTORC1 signaling is not activated as a result of PIK3CA mutation in early stage ER+ BC. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-08-01.

Clinicopathological significance of MAML2 gene split in mucoepidermoid carcinoma

CRTC1-MAML2 and CRTC3-MAML2 fusions have been associated with favorable clinicopathological features of mucoepidermoid carcinomas. However, the significance of the MAML2 gene split has not been fully clarified. In the present study, 95 mucoepidermoid carcinomas (paraffin-embedded materials) were analyzed for CRTC1-MAML2 and CRTC3-MAML2 fusions by RT-PCR and for the MAML2 gene split by FISH. Quantitative RT-PCR for the CRTC1-MAML2 transcript was performed in selected cases. MLL gene involvement, which has been reported in some leukemia cases, was examined by FISH in fusion partner-unknown cases. CRTC1-MAML2 and CRTC3-MAML2 fusions were detected in 37 and 6 cases, respectively. The MAML2 gene split was detected in 62 cases, which included all CRTC1/3-MAML2 fusion-positive cases. The level of CRTC1-MAML2 transcript expression was highly variable, and its clinicopathological impact was unclear. The MLL gene split was not detected. Mucoepidermoid carcinomas negative for CRTC1/3-MAML2 and positive for the MAML2 gene split (n = 19) showed favorable clinicopathological tumor features similar to those positive for CRTC1/3-MAML2 fusions. Compared with negative cases (n = 33), mucoepidermoid carcinomas positive for the MAML2 split (n = 62) were associated with lower patient age, a mild female predilection, a smaller tumor size, less frequent nodal metastasis, a lower clinical stage, a lower histological grade, and longer overall and disease-free survival. The MAML2 gene split emerged as an independent prognostic factor for both overall and disease-free survival in multivariate prognostic analysis. The presence of the MAML2 gene split defines a distinct mucoepidermoid carcinoma subset that is associated clinicopathologically with favorable tumor features.

CHARACTERISTICS OF PFTIENS WITH THYROID CANCER ACCOMPANIED BY BENIGN THYROID PATHOLOGY: AN ANALYSIS OF 677 CASES

Results of the study revealed the oncological process to show less favorable clinicopathologicalfeatures in patients with thyroid cancer (TC) only as compared to TC comorbid by bening thyroidneoplasms. It determines that TC accompanied by other thyroid lesions exhibited better prognosticfeatures such as low proportion of male patients, smaller size of primary tumor due to elevated numberof cases with thyroid microcarcinoma, low proportion of biologically aggressive medullary TC. Theage over 45 years was found to be only one unfavorable factor for patients with TC comorbid beningthyroid lesions.

Late recurrence of renal cell carcinoma &gt;5 years after surgery: clinicopathological characteristics and prognosis

What's known on the subject? and What does the study add? Late recurrence more than five years after the initial treatment is one of the biological behaviours specific for RCC. In our study, late recurrence was observed in 8.8% of the patients. Also, patients with late recurrence had more favorable clinicopathological features and better prognosis with long cancer-specific survival after recurrence. Age and preoperative hs-CRP levels may be independent predictive factors for late recurrence of RCC. • To evaluate the clinicopathological features and prognosis of late recurrence of renal cell carcinoma (RCC). • A total of 747 patients who had undergone curative surgery for RCC with follow-up of >5 years or recurrence within 5 years were included in the study. • The patients were stratified into four groups based on cancer-free intervals: no recurrence (no recurrence >5 years after surgery, n= 425), synchronous metastasis (n= 138), early recurrence (recurrence within 5 years, n= 143), and late recurrence (recurrence after 5 years, n= 41). • Multivariate analysis was performed to identify the clinicopathological factors affecting late recurrence and its clinical outcome. • The subgroups were significantly different in clinicopathological variables, including age, preoperative haemoglobin, platelet count, high-sensitivity C-reactive protein (hs-CRP) levels, pT stage and nuclear grade. • In multiple logistic regression analysis, age (odds ratio [OR] 1.085, 95% confidence interval [CI] 1.012-1.163, P= 0.022), and preoperative hs-CRP levels (OR 6.211, 95% CI 1.590-24.270, P= 0.009) were independent predictive factors for late recurrence. • In patients with synchronous metastasis, early recurrence and late recurrence, 5-year cancer-specific survival rates after recurrence were 27.0%, 41.1% and 73.7%, respectively (P < 0.001). • Multivariate Cox analysis indicated that cancer-free interval, as well as body mass index, initial symptoms, Fuhrman's nuclear grade, sarcomatoid differentiation, lymphovascular invasion and metastasectomy, were independent predictive factors for cancer-related death. • Late recurrence of RCC is not a rare event. • Patients with late recurrence had more favourable clinicopathological features and better prognosis with long cancer-specific survival after recurrence. • Age and preoperative hs-CRP levels may be independent predictive factors for late recurrence of RCC.

