Characterization of the immunological microenvironment of tumour buds and its impact on prognosis in mismatch repair-proficient and -deficient colorectal cancers

Zlobec I, Minoo P, Terracciano L, Baker K & Lugli A
(2011) Histopathology 59 , 482–495
Characterization of the immunological microenvironment of tumour buds and its impact on prognosis in mismatch repair‐proficient and ‐deficient colorectal cancers Aims: Tumour budding in colorectal cancer is established as a poor prognostic factor. The inverse correlation of tumour buds with peritumoural lymphocytic inflammation suggests an interaction with specific immune responses. The aims of this study were to characterize the immunological microenvironment of tumour buds and its impact on prognosis in mismatch repair (MMR)‐proficient and ‐deficient colorectal cancers. Methods and results: A total of 297 colorectal cancers were double‐immunostained for CK22 plus one of the following: CD138, CD16, CD20, CD21, CD56, CD68, CD8, forkhead box P2 (FoxP3), granzyme B, mast cell tryptase, CD3 or T cell intracellular antigen‐1 (TIA)‐1. Tumour buds and immune cells within the region of densest budding were evaluated [×40 high‐power field (HPF)] simultaneously. In both MMR‐proficient and ‐deficient cancers, CD8⁺, FoxP3⁺ and CD68⁺ cells were observed most frequently (>40 cells/HPF) and were independent prognostic factors. A combined prognostic score of tumour budding and CD8⁺, FoxP3⁺ and CD68⁺ distinctly identified patients with low‐, moderate‐ or high‐risk colorectal cancers with 5‐year survival rates of 75.2% [confidence interval 95% (CI): 66–83], 56.3% (95% CI: 43–68) and 25.2% (95% CI: 14–38), respectively, in MMR‐proficient and ‐deficient cancers. Conclusion: The combined assessment of tumour budding with CD8, FoxP3 and CD68 lymphocytes could represent a basis for a prognostic score similar to the Bloom Richardson grade (BRE) and Gleason scores for breast and prostatic cancers.     

Digital image analysis of HER2 immunohistochemistry in gastric‐ and oesophageal adenocarcinoma: a validation study on biopsies and surgical specimens

Aims

To test the validity of diagnostics incorporating digital image analysis (DIA) for human epidermal growth factor 2 (HER2) immunohistochemistry (IHC) in gastro‐oesophageal adenocarcinomas, as an alternative to current standard diagnostics using manual scoring.

Methods and results

We included 319 consecutive gastro‐oesophageal adenocarcinomas (232 biopsies and 87 surgical specimens). DIA was applied to determine HER2 IHC classification, using both standard breast cancer (BC) and modified gastro‐oesophageal cancer (GEC) cut‐offs. Consensus manual scores were established by four independent observers. Chromogenic in‐situ hybridization (CISH) was performed on all 2+ cases by manual scoring, DIA or both. HER2 status was considered positive in 3+ and CISH‐positive 2+ cases. Overall agreement between DIA and consensus manual scores was 76.5% (weighted κ = 0.66, BC cut‐offs) and 85.6% (weighted κ = 0.80, GEC cut‐offs). Agreement was similar for biopsies and surgical specimens. All disagreement occurred in the manual IHC equivocal cases. DIA resulted in a reduction of 2+ cases: 75.8% with BC cut‐offs and 46.5% with GEC cut‐offs. HER2 status was positive in 48 cases (15%) with standard diagnostics and DIA using GEC cut‐offs, and 46 cases (14.4%) using BC cut‐offs (all with CISH in 2+ cases). Considering standard diagnostics as a reference, DIA showed 93.8% sensitivity and 99.6% specificity (BC cut‐offs) or 97.9% sensitivity and 99.6% specificity (GEC cut‐offs).

Conclusions

DIA is a reliable and feasible alternative to manual HER2 IHC scoring in gastro‐oesophageal adenocarcinoma, both in biopsies and surgical specimens, leading to a reduction of 2+ cases for which subsequent ISH testing is required.     

