Supplementary Figures S1-S2 from ABCC2 (MRP2, cMOAT) Can Be Localized in the Nuclear Membrane of Ovarian Carcinomas and Correlates with Resistance to Cisplatin and Clinical Outcome

Telemedizin, definiert als ärztliche Leistungserbringung über räumliche oder zeitliche Distanz hinweg und unter Einsatz moderner Informations- und Kommunikationstechnologien, gewinnt auch in der Versorgung von Krebspatienten an Bedeutung. Die onkologische Versorgung in Deutschland wird maßgeblich von den zertifizierten Zentren der Deutschen Krebsgesellschaft geprägt. Die Zentren erfüllen hohe Qualitätsanforderungen, beispielsweise mit Blick auf die fachliche Expertise und interdisziplinäre Zusammenarbeit. Der so bestehende große Kommunikationsbedarf eröffnet potenzielle Einsatzfelder für die Telemedizin. Anhand der Beispiele Tele-Tumorkonferenz und Telepathologie beschreibt dieser Beitrag Chancen und Risiken bei der Implementierung von Telemedizin in der Onkologie: Sie ermöglicht den Leistungserbringern eine effizientere Zusammenarbeit und die flächendeckende Einbringung onkologischen Wissens. Andererseits besteht je nach Einsatzbereich und Umsetzung die Gefahr von Informationsverlusten oder Einbußen für den Versorgungsstandard. Diese Aspekte gilt es kritisch abzuwägen, damit die Telemedizin die Onkologie positiv weiterentwickelt und gleichzeitig die Qualität der Patientenversorgung erhalten bleibt.

e23141 Background: Cancer profiling is receiving increased attention, but comprehensive algorithms to guide targeted and immunotherapies for patients are scarce and complex biomarker data are not easily accessible to clinicians. Methods: Databases (TARGET database, mycancergenome.org, pct.mdanderson.org) and pubmed (search terms “predictive biomarker” AND “cancer”) were queried to identify a gene list. For assessment of pharmacogenomic implications, public databases (CIViC at civic.genome.wustl.edu/, mycancergenome.org) and pubmed.com (search terms “GENE NAME” AND “treatment” AND “cancer”) were searched for each gene. Evidence was added to the CIViC database as applicable. Evidence levels and an algorithm to guide personalized therapy were established from literature. Results: 183 genes and 558 potential treatment options for 46 histologies were identified by 01-25-16 (257 evidence items added to civic.genome.wustl.edu). Evidence levels were applied to treatment options for all histological subtypes. More complex phenotypes (e.g. EMT, cancer stem cells) were added to the database tentatively. Thus, a continuously updated clinical algorithm was created, incorporating immune checkpoint status, panel sequencing results, assessment of pre-clinical and clinical evidence levels for molecular features within the clinico-pathological context, leading to a treatment option. Therapy recommendations for the first patients were made using this algorithm. Conclusions: A comprehensive, continuously updated clinical decision algorithm can help improve identification of profiling-based treatment options. Based on collection and analysis of currently known predictive gene aberrations from the literature and major databases, we designed a clinical decision algorithm for precision therapies that we recently implemented within the molecular tumor board at the Charité Comprehensive Cancer Center and whose performance we will systematically evaluate in the future. Comparison to similar efforts by other groups will be useful in order to further develop profiling-based precision therapy of cancer.

Objective To determine the prognostic value of the immunohistochemical evaluation of the multidrug resistance-associated protein 2 (MRP2) expression, together with its subcellular localization in primary fallopian tube carcinomas (PFTCs). Methods The immunohistochemical analysis was performed using samples originating from 70 patients with PFTCs. Results (1) We documented that MRP2 can be localized in the plasma membrane (MRP2c), as well as in the nuclear envelope (MRP2n) of the PFTC cells. (2) Patients with more advanced stage, with progression of the disease and patients who died, showed significantly higher expression of the MRP2n. (3) Univariate and multivariate analyses showed that MRP2n is an unfavorable prognostic factor in PFTCs. (4) The analysis of the classic clinicopathological data revealed that only the FIGO stage had prognostic value, both in the univariate, as well as in multivariate analysis. Conclusions (1) This study suggests that MRP2n is a new disadvantageous prognostic factor in PFTCs and (2) that expression in nuclear envelope can be associated with lower differentiation of cancer cells and their resistance to the cisplatin. (3) We have also confirmed independent prognostic value of FIGO stage in PFTCs.