Clinicopathological significance of SOX4 expression in primary gallbladder carcinoma

AimSOX4, as a member of the SRY-related HMG-box (SOX) transcription factor family, has been demonstrated to be involved in tumorigenesis of many human malignancies; however, its role in primary gallbladder carcinoma (PGC) is still largely unknown. The aim of this study was to investigate SOX4 expression in PGC and its prognostic significance.MethodsFrom 1997 to 2006, 136 patients underwent resection for PGC. The median follow-up was 12.8 months. Immunostainings for SOX4 were performed on these archival tissues. The correlation of SOX4 expression with clinicopathological features including survival was analyzed.ResultsSOX4 was expressed in 75.0% (102/136) of PGC but not in the normal epithelium of the gallbladder. In addition, the over-expression of SOX4 was significantly associated with low histologic grade (P = 0.02), low pathologic T stage (P = 0.02), and early clinical stage (P = 0.03). The levels of SOX4 immunostainings in PGC tissues with positive nodal metastasis were also significantly lower than those without (P = 0.01). Moreover, Kaplan-Meier curves showed that SOX4 over-expression was significantly related to better overall (P = 0.008) and disease-free survival (P = 0.01). Furthermore, multivariate analyses showed that SOX4 expression was an independent risk factor for both overall (P = 0.03, hazard ratio, 3.682) and disease-free survival (P = 0.04, hazard ratio, 2.215).ConclusionOur data indicate for the first time that the over-expression of SOX4 in PGC was significantly correlated with favorable clinicopathologic features and was an independent prognostic factor for better overall and disease-free survival in patients. Therefore, SOX4 might be an auxiliary parameter for predicting malignant behavior for PGC.Virtual slidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1534825818694957.

Prognostic influence of BCL2 expression in breast cancer

Although BCL2 has occasionally been suggested as a candidate prognostic factor for breast cancer, it is still not accepted as a prognostic factor. We attempted to validate the role of BCL2 as a prognostic factor of breast cancer. Data on 7,230 primary breast cancer patients from the Seoul National University Hospital Breast Care Center were analyzed. Three current prognostic models, including the St. Gallen model, the Nottingham prognostic index (NPI) model and the TNM model, were used for analysis of the prognostic influence of BCL2. The positive BCL2 group showed more favorable features with regard to clinicopathologic parameters than the BCL2 negative group and a strong correlation was observed between BCL2 and the hormonal receptor. The positive BCL2 group showed better prognosis in overall survival and disease free survival (log-rank test, both p < 0.001), even in all subgroups, than the BCL2 negative group. BCL2 was a significant prognostic factor in both univariate (hazard ratio [HR], 0.361; 95% confidence interval (CI), 0.306-0.426; p < 0.001) and multivariate analyses (HR, 0.417; 95% CI, 0.417-0.705; p < 0.001). BCL2 had a strong influence on the established prognostic models, including the St. Gallen model, the NPI model and the TNM model. BCL2 was a powerful independent prognostic factor for breast cancer and had a strong influence on the current prognostic models. Favorable clinicopathologic features and a strong correlation with the hormonal receptor are suggested as the causes of superior survival in patients with BCL2 positive breast cancer.