Cytoplasmic β‐catenin accumulation is a good prognostic marker in upper and lower gastrointestinal adenocarcinomas

Norwood M G A, Bailey N, Nanji M, Gillies R S, Nicholson A, Ubhi S, Darnton J J, Steyn R S, Womack C, Hughes A, Hemingway D, Harrison R, Waters R & Jankowski J A
(2010) Histopathology 57, 101–111
Cytoplasmic β‐catenin accumulation is a good prognostic marker in upper and lower gastrointestinal adenocarcinomas Aims: β‐Catenin is an important molecule in cancer biology. Membranous β‐catenin enhances cellular differentiation and inhibits invasion by its action on E‐cadherin. The aim was to ascertain whether the cellular expression of these molecules in colorectal and oesophageal cancer specimens is associated with survival in patients with gastrointestinal cancer. Methods and results: Tumour samples from 149 patients undergoing resection for colorectal adenocarcinoma and 147 patients undergoing resection for oesophageal adenocarcinoma were retrospectively analysed using immunohistochemical techniques to assess β‐catenin expression. Increasing β‐catenin expression in the cytoplasm was associated with improved survival for colorectal cancer cases on both univariate (P = 0.003) and multivariate (P = 0.01) analysis. In addition, increased expression in the most recent cohort of oesophageal adenocarcinoma patients was associated with improved TNM staging (P = 0.007). Membrane expression was weakly associated with survival in colorectal cancer on univariate analysis (P = 0.09), but not on multivariate analysis (P = 0.21). Complete absence of β‐catenin expression at all three sites was associated with reduced 5‐year survival in colorectal cancer. Conclusions: This is one of the largest prognostic studies of β‐catenin in gastrointestinal adenocarcinoma. It shows that low levels of cytoplasmic β‐catenin expression are associated with reduced survival in patients with colorectal cancer as well as worse TNM staging in oesophageal adenocarcinoma (a recognized surrogate end‐point for survival). We believe this is the first time that this has been reported. This finding should be tested prospectively in oncological trials to validate whether the presence of cytoplasmic β‐catenin could be used as a prognostic marker for less aggressive disease.     

HER2 status in gastro-oesophageal adenocarcinomas assessed by two rabbit monoclonal antibodies (SP3 and 4B5) and two in situ hybridization methods (FISH and SISH)

Boers J E, Meeuwissen H & Methorst N
(2011) Histopathology 58 , 383–394
HER2 status in gastro‐oesophageal adenocarcinomas assessed by two rabbit monoclonal antibodies (SP3 and 4B5) and two in situ hybridization methods (FISH and SISH) Aims: HER2 gene amplification has been detected in 10–20% of gastric adenocarcinomas. In view of the recently demonstrated clinical benefit of the anti‐human epidermal growth factor receptor 2 (HER2) drug trastuzumab in the treatment of advanced gastric cancer, reliable HER2 testing is of key importance. The aim of this study was to examine HER2 status in gastro‐oesophageal adenocarcinomas comparing SP3 and 4B5 immunohistochemistry (IHC) with dual probe HER2 [fluorescence in situ hybridization (FISH) and silver in situ hybridization (SISH)]. Methods and results: IHC and SISH were carried out on biopsy specimens of 146 patients with adenocarcinomas of the oesophagus and stomach. All SP3‐IHC‐positive cases and 91% of 4B5‐IHC‐positive cases were amplified. Sensitivity of SP3‐IHC‐positivity and 4B5‐IHC‐positivity for amplification was 77% and 96%, respectively. Results of FISH performed in 42 cases were identical to SISH. Amplification was heterogeneous in 73% of the adenocarcinomas; 24% of the oesophago‐gastric carcinomas and 7% of distal stomach tumours were amplified. Conclusions: HER2‐positivity is present in a significant proportion of oesophago‐gastric adenocarcinomas (24%), but at a lower rate in the distal stomach (7%). Sensitivity for amplification is higher with 4B5 IHC than with SP3. FISH and SISH yield identical results, but assessment is much easier with SISH. Our findings provide important guidance for HER2‐testing in gastro‐oesophageal adenocarcinomas for patients in whom anti‐HER2 treatment is considered.     

