Analytical Cellular Pathology - Volume 31, issue 4

Authors: Pretorius, Maria E. | Wæhre, Håkon | Abeler, Vera M. | Davidson, Ben | Vlatkovic, Ljiljana | Lothe, Ragnhild A. | Giercksky, Karl-Erik | Danielsen, Håvard E.

Article Type: Research Article

Abstract: Background: The clinical outcome for the individual prostate cancer patient is often difficult to predict, due to lack of reliable independent prognostic biomarkers. We tested DNA ploidy as a prognostic factor for clinical outcome in 186 patients treated with radical prostatectomy. Methods: DNA ploidy was measured using an automatic image cytometry system and correlated with preoperative PSA, age at surgery, Mostofi grade, surgical margins and Gleason score. Results: The mean follow up time after operation was 73.3 months (range 2–176 months). Of the 186 prostatectomies, 96 were identified as diploid, 61 as tetraploid and 29 as aneuploid. Twenty-three …per cent, 36% and 62% of the diploid, tetraploid and aneuploid cases respectively, suffered from relapse during the observation time. DNA ploidy, Gleason score, Mostofi grading, surgical margins and preoperative PSA were all significant predictors of relapse in a univariate analysis. On multivariate analysis, only Gleason score and DNA ploidy proved to be independently predictors of disease recurrence. Furthermore, among the 68 cases identified with Gleason score 7, DNA ploidy was the only significant predictor of disease recurrence. Conclusions: Our data suggest that DNA ploidy should be included as an important additive prognostic factor for prostate cancer, especially for patients identified with Gleason score 7 tumours. Show more

Keywords: DNA ploidy, Gleason, prognosis, prostate cancer, radical prostatectomy

DOI: 10.3233/CLO-2009-0463

Citation: Analytical Cellular Pathology, vol. 31, no. 4, pp. 251-259, 2009

Authors: Skaland, Ivar | Janssen, Emiel A.M. | Gudlaugsson, Einar | Hui Ru Guo, Lydia | Baak, Jan P.A.

Article Type: Research Article

Abstract: Purpose: Prognostic comparison of phosphohistone-H3 (PPH3) with Cytokeratin 5/6 and/or 14 positive (=basal-CK), triple (ER, PR, HER2)-negative (=TNP) and basal-like (=TNP and basal-CK positive) phenotype in invasive breast cancers. Patients and methods: Classical variables, PPH3, ER, PR, basal-CK and HER2 in 240 T1–2 N0 M0 patients under 71 years. Results: TNP and basal-like cancers had higher PPH3 expression than the other cancers (mean 48 versus 11, P<0.001). Fifteen percent of the patients in the whole group, but 32–38% of TNP and basal-like cancers recurred. With multivariate analysis, PPH3<13 (n=156) versus ≥13 (n=84=35% of all cases) was the …strongest and only prognosticator (10-year survival 96% and 64%, P≤0.001, Hazard ratio=9.0). Conclusion: PPH3 is the strongest prognosticators in luminal, Triple negative and basal-like T1–2 N0 M0 invasive breast cancers. Show more

Keywords: Breast cancer, prognosis, proliferation, Phosphohistone H3, basal-like

DOI: 10.3233/CLO-2009-0464

Citation: Analytical Cellular Pathology, vol. 31, no. 4, pp. 261-271, 2009

Authors: Wang, Shouye | Basson, Marc D.

Article Type: Research Article

Abstract: Cell adhesion is important in cancer metastasis. Malignant cells in cancer patients may be exposed to physical forces such as extracellular pressure and shear, that stimulate their adhesion to matrix proteins, endothelium and surgical wounds. Pressure induces phosphorylation of AKT and focal adhesion kinase (FAK), which are required for pressure-stimulated cancer cell adhesion, but what mediates this effect is unknown. ILK may influence cell adhesion and FAK and AKT phosphorylation in other settings. We therefore hypothesized that ILK might also regulate pressure-stimulated cancer cell adhesion through AKT and FAK phosphorylation. Silencing ILK by siRNA reduced basal cancer cell adhesion and …prevented the stimulation of adhesion by pressure. ILK mediated pressure-stimulated adhesion through specifically regulating phosphorylation of AKT at Ser473 and FAK at Tyr397 and 576 as well as ILK association with FAK and AKT. The siRNA-mediated loss of function of ILK in regulating increase in adhesion by pressure was not rescued by overexpression of α-parvin, an important ILK binding partner, although pressure promoted ILK–α-parvin association and translocated both ILK and α-parvin from cytosol to membrane/cytoskeleton. ILK may be a key mediator of mechanotransduced signals in cancer cells and an important therapeutic target to inhibit metastatic cancer cell adhesion. Show more

Keywords: Cell adhesion, integrin-linked kinase (ILK), alpha-parvin/actopaxin/CH-ILKBP, AKT/protein kinase B, focal adhesion kinase (FAK), extracellular pressure, phosphorylation

DOI: 10.3233/CLO-2009-0469

Citation: Analytical Cellular Pathology, vol. 31, no. 4, pp. 273-289, 2009

Authors: Smeets, Serge J. | Brakenhoff, Ruud H. | Ylstra, Bauke | van Wieringen, Wessel N. | van de Wiel, Mark A. | Leemans, C. René | Braakhuis, Boudewijn J.M.