Higher expression of EphA2 and ephrin-A1 is related to favorable clinicopathological features in pathological stage I non-small cell lung carcinoma

BackgroundThe overexpression of receptor tyrosine kinase EphA2 has been reported in various cancers. In non-small cell lung cancer (NSCLC), a positive correlation has been reported between high EphA2 immunohistochemical staining level and poor prognosis. However, its ligand, ephrin-A1, is supposed to act as a tumor suppressor via the kinase activity of EphA2. Thus, the biphasic roles of this system are not fully elucidated. We retrospectively evaluated the expression levels of EphA2 and ephrin-A1 in surgically treated pathological (p-) stage I NSCLC tumor samples, and their relation to clinicopathologic features or postoperative prognoses. MethodsThe levels of EphA2 and ephrin-A1 mRNA expression were quantified by real-time reverse-transcription polymerase chain reaction in tissue samples from p-stage I NSCLC patients who had undergone complete resection in our facility (n=195). They were divided into two (EphA2/ephrin-A1-Low and -High) groups based on the median expression level, and their respective clinicopathologic features and prognoses were analyzed. Furthermore, samples were stained immunohistochemically and classified into four groups according to their staining levels, and their prognoses analyzed. ResultsMarked demographic differences were found between EphA2/ephrin-A1-Low and -High groups. Both EphA2-High and ephrin-A1-High groups had more females, no smoking history, adenocarcinoma histology, well-differentiated carcinomas, p-stage IA patients, and patients with EGFR gene mutations. Five-year overall survival rates of the EphA2-Low and the EphA2-High patient groups were 68.9% and 86.1%, respectively (P=0.017), and five-year disease-free survival rates were 69.9% and 83.2%, respectively (P=0.035). There were no statistical differences between ephrin-A1-Low and ephrin-A1-High groups concerning postoperative survival. Although showing smaller differences, the findings from the immunohistochemical analyses supported the above results. ConclusionsHigher expression of EphA2 and ephrin-A1 was more related to the female sex, reduced smoking status, adenocarcinoma, well differentiated carcinomas, p-stage IA, and EGFR gene mutations. Higher EphA2 mRNA expression in p-stage I NSCLC patients was positively related to improved prognoses. These results may reflect a tumor suppressive role for the EphA2/ephrin-A1 system in a population of patients restricted to p-stage I NSCLC.

Abstract P3-10-03: The Implication of Androgen Receptor Expression in Breast Cancers

Abstract Purpose: To evaluate the implication of androgen receptor (AR) expression in patients with breast cancer. Methods: Immunohistochemically AR expression was investigated in 931 invasive breast cancers prepared in tissue microarray blocks that were collected in between November 1999 and August 2005. The clinicopathological features, disease-free (DFS) and overall survival (OS) were analyzed using chi-square test, Kaplan-Meier methods, and Cox's models. Tumors with ≥10% nuclear-stained cells were considered to be positive for AR, estrogen receptor (ER), and progesterone receptor (PR) expression and 3+ with immunohistochemical staing or FISH + were considered positive for HER2 expression. Results: The positive expression of AR, ER, PR, and HER2 was 58.1%, 72.2%, 61.4%, and 24.7% of all patients, respectively. AR expression was significantly associated with an older age at diagnosis (p=0.007), smaller tumor size (P&amp;lt;0.001), well differentiation (P&amp;lt;0.001), higher hormone receptors expression (P&amp;lt;0.001), non-triple-negative breast cancers (non-TNBC) (P&amp;lt;0.001), and lower proliferation index (P&amp;lt;0.001). In ER-negative cancers, AR expression was associated with well differentiation, lower Ki-67 index, and distinctively HER2 overexpression. With median follow-up duration of 72.7 months, AR was significantly associated with better DFS (p=0.023) and OS (p=0.048) and the association was same in ER-positive cancers but was disappeared in ER-negative cancers. In Cox's models, AR expression was an independent prognostic factor for DFS (hazard ratio, 0.654; p=0.049), but it did not reach a statistical significance for OS (hazard ratio, 0.647; p=0.119) even in ER-positive tumors. Interestingly, tumors with molecular apocrine features (ER-negative, AR-positive, and HER2-positive) showed a trend of poorer outcome with AR expression, but there was no impact of AR expression on the survival in 156 patients with TNBC. Conclusions: AR expression is significantly associated with favorable clinicopathological features and better outcomes in ER-positive cancers but tumors with molecular apocrine features showed a trend of poorer outcome with AR expression. These results suggest that AR expression could be an additional marker for endocrine responsiveness in ER-positive tumors and a target molecule for the molecular apocrine tumors. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-10-03.

Mutant PIK3CA Is Detected in Both Pre-Invasive and Recurrent Breast Cancer.