Tumour budding and the expression of cancer stem cell marker aldehyde dehydrogenase 1 in nasopharyngeal carcinoma

Luo W‐R, Gao F, Li S‐Y & Yao K‐T
(2012) Histopathology
Tumour budding and the expression of cancer stem cell marker aldehyde dehydrogenase 1 in nasopharyngeal carcinoma Aims: To detect the prognostic significance of tumour budding and its expression of aldehyde dehydrogenase 1 (ALDH1) in nasopharyngeal carcinoma (NPC). Methods and results: Tumour budding was investigated in 105 patients with NPC by immunohistochemistry for pan‐cytokeratin (AE1/AE3). The intensity of budding correlated strongly with T classification (P = 0.008), lymphatic invasion (P < 0.001), vascular invasion (P = 0.029), lymph node metastasis (P < 0.001), and clinical stage (P = 0.010). Univariate analysis revealed that patients with high budding grade had poorer survival than those with low grade (P = 0.002). Multivariate analysis showed that tumour budding was an independent predictor of survival (P = 0.001). Furthermore, budding cells showed high‐level expression of the cancer stem cell (CSC) marker ALDH1. Budding cells with high‐level ALDH1 expression contributed to several aggressive behaviours and poor survival (P = 0.000). Conclusions: We describe, for the first time, the presence of tumour budding and its correlation with aggressive tumour behaviour and poor patient survival in NPC. The degree of tumour budding could be a valuable predictive factor in NPC. In addition, we show, also for the first time, that budding cells in NPC might possess the invasive and metastatic properties of CSCs.     

The impact of CpG island methylator phenotype and microsatellite instability on tumour budding in colorectal cancer

Zlobec I, Bihl M P, Foerster A, Rufle A & Lugli A
(2012) Histopathology  61, 777–787 The impact of CpG island methylator phenotype and microsatellite instability on tumour budding in colorectal cancer Aims: In colorectal cancer, tumour budding, a process likened to epithelial mesenchymal transition, is an adverse prognostic factor which is rarely found in tumours with high‐level microsatellite instability (MSI‐H). Cases with MSI‐H or high‐level CpG island methylator phenotype (CIMP‐H) have similar histomorphological features, yet seemingly opposite prognosis. We hypothesized that tumour budding is related to CIMP, thus partially explaining this prognostic difference. Methods and results: MSI, KRAS, BRAF, CIMP and 0⁶‐methylguanine‐DNA methyltransferase (MGMT) were investigated in tissues from 127 colorectal cancer patients. Tumour budding was scored using pan‐cytokeratin‐stained whole tissue sections within the densest area of buds (×40). Tumour budding was not associated with KRAS, BRAF, MGMT or CIMP, but was correlated inversely with MSI‐H (P = 0.0049). Multivariate survival time analysis revealed that tumour budding was independent of all five molecular features and was predicted by MSI status [odds ratio (OR): 4.29, 95% confidence interval (CI) 1.5–12.1; P = 0.006)], but not CIMP (OR: 0.81, 95% CI 0.3–2.5; P = 0.714). Conclusions: These findings underline that MSI, rather than CIMP, plays a role in conferring a tumour budding phenotype. Budding retains its unfavourable prognostic effect independently of these five molecular features. Continued efforts to standardize the assessment of tumour budding are necessary to integrate this feature into daily diagnostic routine.     

Comparison of gastro-oesophageal junction carcinomas in Chinese versus American patients

Huang Q, Fan X, Agoston A T, Feng A, Yu H, Lauwers G, Zhang L & Odze R D
(2011) Histopathology 59 , 188–197 Comparison of gastro‐oesophageal junction carcinomas in Chinese versus American patients Aims: To compare the clinical and pathological features of gastro‐oesophageal junction (GEJ) carcinomas in Chinese and American patients. Methods and results: Eighty consecutive patients with a GEJ carcinoma (43 from mainland China, and 37 from the USA) were evaluated for association with Barrett oesophagus (BO), chronic Helicobacter pylori gastritis, intestinal metaplasia, and outcome. GEJ carcinomas were defined as tumours that were located within 20 mm of, and crossed, the GEJ. Overall, GEJ carcinomas from Chinese patients revealed significantly more frequent location in the proximal stomach, higher pathological stage, larger size, younger patient age, and association with chronic H. pylori gastritis. In contrast, GEJ cancers from American patients showed a strong association with distal oesophageal location, BO, and associated intestinal metaplasia and dysplasia. Pathologically, GEJ carcinomas from American patients were predominantly adenocarcinomas, whereas Chinese patients showed a higher proportion of mucinous, adenosquamous, acinar or neuroendocrine tumours. Overall, 3‐ and 5‐year survival rates were statistically similar between both patient groups, but upon multivariate analysis, Chinese patients showed statistically better survival rates for stage III tumours. Conclusions: Most GEJ carcinomas in patients from China represent proximal gastric cancers associated with chronic H. pylori gastritis, and BO‐associated carcinomas are rare among this patient population.     

