Publications

2010

STAMP1 Is Both a Proliferative and an Antiapoptotic Factor in Prostate Cancer.

Wang L, Jin Y, Arnoldussen YJ, Jonson I, Qu S, Mælandsmo GM, Kristian A, Risberg B, Wæhre H, Danielsen HE, Saatcioglu F.
Cancer Res. 2010 Jun 29. [Epub ahead of print]

PMID: 20587517

2009

Identification of p53 as a strong predictor of survival for patients with malignant peripheral nerve sheath tumors.

Brekke HR, Kolberg M, Skotheim RI, Hall KS, Bjerkehagen B, Risberg B, Domanski HA, Mandahl N, Liestøl K, Smeland S, Danielsen HE, Mertens F, Lothe RA.
Neuro Oncol. 2009 Oct;11(5):514-28. Epub 2009 Jan 30.

Abstract

The purpose of this study was to identify new prognostic biomarkers with clinical impact in malignant peripheral nerve sheath tumor (MPNST), a highly aggressive malignancy for which no consensus therapy exists besides surgery. We have used tissue microarrays (TMAs) to assess in situ expression of 14 cell-cycle-regulating proteins in 64 well-characterized MPNST patients: 36 sporadic and 28 with neurofibromatosis type 1 (NF1). We developed a new software application for evaluation and logistics of the TMA images and performed a literature survey of cell cycle proteins in MPNST. For NF1-associated patients, there was a clear association between nuclear expression of p53 and poor survival (p = 0.004). Among the other proteins analyzed, we also found significant associations between survival and clinical variables, but none were as strong as that for p53. For the total series of MPNSTs, p53 was shown to be an independent predictor of survival, and patients without remission, with tumor size larger than 8 cm, and with positive p53 expression had a 60 times greater risk of dying within the first 5 years compared with the remaining patients (p = 0.000002). This is the most comprehensive study of in situ protein expression in MPNST so far, and expressed p53 was found to be a strong surrogate marker for outcome. Patients in complete remission with a primary p53-positive MPNST diagnosis may be considered in a high-risk subgroup and candidates for adjuvant treatment.

PMID: 19182148

Large scale genomic instability as an additive prognostic marker in early prostate cancer.

Pretorius ME, Waehre H, Abeler VM, Davidson B, Vlatkovic L, Lothe RA, Giercksky KE, Danielsen HE.
Cell Oncol. 2009;31(4):251-9.

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.

PMID: 19633362

Large-scale genomic instability in colon adenocarcinomas and correlation with patient outcome.

Bondi J, Pretorius M, Bukholm I, Danielsen H.
APMIS. 2009 Oct;117(10):730-6.

Abstract

The purpose of this study was to evaluate the association between DNA content in colon adenocarcinomas using high-resolution image cytometry and patient outcome. Tumours from 219 patients operated for colon adenocarcinoma were analysed using high-resolution image cytometry. Proteins involved in cell cycle propulsion (cyclins A, D1, D3 and E) and cell proliferation (c-Myc and non-membranous beta-catenin) have previously been reported in the same cohort and were included in this study. The results were related to disease-free survival and to cancer-specific death. Patients with aneuploid tumours showed shorter relapse-free survival than patients with euploid tumours (univariate log-rank test, p = 0.004 and multivariate Cox regression model p = 0.009, HR 0.51, 95% CI 0.31-0.84). Also the risk of death from cancer was greater in patients with aneuploid tumours (log-rank test, p = 0.006 multivariate Cox regression model p = 0.014, HR 0.47, 95% CI 0.26-0.86). When analysing patients with Dukes stages A and B, nuclear expression of beta-catenin was highly significantly associated with both shorter relapse-free survival (p < 0.005, HR 5.0, 95% CI 1.6-15.5) and cancer-specific death (p = 0.036, HR 6.9, 95% CI 1.1-42.1). DNA content in colon adenocarcinomas measured by image cytometry is an independent predictor of prognosis in our patients operated for colon adenocarcinoma.

PMID: 19775341

Beta-catenin expression in uterine sarcomas and its relation to clinicopathological parameters.

Kildal W, Pradhan M, Abeler VM, Kristensen GB, Danielsen HE.
Eur J Cancer. 2009 Sep;45(13):2412-7. Epub 2009 Jul 20.