Estrogen as a potential factor of ovarian carcinogenesis, acts via two nuclear receptors, estrogen receptor alpha (ERα) and estrogen receptor beta (ERβ), but the cellular signal pathways involved are not completely clear so far. In this study we have described the expression of ERα, detected by immunocytochemistry in 11 ovarian carcinoma cell lines and by immunohistochemistry in 43 Federation Internationale des Gyneacologistes et Obstetristes stage III ovarian carcinoma specimens prepared before and after treatment with cisplatin-based schemes. For cisplatin resistance is a major obstacle in the treatment of ovarian carcinoma, analysis of cisplatin sensitivity in 11 ovarian carcinoma cell line was also performed. The strong nuclear ERα expression was only shown in the single A2780P cell line. Expression of ERα in tissue specimens did not reveal any correlations between histopathological parameters (histologic type and grading). We demonstrated a significant association with ERα expression in specimens from primary laparotomies (PL) and cause–specific survival. In the cases terminated by death of the patient, overall immunoreactivity score of ERα expression at PL was significantly lower than in surviving patients. In addition, Kaplan-Meier analysis revealed significantly shorter overall survival time and progression-free time in cases with lower immunoreactivity score of ERα expression at PL. Our findings support the hypothesis that aberrant hormone activity, by way of altered receptor expression, might be an important factor in the malignant transformation of ovarian cancer.

BACKGROUND:Y-Box-binding protein-1 (YB-1) acts as a transcription factor for multiple genes and is linked to DNA replication and repair, cell proliferation and resistance to cytostatic drugs.PATIENTS AND METHODS:The prognostic value of YB-1 expression in primarily untreated malignant non-Hodgkin's lymphomas (NHLs) was examined using immunohistochemistry.RESULTS:Expression of YB-1 was detected in 48 out of 56 NHLs, and the immunohistochemical reaction was localized exclusively in the cytoplasm. Expression of YB-1 did not correlate with clinicopathological variables. Patients with higher YB-1 expression had shorter progression-free survival during the entire period of observation (p=0.0434), as well as in the course of 30 months' observation (p=0.0253). Additionally, in the course of 50 months' observation, patients with higher expression of YB-1 demonstrated a shorter overall survival time (p=0.0383) and a shorter progression-free survival (p=0.0309).CONCLUSION:Elevated YB-1 expression may represent a new unfavorable prognostic factor.

One of the leading causes of chemotherapy failure in non-Hodgkin's lymphomas (NHLs) is multidrug resistance (MDR). MDR can be associated with expression of members of the family of ABC-transporters. Since a correlation between expression of cyclooxygenase-2 (COX-2) and MDR in various cancer cells was described, the expression of COX-2 and the ABC-transporters MDR1/P-glycoprotein (P-gp), MRP1, MRP2 and BCRP was examined in 56 previously non-treated patients by immunohistochemistry. The data show that: i) P-gp is not expressed in non-treated NHLs; ii) MRP2 can be localized in the nuclear membranes of NHL cells; iii) expression of MRP2 in the cytoplasm membrane correlates with clinical response; iv) elevated expression of BCRP is typical for the patients, who did not respond to primary chemotherapy and for cases with shorter progression-free survival time in a 30 months follow-up; and v) there is a strong correlation between COX-2 and MRP1, MRP2 and BCRP. It can be concluded that: i) BCRP may be a crucial factor involved in primary resistance of NHLs, thus it may be useful for prediction of chemotherapeutic treatment and risk of relapse; and ii) since there is strong correlation between COX-2 expression and MDR in NHLs, the application of COX-2 inhibitors may be considered for chemosensitization.