CD117 expression in operable oesophageal squamous cell carcinomas predicts worse clinical outcome

Aims

To investigate the aberrant expression of CD117 in oesophageal squamous cell carcinoma (SCC) and its prognostic significance.

Methods and results

Immunohistochemical staining for CD117 was performed on tissue microarray and routine tissue sections from 157 oesophageal SCC patients and 10 normal oesophageal epithelia adjacent to tumour. The positive rate of CD117 expression was 29.9% in oesophageal SCC tissues, whereas no CD117 expression was detected in the 10 normal oesophageal epithelia. CD117 expression was significantly associated with T stage (P < 0.001), distant metastasis (P = 0.015), lymph node metastasis (P = 0.019), and clinical stage (P = 0.021). Progression‐free survival in the patients with CD117‐positive tumours was shorter than that in the patients with CD117‐negative tumours (P = 0.010). In univariate analyses, CD117 expression was the most significant factor for overall survival of oesophageal SCC patients (P < 0.001), followed by lymph node metastasis (P = 0.001), T stage (P = 0.002), clinical stage (P = 0.006), distant metastasis (P = 0.020), and histological grade (P = 0.027). Multivariate analyses verified that CD117 expression was an independent prognostic marker for oesophageal SCC patients (P = 0.002). In addition, CD117 expression predicted poorer survival in patients without distant metastases.

Conclusions

CD117 expression in operable oesophageal SCC may be a valuable prognostic marker, and detection of its expression in clinical samples may be useful in defining a subclass of oesophageal SCCs with extremely poor clinical outcome, which may require a specially targeted treatment modality.     

Expression of key hypoxia sensing prolyl‐hydroxylases PHD1, ‐2 and ‐3 in pancreaticobiliary cancer

Gossage L, Zaitoun A, Fareed K R, Turley H, Aloysius M, Lobo D N, Harris A L & Madhusudan S
(2010) Histopathology 56, 908–920
Expression of key hypoxia sensing prolyl‐hydroxylases PHD1, ‐2 and ‐3 in pancreaticobiliary cancer Aims: Tumour hypoxia is associated with an aggressive phenotype and resistance to chemotherapy and radiotherapy. The aim was to investigate whether key hypoxia sensing prolyl hydroxylases PHD1, PHD2 and PHD3 are dysregulated in pancreaticobiliary cancers, and to evaluate their potential clinical significance. Methods and results: Formalin‐fixed human pancreatic tissue from 120 consecutive patients undergoing pancreatic resections between June 2001 and June 2006 was constructed into tissue microarrays. Expression of PHD1, PHD2 and PHD3 was analysed using immunohistochemistry and correlated with clinicopathological variables and disease‐specific overall survival. PHD1, PHD2 and PHD3 were significantly overexpressed in pancreaticobiliary tumours compared with normal pancreatic ductal tissues (P = 0.03, P < 0.0001 and P < 0.0001, respectively). PHD3 expression in tumour tissue was associated with a trend towards worse overall disease‐specific survival in ampullary adenocarcinomas (P = 0.035) and pancreatic adenocarcinomas (P = 0.084). Absence of PHD1 expression was significantly associated with perineural invasion in pancreatic adenocarcinomas (P = 0.02) and a trend towards significance was also seen for absence of PHD2 expression in pancreatic adenocarcinomas (P = 0.04). Conclusions: Our results provide the first clinical evidence that PHD1, PHD2 and PHD3 may be involved in pancreaticobiliary tumorigenesis.     