Abstract BackgroundWe identified somatic PIK3CA mutations in 32.5% of 590 primary invasive breast cancers (BC) (manuscript in press: Clinical Cancer Research). Detected by massARRAY genotyping, PIK3CA mutations significantly associate with favorable clinicopathologic features and improved clinical outcome, including overall and breast cancer-specific survival. Given the strong association between PIK3CA mutations and hormone receptor (HR) positivity, one hypothesis is that PIK3CA mutations 'drive' HR positive BC and will be detected in pre-invasive breast tumors. As PIK3CA mutations offer a protective effect in BC, it has not been determined whether PIK3CA mutations are selected for in disease progression or whether additional collaborating mutations are required.MethodsTo determine the concordance of PIK3CA mutations and assess the acquisition of additional oncogene mutations, available matched tissue samples, from the previous database, were procured and underwent massARRAY genotyping (n = 83). Two mm cores were macro-dissected from matched formalin-fixed, paraffin embedded tissue, including normal breast tissue, benign lymph nodes (LN), ductal carcinoma-in-situ (DCIS), regional LN metastases (mets), and distant mets. MassARRAY (Sequenom) genotyping was performed on native DNA to identify rare and hotspot PIK3CA mutations, as well as AKT1 (E17K), RAS, and RET mutations.ResultsConcordance of PIK3CA mutations is noted between primary BC and DCIS, except for one rare PIK3CA mutation (Q546R) not detected in DCIS (4/5). No PIK3CA mutations are detected in normal breast tissue (0/8) or benign LN (0/6). Rare and hotspot PIK3CA mutations in primary BC are detected in 87.5% (7/8) matched regional LN met and 80 % (4/5) distant mets. For the patient with the disconcordant regional LN met, the rare PIK3CA mutation (H1047L) identified in primary BC is also present in the distant met site. Notably, for the single disconcordant PIK3CA met site, a rare PIK3CA mutation (E545A) is detected in a primary tumor, regional LN met, and bone met and is absent in a second bone met, in which a KRAS (G12C) mutation has been identified. One concomitant hotspot PIK3CA (E542K)/KRAS (G12C) mutation is present in both primary BC and paired DCIS. Complete concordance of AKT1 (E17K) mutations has been identified between in DCIS, primary BC, regional LN met, and distant mets (n=4).ConclusionsWe recently defined the positive prognostic significance of PIK3CA mutations in breast cancer. PIK3CA and AKT1 (E17K) mutations are early events in breast cancer, occurring in pre-invasive tumors. Despite the protective effect of PIK3CA mutations on clinical outcome, they persist and are selected for in disease progression, as they are detected in regional LN and distant mets. Complete concordance is identified between hotspot PIK3CA mutated primary BC and matched tumors samples, suggesting that PIK3CA mutations with higher oncogenic potency are maintained in tumor progression. These findings support that targeting the PI3K pathway may assist in tailoring therapy to appropriate patient populations. We are expanding matched tissue collection from a separate patient cohort to further assess genomic and functional genomic change with disease progression. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5164.

Clinicopathological significance of the CRTC3–MAML2 fusion transcript in mucoepidermoid carcinoma

Mucoepidermoid carcinoma is the most common primary malignancy of the salivary gland. We and others showed that CRTC1–MAML2 gene fusion was associated with favorable clinicopathological tumor features. Recently, a novel gene fusion, CRTC3–MAML2, was reported as a rare gene alteration in a case of mucoepidermoid carcinoma. However, its frequency and clinicopathological significance remains unclear. In all, 101 cases of mucoepidermoid carcinoma and 89 cases of non-mucoepidermoid carcinoma of the salivary gland were analyzed, and RNA was extracted from formalin-fixed, paraffin-embedded specimens. In the CRTC family, there have been three genes, CRTC1, CRTC2, and CRTC3. We developed reverse transcription-polymerase chain reaction (RT-PCR) assays for CRTC1–MAML2, CRTC2–MAML2, and CRTC3–MAML2 fusions. Clinicopathological data of the patients were obtained from their clinical records. Of 101 cases of mucoepidermoid carcinoma, 34 (34%) and 6 (6%) were positive for CRTC1–MAML2 and CRTC3–MAML2 fusion transcripts. However, in the 89 cases of non-mucoepidermoid carcinoma, neither transcript was noted. In the former cases, CRTC1–MAML2 and CRTC3–MAML2 fusions were mutually exclusive. The other fusion, CRTC2–MAML2, was not detected. We confirmed that the clinicopathological features of CRTC1–MAML2-positive mucoepidermoid carcinomas indicated an indolent course. CRTC3–MAML2-positive mucoepidermoid carcinomas also had clinicopathologically favorable features; all cases showed a less advanced clinical stage, negative nodal metastasis, no high-grade tumor histology, and no recurrence or tumor-related death after surgical resection of the tumor. It is interesting to note that patients with CRTC3–MAML2-positive tumors (mean 36 years of age) were significantly younger that those with the CRTC1–MAML2 fusion (55 years) and those with fusion-negative tumors (58 years). In conclusion, CRTC3–MAML2 fusion, which is mutually exclusive with CRTC1–MAML2 fusion and specific to mucoepidermoid carcinoma, may be detected more frequently than previously expected. Mucoepidermoid carcinomas possessing CRTC3–MAML2 fusion may be associated with favorable clinicopathological features and patients may be younger than those with CRTC1–MAML2 fusion or those with no detectable gene fusion.