Article Type: Research Article

Abstract: The common risk factors for oral and oropharyngeal cancer are tobacco smoking and alcohol consumption, and recently the human papillomavirus (HPV) was shown to be involved in a subgroup. HPV-positive and -negative carcinomas can be distinguished on basis of their genetic profiles. Aim of this study was to investigate patterns of chromosomal aberrations of HPV-negative oral and oropharyngeal squamous cell carcinomas (OOSCC) in order to improve stratification of patients regarding outcome. Thirty-nine OOSCCs were classified on basis of their genetic pattern determined by array comparative genomic hybridization (aCGH). Resulting groups were related to patient and tumor characteristics using the Fisher's …exact test and in addition to survival with the Kaplan–Meier and log rank tests. Classification distinguished three groups, one characterized by hardly any chromosomal aberration (N=8) and another by a relatively high level (N=26), and one with a very high level (N=5) of chromosomal aberrations. This classification was significantly (p=0.003) associated with survival, with the best survival in the genetically ‘silent’ group and the worst survival in the most aberrant group. The silent profile was significantly (p<0.05) associated with wild-type TP53, an absence of alcohol consumption and a female gender. These carcinomas were negative for microsatellite instability. This classification of OOSCC was confirmed in an independent set of 89 oral carcinomas. In conclusion, the discovery of these new classes of oral and oropharyngeal cancer with unique genetic and clinical characteristics has important consequences for future basic and clinical studies. Show more

Keywords: Array CGH, head and neck cancer, HPV, oral cancer, prognosis, TP53

DOI: 10.3233/CLO-2009-0471

Citation: Analytical Cellular Pathology, vol. 31, no. 4, pp. 291-300, 2009

Authors: Schrödl, Kathrin | Oelmez, Hamza | Edelmann, Martin | Huber, Rudolf Maria | Bergner, Albrecht

Article Type: Research Article

Abstract: Background: Chemotherapy often leads to encouraging responses in lung cancer. But, in the course of the treatment, resistance to chemotherapy ultimately limits the life expectancy of the patient. We aimed at investigating if treatment with cisplatin alters the intracellular Ca2+ -homeostasis of lung cancer cells and how this may be related to cisplatin resistance. Methods: The squamous cell lung carcinoma cell line EPLC M1 and the small cell lung cancer cell line H1339 were exposed to cisplatin analogue to the in vivo pharmacokinetics. Changes in the cytoplasmic Ca2+ -concentration ([Ca2+ ]c ) were recorded using fluorescence microscopy. Protein expression …was quantified using immuno-fluorescence and Western Blot analysis. Changes in gene expression were accomplished by small-interfering (si) RNA techniques. Results: Four “cycles” of cisplatin led to low level resistance in EPLC and H1339 cells. In the low level resistant cell clones, the Ca2+ -content of the endoplasmic reticulum (ER) was decreased. In low level resistant EPLC cells, this was correlated with an increased expression of the inositol-1,4,5-trisphosphate receptor (IP3 R). Inhibiting the increased expression of IP3 R using siRNA, the low level resistance could be reversed. In low level resistant H1339 cells, the decreased Ca2+ -content of the ER was correlated with a decreased expression of sarco/endoplasmic reticulum Ca2+ -ATPases (SERCA). Decreasing the expression of SERCA in naïve H1339 cells resulted in low level cisplatin resistance. Conclusion: We conclude that cisplatin therapy leads to a decreased Ca2+ -content of the ER thereby inducing low level resistance. This is caused by upregulation of the IP3 R in EPLC and decreased expression of SERCA in H1339 cells. Show more

Keywords: Lung cancer, cisplatin, calcium, SERCA, IP_3R

DOI: 10.3233/CLO-2009-0472

Citation: Analytical Cellular Pathology, vol. 31, no. 4, pp. 301-315, 2009

Authors: Graveland, A. Peggy | de Maaker, Michiel | Braakhuis, Boudewijn J.M. | de Bree, Remco | Eerenstein, Simone E.J. | Bloemena, Elisabeth | Leemans, C. René | Brakenhoff, Ruud H.

Article Type: Research Article

Abstract: A great disappointment in head and neck cancer surgery is that 10–30% of head and neck squamous cell carcinoma (HNSCC) patients develop local recurrences despite histopathologically tumor-free surgical margins. These recurrences result from either minimal residual cancer (MRC) or preneoplastic lesions that remain behind after tumor resection. Distinguishing MRC from preneoplasic lesions is important to tailor postoperative radiotherapy more adequately. Here we investigated the suitability of quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) using human Ly-6D (hLy-6D) transcripts as molecular marker to detect MRC in surgical margins. Submucosal samples of deep surgical margins were collected from 18 non-cancer control patients …and 67 HNSCC patients of whom eight had tumor-positive surgical margins. The samples were analyzed with hLy-6D qRT-PCR, and the data were analyzed in relation to the clinicohistological parameters. A significant difference was shown between the group of patients with histopathological tumor-positive surgical margins and the non-cancer control group (p<0.001), and the group of patients with histopathological tumor-free surgical margins (p=0.001). This study shows a novel approach for molecular analysis of deep surgical margins in head and neck cancer surgery. The preliminary data of this approach for detection of MRC in deep margins of HNSCC patients are promising. Show more

Keywords: Head and neck cancer, hLy-6D, minimal residual cancer, molecular diagnosis, qRT-PCR, surgical margin

DOI: 10.3233/CLO-2009-0474

Citation: Analytical Cellular Pathology, vol. 31, no. 4, pp. 317-328, 2009