Comparison of histological parameters for the diagnosis of eosinophilic oesophagitis versus gastro‐oesophageal reflux disease on oesophageal biopsy material

Aims: Eosinophil infiltration of the oesophageal epithelium is the cardinal pathomorphological finding in eosinophilic oesophagitis (EO), but gastro‐oesophageal reflux disease (GORD) is also associated with increased eosinophils. The aim was to compare histological parameters for the diagnosis of EO versus GORD on routinely taken biopsy specimens. Methods and results: One hundred and five routine biopsy specimens with EO (n = 62), GORD (n = 24) and probable EO (n = 19) from 74 patients (52 men, 22 women; mean age 43.7 years) were analysed for numbers of eosinophils, mast cells, degranulation and qualitative changes of oesophageal epithelium using immunohistochemistry with monoclonal antibodies against eosinophil peroxidase and eosinophil major basic protein and mast cell tryptase. Eosinophil infiltration was significantly higher in EO than in GORD both on haematoxylin and eosin staining (54.8 versus 9.1; P < 0.05) and immunohistochemistry (77.5 versus 24.7; P < 0.05). Eosinophil degranulation was significantly more intense in EO than in GORD (1.16 versus 0.41; P < 0.05). Furthermore, eosinophilia‐codependent secondary qualitative changes of squamous epithelium in EO were generally more extensive than those in GORD. Conclusions: Histological differential diagnosis of EO and GORD should be based on eosinophil counts, secondary morphological changes of eosinophils and oesophageal squamous epithelium, especially in cases suspicious of EO.     

Stage II colonic adenocarcinoma: a detailed study of pT4N0 with emphasis on peritoneal involvement and the role of tumour budding

Canney A L, Kevans D, Wang L M, Hyland J M P, Mulcahy H E, O’Donoghue D P, O’Sullivan J, Geraghty R & Sheahan K
(2012) Histopathology  61, 488–496 Stage II colonic adenocarcinoma: a detailed study of pT4N0 with emphasis on peritoneal involvement and the role of tumour budding Aims: Evaluation of peritoneal involvement in colonic cancer (CC) can be difficult. We studied pT4N0 cancers and their association with pathological prognostic markers, including tumour budding. Method and results: Tumours were classified as (i) at the peritoneal surface or free in the peritoneal cavity (pT4a subgroup n = 44); (ii) directly invading adjacent organ (pT4b subgroup n = 8); or (iii) showing inflammatory involvement of the peritoneum (pT4I subgroup n = 25). A published pT3N0 cohort was used to compare Stage II subgroups. Standard pathological markers including tumour budding were assessed. Elastin staining was performed in the pT4I subgroup. Seventy‐seven Stage II CCs met inclusion criteria. There was no significant difference in survival across subgroups. pT4b tumours were larger than pT4a tumours (P < 0.001). Over‐represented features in pT4a versus pT4b tumours were tumour budding (P = 0.02) and infiltrative margin (P = 0.02). Tumour budding did not predict survival. Using multivariate analysis, neural invasion was the only parameter predictive of survival (hazard ratio = 2.8; 95% CI 1.2–6.4; P = 0.02). Conclusion: Stage II pT4I CCs have a similar outcome to T4a/b tumours. Elastin staining is useful in defining this group. Tumour budding may facilitate peritoneal invasion in pT4a tumours, but does not predict outcome in pT4N0 disease. Only neural invasion independently predicted poor outcome.     

Increased Toll-like receptor 9 expression indicates adverse prognosis in oesophageal adenocarcinoma

Kauppila J H, Takala H, Selander K S, Lehenkari P P, Saarnio J & Karttunen T J
(2011) Histopathology 59 , 643–649 Increased Toll‐like receptor 9 expression indicates adverse prognosis in oesophageal adenocarcinoma Aims: Toll‐like receptor 9 (TLR‐9) is a cellular DNA receptor that has been linked previously to invasion in various cancers. The aim of this study was to investigate TLR‐9 expression and its possible association with prognosis in oesophageal adenocarcinoma. Methods and results: Immunohistochemical TLR‐9 expression was graded in clinical specimens (n = 76) of oesophageal adenocarcinoma. The TLR‐9 immunostaining intensity was compared with tumour grade, stage and indicators of proliferation, apoptosis and tumour vascular supply. High TLR‐9 expression correlated with advanced tumour stage, tumour unresectability, poor differentiation and high proliferation. Strong immunoreactivity of TLR‐9 also indicated poor overall survival. Conclusions: High TLR‐9 expression is associated with poor differentiation, a high proliferation rate and disseminated disease. Accordingly, increased TLR‐9 expression may contribute to the growth and metastatic properties of oesophageal adenocarcinoma.     