The prognosis of gastric cardia cancer after R0 resection

Background The aim of this study was to evaluate the prognosis of gastric cardia cancers in comparison with other gastric cancers. Methods The medical records of 251 patients with gastric cardia cancers and 6568 patients with other gastric cancers who underwent R0 resection were reviewed. Clinicopathologic characteristics and survival were analyzed. Results Gastric cardia cancer was associated with more advanced staging and less favorable clinicopathologic features at diagnosis compared with other gastric cancers. The overall 5-year survival rates were 79.7% and 84.6% in patients with cardia cancer and other cancers, respectively. There were no significant differences in survival curves between the groups at any stage. Lymph node metastasis was an independent prognostic factor for disease-free survival. The length of the proximal margin was not associated with locoregional tumor recurrence. Conclusions Although patients with gastric cardia cancers are diagnosed at an advanced stage, the long-term survival rates are similar to those with other gastric cancers. If curative resection with negative resection margin can be achieved, pN category is the only prognostic factor for survival.

Phosphoinositide 3-Kinase Mutations in Breast Cancer: A “Good” Activating Mutation?

Gain-of-function mutations in the catalytic subunit of phosphoinositide-3-kinase (PI3KCA) occur frequently in breast cancer. Kalinsky and colleagues show that PI3KCA mutations are associated with favorable clinicopathologic features and better clinical outcome, including survival. These findings will have to be considered in the design and interpretation of clinical trials with inhibitors of the PI3K pathway.

Prognostic Significance of p27 Kip1, Ki-67, and CRTC1- MAML2 Fusion Transcript in Mucoepidermoid Carcinoma: A Molecular and Clinicopathologic Study of 101 Cases

Mucoepidermoid carcinoma (MEC) is the most frequently detected primary malignancy of the salivary gland and is characterized by a marked variation in prognosis. In the present study, we investigated the prognostic significance of p27Kip1, Ki-67, and CRTC1 (also called MECT1, TORC1, and WAMTP1)-MAML2 fusion in MEC. MEC cases (n = 101) were examined for p27Kip1 and Ki-67 expression using immunohistochemistry and for CRTC1-MAML2 fusion transcript using reverse transcriptase-polymerase chain reaction. p27Kip1, Ki-67, and the CRTC1-MAML2 fusion transcript were expressed in 71, 31, and 34 of the 101 cases, respectively. p27Kip1 and CRTC1-MAML2 fusion were associated with favorable clinicopathologic tumor features and Ki-67 with aggressive clinicopathologic features. Multivariate survival analyses were performed that included the following 10 clinicopathologic factors: age, gender, tumor site, tumor size, nodal metastasis, clinical stage, histologic grade, p27 expression, Ki-67 expression, and CRTC1-MAML2 fusion. For disease-free survival, only p27Kip1 expression was significant as an independent prognostic factor. For overall survival, p27Kip1 expression, CRTC1-MAML2 fusion, and tumor size were significant. In each analysis, p27Kip1 and CRTC1-MAML2 fusion were independent of the clinical stage. Ki-67 expression was not selected in either multivariate analysis. p27Kip1 and CRTC1-MAML2 fusion were associated with favorable clinicopathologic tumor features, and both were useful in predicting the overall survival of patients with MEC. For disease-free survival, p27Kip1 might be the most useful prognostic factor. In contrast, Ki-67 might not be a very powerful prognostic indicator for either survival point.