Abstract

Aberrations in the Wnt/beta-catenin signalling pathway are suggested as mediators of chromosomal instability and carcinogenesis. beta-catenin acts both as a component of the membranous adhesion system, and as a transcription activator in the nucleus. beta-Catenin immunoreactivity was evaluated in 353 uterine sarcomas (US) including 231 leiomyosarcomas (LMS), 82 endometrial stromal sarcomas (ESS), 22 adenosarcomas (AS) and 18 undifferentiated uterine sarcomas (UUS). Up-regulated membranous beta-catenin was observed in 25% of the LMS (p=0.039), 21% of the ESS (p=0.072) and 39% of the UUS (p=0.025). Cytoplasmic beta-catenin was up-regulated in 36% of the LMS (p=0.008) and 33% of the UUS (p=0.028). Nuclear beta-catenin expression was observed in 23% of the LMS (p=0.051), 61% of ESS (p=0.628) and in the sarcoma component of 68% of the AS. In patients with LMS, membranous beta-catenin was associated with poor crude survival in univariate (p=0.045), but not in multivariate analyses. In patients with ESS, nuclear beta-catenin expression was related to spread of tumour (p=0.033), but not to survival. The observation of up-regulated beta-catenin expression in US might suggest a so far undocumented role for the Wnt/beta-catenin pathway in these malignancies.

PMID: 19622417

Gross genomic alterations differ between serous borderline tumors and serous adenocarcinomas–an image cytometric DNA ploidy analysis of 307 cases with histogenetic implications.

Pradhan M, Davidson B, Tropé CG, Danielsen HE, Abeler VM, Risberg B.
Virchows Arch. 2009 Jun;454(6):677-83. Epub 2009 May 8.

Abstract

Our objective was to study the gross genomic alterations in serous borderline tumors and serous adenocarcinomas of the ovary. A retrospective analysis of 245 serous borderline tumors and 62 serous adenocarcinomas from 249 patients was performed using high-resolution image cytometric DNA ploidy analysis. DNA ploidy status, S-phase fraction, and DNA index were evaluated. The majority of serous borderline tumors were diploid (225/245 cases, 92%). The remaining 8% showed an aneuploid peak predominantly with DNA index of less than 1.4. Grades 2 and 3 serous adenocarcinomas were more often (80%) nondiploid, mostly with DNA index exceeding 1.4. Grade 1 serous adenocarcinomas were an intermediate group, more similar to serous borderline tumors. The S-phase fraction increased from serous borderline tumors (mean = 0.6%) through grade 1 serous adenocarcinomas (mean = 2.8%), being highest in grades 2 and 3 adenocarcinomas (mean = 6.8%). Our findings support the hypothesis that serous borderline tumors and grades 2 and 3 serous adenocarcinomas are genomically different lesions, with grade 1 serous adenocarcinomas being an intermediate group more close to borderline tumors.

PMID: 19421773

The prognostic value of DNA ploidy in a total population of uterine sarcomas.

Kildal W, Abeler VM, Kristensen GB, Jenstad M, Thoresen SØ, Danielsen HE.
Ann Oncol. 2009 Jun;20(6):1037-41. Epub 2009 Feb 6.

BACKGROUND

The diagnosis of uterine sarcoma is associated with poor outcome for the patient and there is a need for reliable prognostic markers. Most previous studies on the prognostic value of DNA ploidy include few uterine sarcomas and report conflicting results.

Materials and methods

We examined the prognostic value of DNA ploidy and its association with clinicopathological parameters and crude survival in a total population of 354 sarcoma.

RESULTS

In univariate analyses, we observed significantly better crude survival for endometrial stromal sarcomas (ESS) and adenosarcoma (AS) patients with diploid as compared with nondiploid tumors, but not for patients with leiomyosarcomas (LMS). In Cox multivariate analyses, DNA ploidy was the only significant predictor of survival for patients with AS. In LMS, mitotic index (MI), tumor size, tumor extent and tumor margins, whereas for ESS, MI, tumor extent and tumor necrosis obtained independent significance of survival. DNA ploidy was a significant predictor of survival for LMS patients in Cox regression analyses when excluding MI.

CONCLUSION

DNA ploidy might be useful as a prognostic marker in patients with LMS and AS.

PMID: 19201782

Uterine sarcomas in Norway. A histopathological and prognostic survey of a total population from 1970 to 2000 including 419 patients.