Systematic assessment of protein phenotypes characterizing high‐grade tumour budding in mismatch repair‐proficient colorectal cancer

Karamitopoulou E, Lugli A, Panayiotides I, Karakitsos P, Peros G, Rallis G, Patsouris E S, Terracciano L & Zlobec I (2010) Histopathology 57 , 233–243
Systematic assessment of protein phenotypes characterizing high‐grade tumour budding in mismatch repair‐proficient colorectal cancer Aims: A tumour bud is defined as a single tumour cell or tumour cell cluster of up to five cells at the invasive tumour front. Significant differences in survival have been detected in colorectal cancer patients with low‐ compared to high‐grade budding. The aim of this study was to identify potential multi‐marker phenotypes characterizing low‐ and high‐grade budding in mismatch repair (MMR)‐proficient colorectal cancer. Methods and results: Established and promising prognostic proteins such as epidermal growth factor receptor (EGFR), pERK, RHAMM, RKIP, β‐catenin, E‐cadherin, pAKT, p16, p21, Ki67, Bcl‐2, vascular endothelial growth factor (VEGF), apoptotic protease activating factor‐1 (APAF‐1), MUC1, EphB2, matrix metalloproteinase 7, pSMAD2, CDX2, laminin5γ2 and MST1 were analysed on 208 MMR‐proficient colorectal cancers with complete clinicopathological data. The most accurate markers for predicting high‐grade budding (more than six tumour buds) were EphB2 (P < 0.001), Bcl‐2 (P < 0.001), RKIP (P < 0.001), E‐cadherin (P = 0.004), laminin5γ2 (P = 0.004) and APAF‐1 (P = 0.005). On multivariable analysis, only loss of Bcl‐2 (P < 0.001) and EphB2 (P < 0.001) were independent predictors of high‐grade budding. Bcl‐2?/EphB2? tumours were more frequently poorly differentiated (P < 0.001), of advanced pT stage (P = 0.002), lymph node positive (P = 0.023), presented vascular (P = 0.053) and lymphatic invasion (P = 0.005) and had a negative impact on patient survival (P = 0.012). Conclusions: The multi‐marker phenotype EphB2?/Bcl‐2? is an independent predictor of high‐grade budding and implies increased aggressive behaviour in MMR‐proficient colorectal cancer.     

Prognostic value of pathological lymph node status and primary tumour regression grading following neoadjuvant chemotherapy – results from the MRC OE02 oesophageal cancer trial

Aims

Neoadjuvant chemotherapy (NAC) remains an important therapeutic option for advanced oesophageal cancer (OC). Pathological tumour regression grade (TRG) may offer additional information by directing adjuvant treatment and/or follow‐up but its clinical value remains unclear. We analysed the prognostic value of TRG and associated pathological factors in OC patients enrolled in the Medical Research Council (MRC) OE02 trial.

Methods and results

Histopathology was reviewed in 497 resections from OE02 trial participants randomised to surgery (S group; n = 244) or NAC followed by surgery [chemotherapy plus surgery (CS) group; n = 253]. The association between TRG groups [responders (TRG1–3) versus non‐responders (TRG4–5)], pathological lymph node (LN) status and overall survival (OS) was analysed. One hundred and ninety‐five of 253 (77%) CS patients were classified as ‘non‐responders’, with a significantly higher mortality risk compared to responders [hazard ratio (HR) = 1.53, 95% confidence interval (CI) = 1.05–2.24, P = 0.026]. OS was significantly better in patients without LN metastases irrespective of TRG [non‐responders HR = 1.87, 95% CI = 1.33–2.63, P < 0.001 versus responders HR = 2.21, 95% CI = 1.11–4.10, P = 0.024]. In multivariate analyses, LN status was the only independent factor predictive of OS in CS patients (HR = 1.93, 95% CI = 1.42–2.62, P < 0.001). Exploratory subgroup analyses excluding radiotherapy‐exposed patients (n = 48) showed similar prognostic outcomes.