Betulinic acid is a triterpene isolated from the bark of many plants that exhibits cytotoxicity in several cancer cell lines and is capable of inducing apoptosis. In this study, we examined the cytotoxic activity and apoptotic ability of betulinic acid in the drug-sensitive (MeWo) and drug-resistant melanoma MeWo CIS (cisplatin), MeWo ETO (etoposide), MeWo VIN (vinblastin) and MeWo FOTE (fotemusine) cell lines, as well as in the normal melanocyte NHEM-neo cell line. The results show that betulinic acid exhibited significant cytotoxicity on all the cell lines. However, a sulphorhodamine B cell proliferation assay and immunocytochemical analysis of Ki67 expression revealed the strongest cytotoxicity on the normal melanocyte cell line, NHEM-neo. Flow cytometry and immunocytochemical analysis of caspase 3 expression was used to confirm cell death by apoptosis. In conclusion, betulinic acid is a potential candidate for anticancer research, and may also have an application in the cosmetics industry.

Decreased expression of p16 may result from hypermethylation of the promoter or from deletion of the gene. It can lead to intensified proliferation of neoplastic cells and to cytostatic drug resistance. The study was aimed at the examination of prognostic value of p16 expression in relation to Ki67 and caspase-3 in ovarian cancers using immunohistochemistry. The immunohistochemical studies were performed on 73 paraffin-embedded samples of ovarian cancers from 43 patients and samples from 6 healthy ovaries. We have used monoclonal antibodies against p16. ABC method and DAB were used for antigens visualisation. The intensity of the immunohistochemical reactions was appraised using the semi-quantitative IRS scale. In healthy ovaries we have shown strong reaction in the nuclei of surface epithelium. In the case of studied ovarian cancers, the reaction of a nuclear and cytoplasmic localization was obtained. The mean overall immunoreactivity score of nuclear p16 expression amounted to 5.30+/-3.44 SD in primary laparotomy material and 6.61+/-4.34 SD in secondary cytoreduction material. Statistical analysis demonstrated that lower p16 expression was typical of the younger patients and the patients who died. Kaplan-Meier's analysis proved that lower expression of p16 was characteristic of cases with shorter overall survival. In the present study we have demonstrated that lowered p16 expression represented an unfavourable prognostic index in ovarian cancer. Lowered p16 expression was also typical for chemotherapy-resistant ceases (cases of lower caspase-3 and higher Ki67 at secondary cytoreduction expression).

An obstacle in chemotherapy of ovarian cancer is the development of drug resistance. Taxol (paclitaxel)-resistance-associated gene-3 (TRAG-3/CSAG2) was found to be overexpressed in a paclitaxel-resistant ovarian carcinoma cell line. However, clinical impact of TRAG-3 in ovarian carcinoma has not been demonstrated previously. For demonstration of potential clinical impact of TRAG-3, immunohistochemistry was applied to determine TRAG-3 protein expression in specimens obtained from ovarian carcinoma patients (n=37) who received a paclitaxel-based chemotherapy at two different time points, initial laparotomy before chemotherapy, and secondary cytoreduction after chemotherapy. The TRAG-3-specific immunohistochemical staining was correlated with clinical outcome. In ovarian carcinoma specimens obtained at the initial laparotomy, an advantage in overall (P < 0.001) and progression-free (P = 0.003) survival for patients with weak TRAG-3 expression could be demonstrated. Tumor specimens excised at secondary cytoreduction procedure were not predictive for clinical outcome. In summary, TRAG-3 was found to be a prognostic factor for the prediction of clinical outcome after the application of paclitaxel-based chemotherapy.

BACKGROUND Numerous experimental studies have described the capacity of myofibroblasts to stimulate mammary cancer cells in a paracrine manner. Until now, the prognostic significance of myofibroblasts present in breast cancer has not been examined. PATIENTS AND METHODS In paraffin sections, originating from 45 patients with primary invasive breast cancer, immunohistochemical reactions were performed using antibodies directed against smooth muscle actin, Ki-67, VEGF, bFGF and UPA. RESULTS The cases with higher content of myofibroblasts in the tumour tissue manifested higher grade, more pronounced expression of Ki-67, VEGF and bFGF and shorter overall survival and relapse-free survival. CONCLUSION The present study for the first time documents the unfavourable prognostic significance of myofibroblasts in tissues of invasive ductal mammary carcinomas.