Abeler VM, Røyne O, Thoresen S, Danielsen HE, Nesland JM, Kristensen GB.
Histopathology. 2009 Feb;54(3):355-64.

AIMS

To determine the frequency and survival of the various types of uterine sarcoma in the total population of Norway and evaluate histopathological prognostic factors in order to identify risk groups.

METHODS AND RESULTS

Histopathological review of all uterine sarcoma cases reported to the Norwegian Cancer Registry during 1970-2000 was undertaken. Survival dates were provided by The Cancer Registry. Kaplan-Meier survival curves were generated. The log rank test was used for univariate analysis and a Cox proportional hazards regression model for multivariate evaluation of survival. Stage of disease was the most important prognostic factor for all tumour types. Tumour size and the mitotic index (MI) were significant prognostic factors (P < 0.0001) in leiomyosarcomas confined to the uterus and allowed for separation into three risk groups with marked differences in prognosis. The prognosis of endometrial stromal sarcomas confined to the uterus was related to MI (P < 0.0001) and tumour cell necrosis (P < 0.004). Combining these parameters allowed for separation into three risk groups with marked difference in prognosis. In adenosarcomas, tumour cell necrosis was the only significant prognostic factor.

CONCLUSIONS

There are marked differences in survival between uterine sarcoma types. Leiomyosarcomas and endometrial stromal sarcomas can be divided into different groups.

PMID: 19236512

2008

The mitogen-activated protein kinase phosphatase vaccinia H1-related protein inhibits apoptosis in prostate cancer cells and is overexpressed in prostate cancer.

Arnoldussen YJ, Lorenzo PI, Pretorius ME, Waehre H, Risberg B, Maelandsmo GM, Danielsen HE, Saatcioglu F.
Cancer Res. 2008 Nov 15;68(22):9255-64.

Abstract

Androgen ablation during the initial stages of prostate cancer causes regression of the tumor due to an increase in apoptosis and reduced cellular proliferation. However, prostate cancer invariably progresses to an androgen-independent state for poorly understood reasons. Previous studies showed that c-Jun NH(2) terminal kinase (JNK) is required for 12-O-tetradecanoylphorbol-13-acetate (TPA)- and thapsigargin (TG)-induced apoptosis in the androgen-responsive prostate cancer cell line LNCaP. Androgens protect LNCaP cells from TPA-induced or TG-induced apoptosis via down-regulation of JNK activation. However, the molecular mechanisms of this inhibition are not clear. Here, we systematically investigated the possible regulation of mitogen-activated protein kinase phosphatases/dual-specificity phosphatases during apoptosis of LNCaP cells and found that Vaccinia H1-related protein (VHR/DUSP3) is up-regulated by androgens during inhibition of apoptosis in LNCaP cells, but not in androgen-independent DU145 cells. Ectopic expression of wild-type VHR, but not a catalytically inactive mutant, interfered with TPA- and TG-induced apoptosis. Consistently, small interfering RNA-mediated knockdown of endogenous VHR increased apoptosis in response to TPA or TG in the presence of androgens. Furthermore, COS7 cells stably expressing wild-type VHR, but not a mutant, had a decrease in JNK phosphorylation. In vivo, VHR expression decreased in the androgen-dependent human prostate cancer xenograft CWR22 upon androgen withdrawal and was inversely correlated to JNK phosphorylation. Expression analysis in human prostate cancer specimens showed that VHR is increased in prostate cancer compared with normal prostate. These data show that VHR has a direct role in the inhibition of JNK-dependent apoptosis in LNCaP cells and may therefore have a role in prostate cancer progression.

PMID: 19010898

Statistical nuclear texture analysis in cancer research: a review of methods and applications.

Nielsen B, Albregtsen F, Danielsen HE.
Crit Rev Oncog. 2008;14(2-3):89-164.