Conclusion

Lymph node status post‐NAC is the most important prognostic factor in patients with resectable oesophageal cancer, irrespective of TRG. Potential clinical implications, e.g. adjuvant treatment or intensified follow‐up, reinforce the importance of LN dissection for staging and prognostication.     

Aldehyde dehydrogenase 1 expression is a predictor of poor prognosis in node-positive breast cancers: a long-term follow-up study

Yoshioka T, Umekita Y, Ohi Y, Souda M, Sagara Y, Sagara Y, Sagara Y, Rai Y & Tanimoto A
(2011) Histopathology 58 , 608–616
Aldehyde dehydrogenase 1 expression is a predictor of poor prognosis in node‐positive breast cancers: a long‐term follow‐up study Aims: Aldehyde dehydrogenase 1 (ALDH1) has been identified as a reliable marker of breast cancer stem cells. The aim was to determine the prognostic significance of ALDH1 expression in a long‐term follow‐up study. Methods and results: Immunohistochemical analyses were performed on 257 invasive ductal carcinomas (IDCs), 109 matched lymph node metastases and 190 ductal carcinomas in situ (DCISs), using paraffin‐embedded sections. ALDH1 expression was found in 26% of IDCs, and correlated with larger tumour size (P = 0.007), high histological grade (P < 0.001), HER2 overexpression (P < 0.001) and negative hormone receptor status (P < 0.001). ALDH1 expression was observed in 14% of DCISs but was not correlated with any clinicopathological parameter. The IDC patients were followed up for 7–190 months (median: 120 months), and groups with ALDH1 expression had shorter relapse‐free survival (P = 0.0013) and overall survival (OS) (P = 0.0005) by log‐rank test. By Cox’s multivariate analysis, it had a weak effect on OS (P = 0.047), and its most significant effect on OS was observed in node‐positive groups (P = 0.013). No significant differences in OS stratified by ALDH1 expression status in lymph node metastases were noted. Conclusions: ALDH1 expression in primary cancer is an independent prognostic factor in node‐positive breast cancer patients.     

Tumour budding and other prognostic pathological features at invasive margins in serrated colorectal adenocarcinoma: a comparative study with conventional carcinoma

García‐Solano J, Conesa‐Zamora P, Trujillo‐Santos J, Mäkinen M J & Pérez‐Guillermo M
(2011) Histopathology  59 , 1046–1056
Tumour budding and other prognostic pathological features at invasive margins in serrated colorectal adenocarcinoma: a comparative study with conventional carcinoma Aims: To assess the incidence of tumour budding (TB), cytoplasmic pseudo‐fragments (CyPs), tumour growth pattern (TGP) and peritumoural lymphocytic infiltration (PLI) in a series of serrated adenocarcinoma (SAC) and conventional carcinomas (CCs) of the colorectum in order to ascertain whether such features could explain the worse prognosis of SAC and whether they have prognostic value in SACs. Methods and results: Tumour budding, CyPs, TGP and PLI were evaluated in 81 SACs and 81 matched CCs. Kaplan–Meier survival curves and Cox logistic regression analysis were obtained for histological parameters. SACs had more high‐grade (HG) TB (HG‐TB) (69.1%), HG‐CyPs (47%), infiltrative TGP (42%) and weak PLI (W‐PLI) (65.4%) than CCs (40.7%, P = 0.0003; 19.7%, P = 0.0002; 29.7%, P = 0.07; 45.7%, P = 0.0087). SACs with HG‐TB (P = 0.017), HG‐CyPs (P = 0.045), infiltrating TGP (P < 0.001) and W‐PLI (P = 0.04) had a worse 5‐year survival, as had SACs with infiltrating TGP (P = 0.047) and W‐PLI (P = 0.04) compared with CCs. For SACs, infiltrative TGP and W‐PLI were independent prognostic parameters on multivariate analysis, as was location and regional node status. Conclusion: Compared to CC, SAC displayed more HG‐TB, HG‐CyPs and fewer PLI at the invasive margins and this may account for its poorer clinical outcome. TB, CyPs, TGP and PLI are useful histological prognostic aids in SAC.     