Abstract

In digital pathology, the field of nuclear texture analysis gives information about the spatial arrangement of the pixel gray levels in a digitized microscopic nuclear image, providing statistical texture measures that may be used as quantitative tools for diagnosis and prognosis of human cancer. In the present work, we have reviewed nuclear texture analysis in human cancer research, with emphasis on (i) statistical texture analysis methods, (ii) methods for feature evaluation and feature set selection, (iii) classification methods and error estimation, and (iv) the recent literature in the field, focusing on diagnosis- and prognosis-related applications. The application study covers the period from 1995 to 2007. In order to find nuclear features that discriminate robustly between cases from different diagnostic or prognostic classes, a statistical evaluation of features must be performed, and this demands careful experimental design. The present review reveals that it is quite common to evaluate a large number of features on a limited learning set of clinical material, without testing the chosen classifier on an independent validation data set. This easily leads to overoptimistic results. Out of 160 papers, we found only 30 papers in which the classifier was evaluated on an independent validation data set. Even in these studies, some good results have been hampered by small validation groups. However, it is encouraging to note that those publications meeting the requirements of an optimal study are generally showing good results. Thus, it is well documented that nuclear texture analysis is showing promising results as a novel diagnostic and/or prognostic marker. Hopefully, we will soon see that these promising studies will be replicated in large, prospective, multicenter trials.

PMID: 19409060

Consistent numerical chromosome aberrations in thecofibromas of the ovary.

Micci F, Haugom L, Abeler VM, Tropé CG, Danielsen HE, Heim S.
Virchows Arch. 2008 Mar;452(3):269-76.

Abstract

Sex cord-stromal tumors of the ovary comprise 8% of all ovarian neoplasms. Because they consist of cells that resemble embryonic sex cord and/or specialized ovarian stroma cells, their cytologic and histologic features can be viewed as reflecting a continuum from fibromas to thecomas with thecofibromas in between. Existing cytogenetic knowledge about ovarian thecomas-thecofibromas-fibromas is restricted to 44 cases with chromosomal abnormalities. The most common aberration has been trisomy 12, identified either by karyotyping or using fluorescence in situ hybridization (FISH). We wanted to obtain more information about the genomic composition of these tumors, and, therefore, examined 29 new thecoma-thecofibroma-fibroma tumors of the ovary using karyotyping, comparative genomic hybridization, interphase FISH, and DNA ploidy analysis. We detected aneuploidy in 21 tumors. Trisomy and/or tetrasomy 12 was the most common chromosomal aberration, found in 15 tumors (71.5% of the aneuploid tumors or 51.5% of all analyzed tumors), followed by trisomy for chromosomes 10, 18, 4, and 9. Some monosomies (for chromosomes 4, 9, 10, and 18) were also identified, either as the sole change or in combination with trisomies. The nonrandom occurrence of these aneuploidies in these benign tumors strongly indicates that they play a major pathogenetic role, but how trisomies and other aneuploidies contribute to tumorigenesis remains unknown.

PMID: 18188592

2007

A distinct pattern in the DNA ploidy histograms of hydatidiform moles and nonmolar abortuses is caused by accumulation of trophoblasts in the late s-phase.

Pradhan M, Abeler VM, Danielsen HE, Risberg B.
Int J Gynecol Pathol. 2007 Oct;26(4):432-6.

Abstract

DNA ploidy analysis is a useful tool to distinguish the partial hydatidiform moles (PMs) from complete hydatidiform moles (CMs) and nonmolar abortuses (NAs). DNA ploidy histograms of hydatidiform moles are sometimes difficult to interpret because of the uneven distribution of nuclei in the S-phase, simulating aneuploid peaks. In this study, we analyzed DNA ploidy histograms of 25 CMs, 16 PMs, and 28 NAs, with special reference to the accumulation of cells in the late S-phase using a high-resolution DNA image cytometry. All the gestational products demonstrated the accumulation of cells in the late part of the S-phase fraction. To objectify the observation, we compared the percentage of cells in the second quarter with that of the third quarter of the S-phase fraction. All the gestational products had significantly lower (P < 0.001) percentage of cells in the second compared with that of the third quarter of the S-phase. The mean ratios of the third quarter to the second quarter in CMs, PMs, and NAs were 1.9, 1.7, and 2.5, respectively. This was significantly different from that of highly proliferative endometrial carcinomas. The knowledge of this specific S-phase fraction distribution in molar and nonmolar pregnancy material is important when interpreting the DNA histograms. The possibility of hypoxia being the cause of this phenomenon is also discussed.

PMID: 17885494

A distinct pattern in the DNA ploidy histograms of hydatidiform moles and nonmolar abortuses is caused by accumulation of trophoblasts in the late s-phase.

Pradhan M, Abeler VM, Danielsen HE, Risberg B.
Int J Gynecol Pathol. 2007 Oct;26(4):432-6.

Abstract

DNA ploidy analysis is a useful tool to distinguish the partial hydatidiform moles (PMs) from complete hydatidiform moles (CMs) and nonmolar abortuses (NAs). DNA ploidy histograms of hydatidiform moles are sometimes difficult to interpret because of the uneven distribution of nuclei in the S-phase, simulating aneuploid peaks. In this study, we analyzed DNA ploidy histograms of 25 CMs, 16 PMs, and 28 NAs, with special reference to the accumulation of cells in the late S-phase using a high-resolution DNA image cytometry. All the gestational products demonstrated the accumulation of cells in the late part of the S-phase fraction. To objectify the observation, we compared the percentage of cells in the second quarter with that of the third quarter of the S-phase fraction. All the gestational products had significantly lower (P < 0.001) percentage of cells in the second compared with that of the third quarter of the S-phase. The mean ratios of the third quarter to the second quarter in CMs, PMs, and NAs were 1.9, 1.7, and 2.5, respectively. This was significantly different from that of highly proliferative endometrial carcinomas. The knowledge of this specific S-phase fraction distribution in molar and nonmolar pregnancy material is important when interpreting the DNA histograms. The possibility of hypoxia being the cause of this phenomenon is also discussed.

PMID: 17885494

Kallikrein 4 is a proliferative factor that is overexpressed in prostate cancer.

Klokk TI, Kilander A, Xi Z, Waehre H, Risberg B, Danielsen HE, Saatcioglu F.

Cancer Res. 2007 Jun 1;67(11):5221-30.

Abstract

Kallikrein 4 (KLK4) is a member of the human tissue KLK family. Whereas all other KLKs are secreted proteins with extracellular functions, KLK4 is primarily localized to the nucleus, indicating that it has a different function compared with other members of the KLK family. In addition, KLK4 expression is highly enriched in the prostate and is regulated by androgens. Here, we studied the possible functional role of KLK4 in prostate cancer cells and examined its expression at the protein level in prostate cancer specimens. Consistent with its mRNA expression, KLK4 protein is significantly overexpressed in malignant prostate compared with normal prostate. KLK4 expression is predominantly in the nucleus of basal cells in the prostate epithelium in keeping with its distribution in prostate cancer cells in vitro. Furthermore, adenovirus-mediated expression of KLK4 dramatically induces proliferation of prostate cancer cells, at least in part through significant alterations in cell cycle regulatory gene expression. Consistent with these data, small interfering RNA-mediated knockdown of endogenous KLK4 in LNCaP prostate cancer cells inhibits cell growth. These data identify KLK4 as the first member of the KLK family that is a proliferative factor with effects on gene expression and indicate that it may have an important role in prostate cancer development and progression.

PMID: 17545602

Kallikrein 4 is a proliferative factor that is overexpressed in prostate cancer.

Klokk TI, Kilander A, Xi Z, Waehre H, Risberg B, Danielsen HE, Saatcioglu F.
Cancer Res. 2007 Jun 1;67(11):5221-30.

Abstract

Kallikrein 4 (KLK4) is a member of the human tissue KLK family. Whereas all other KLKs are secreted proteins with extracellular functions, KLK4 is primarily localized to the nucleus, indicating that it has a different function compared with other members of the KLK family. In addition, KLK4 expression is highly enriched in the prostate and is regulated by androgens. Here, we studied the possible functional role of KLK4 in prostate cancer cells and examined its expression at the protein level in prostate cancer specimens. Consistent with its mRNA expression, KLK4 protein is significantly overexpressed in malignant prostate compared with normal prostate. KLK4 expression is predominantly in the nucleus of basal cells in the prostate epithelium in keeping with its distribution in prostate cancer cells in vitro. Furthermore, adenovirus-mediated expression of KLK4 dramatically induces proliferation of prostate cancer cells, at least in part through significant alterations in cell cycle regulatory gene expression. Consistent with these data, small interfering RNA-mediated knockdown of endogenous KLK4 in LNCaP prostate cancer cells inhibits cell growth. These data identify KLK4 as the first member of the KLK family that is a proliferative factor with effects on gene expression and indicate that it may have an important role in prostate cancer development and progression.

PMID: 17545602