Spindle proteins in resected pancreatic head adenocarcinomas: BubR1 is an independent prognostic factor in pancreatobiliary‐type tumours

Gladhaug I P, Westgaard A, Schjølberg A R, Burum‐Auensen E, Pomianowska E & Clausen O P F
(2010) Histopathology 56, 345–355 Spindle proteins in resected pancreatic head adenocarcinomas: BubR1 is an independent prognostic factor in pancreatobiliary‐type tumours Aims: Spindle proteins such as Aurora A, Mad2 and BubR1 are important for chromosome segregation during mitosis. Dysfunction of these proteins is implicated in the development of many cancers. The aim was to examine their possible prognostic impact in resected adenocarcinomas in the pancreatic head. Methods and results: Two hundred and eighteen consecutively resected pancreatobiliary‐type (n = 145) and intestinal‐type (n = 73) adenocarcinomas involving the pancreatic head were examined for expression of Aurora A, Mad2 and BubR1 by immunohistochemistry on tissue microarrays. Aurora A (P < 0.001) and Mad2 (P = 0.003) were expressed more often and at higher levels in intestinal‐type compared with pancreatobiliary‐type tumours, whereas BubR1 was equally expressed in both histological types. Expression of BubR1, Aurora A and Mad2 was not associated with ploidy status. None of the spindle proteins was significantly associated with prognosis in intestinal‐type tumours. In pancreatobiliary‐type tumours, any BubR1 expression was sufficient to predict poor prognosis (P = 0.006), whereas Aurora A and Mad2 expression was not significantly associated with prognosis (P = 0.86 and P = 0.87, respectively). On adjusted Cox regression analysis, BubR1 expression independently predicted poor prognosis [P =0.002; hazard ratio (HR) 1.87, 95% confidence interval (CI) 1.26, 2.79)], particularly in small tumours (P = 0.001; HR 2.93, 95% CI 1.53, 5.62). Conclusion: BubR1 expression is a novel, independent adverse prognostic factor after pancreatoduodenectomy of pancreatobiliary‐type adenocarcinomas.     

Cytoplasmic expression of survivin is an independent predictor of poor prognosis in patients with salivary gland cancer

Stenner M, Weinell A, Ponert T, Hardt A, Hahn M, Preuss S F, Guntinas‐Lichius O & Klussmann J P
(2010) Histopathology 57 , 699–706
Cytoplasmic expression of survivin is an independent predictor of poor prognosis in patients with salivary gland cancer Aims: The expression of the inhibitor of apoptosis protein survivin has been shown to be a significant prognostic indicator in various human cancers. The aim was to assess its expression and prognostic value in salivary gland adenocarcinoma and muco‐epidermoid carcinoma. Methods and results: Survivin expression was analysed in 48 patients with parotid gland cancer (21 muco‐epidermoid, 27 adenocarcinomas) by means of immunohistochemistry. The experimental findings were correlated with clinicopathological and survival parameters. A high cytoplasmic expression of survivin was found in 30% of the examined tumours without any significant correlation with the patients’ clinicopathological characteristics (P > 0.05). Within all patients, the estimated overall survival rate of muco‐epidermoid carcinomas was significantly better than that of adenocarcinomas (P = 0.013). A high cytoplasmic survivin expression significantly indicated a poor 5‐year disease‐free survival rate compared to patients with a low cytoplasmic survivin expression in the whole group (P = 0.001) and in adenocarcinomas (P = 0.004). In a multivariate analysis, a high cytoplasmic survivin expression was the only independent prognostic indicator for a significantly poorer 5‐year disease‐free survival rate (P = 0.001). Conclusions: The correlation between cytoplasmic survivin expression and survival in salivary gland malignancies might make this an effective tool in patient follow‐up, prognosis and targeted therapy in future.