HistopathologyVolume 51, Issue 1 p. 125-127 Unfavourable prognostic significance of S100P expression in ovarian cancers P Surowiak, P Surowiak Institute of Pathology, Charité Campus Mitte, Berlin, Germany Department of Histology and Embryology, University School of Medicine Lower Silesian Centre of Oncology, WrocławSearch for more papers by this authorA Maciejczyk, A Maciejczyk Lower Silesian Centre of Oncology, WrocławSearch for more papers by this authorV Materna, V Materna Institute of Pathology, Charité Campus Mitte, Berlin, GermanySearch for more papers by this authorM Drag-Zalesińska, M Drag-Zalesińska Department of Histology and Embryology, University School of MedicineSearch for more papers by this authorA Wojnar, A Wojnar Lower Silesian Centre of Oncology, WrocławSearch for more papers by this authorM Pudelko, M Pudelko Lower Silesian Centre of Oncology, WrocławSearch for more papers by this authorW Kędzia, W Kędzia Department of Obstetrics and GynaecologySearch for more papers by this authorM Spaczyński, M Spaczyński Department of Obstetrics and GynaecologySearch for more papers by this authorM Dietel, M Dietel Institute of Pathology, Charité Campus Mitte, Berlin, GermanySearch for more papers by this authorM Zabel, M Zabel Department of Histology and Embryology, University School of Medicine Department of Histology and Embryology, University School of Medicine, Poznań, PolandSearch for more papers by this authorH Lage, H Lage Institute of Pathology, Charité Campus Mitte, Berlin, GermanySearch for more papers by this author P Surowiak, P Surowiak Institute of Pathology, Charité Campus Mitte, Berlin, Germany Department of Histology and Embryology, University School of Medicine Lower Silesian Centre of Oncology, WrocławSearch for more papers by this authorA Maciejczyk, A Maciejczyk Lower Silesian Centre of Oncology, WrocławSearch for more papers by this authorV Materna, V Materna Institute of Pathology, Charité Campus Mitte, Berlin, GermanySearch for more papers by this authorM Drag-Zalesińska, M Drag-Zalesińska Department of Histology and Embryology, University School of MedicineSearch for more papers by this authorA Wojnar, A Wojnar Lower Silesian Centre of Oncology, WrocławSearch for more papers by this authorM Pudelko, M Pudelko Lower Silesian Centre of Oncology, WrocławSearch for more papers by this authorW Kędzia, W Kędzia Department of Obstetrics and GynaecologySearch for more papers by this authorM Spaczyński, M Spaczyński Department of Obstetrics and GynaecologySearch for more papers by this authorM Dietel, M Dietel Institute of Pathology, Charité Campus Mitte, Berlin, GermanySearch for more papers by this authorM Zabel, M Zabel Department of Histology and Embryology, University School of Medicine Department of Histology and Embryology, University School of Medicine, Poznań, PolandSearch for more papers by this authorH Lage, H Lage Institute of Pathology, Charité Campus Mitte, Berlin, GermanySearch for more papers by this author First published: 31 May 2007 https://doi.org/10.1111/j.1365-2559.2007.02714.xCitations: 14Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL References 1 Györffy B, Surowiak P, Kiesslich O et al. Resistance prediction profile for eleven anticancer agents at clinical concentrations based on the gene expression pattern of thirty cell lines. Int. J. Cancer 2006; 118; 1699– 1712. 2 Jiang F, Shults K, Flye L et al. S100P is selectively upregulated in tumor cell lines challenged with DNA cross-linking agents. Leuk. Res. 2005; 10; 1181– 1190. 3 Bertram J, Palfner K, Hiddemann W, Kneba M. Elevated expression of S100P, CAPL and MAGE 3 in doxorubicin-resistant cell lines: comparison of mRNA differential display reverse transcription-polymerase chain reaction and subtractive suppressive hybridization for the analysis of differential gene expression. Anicancer Drugs 1998; 9; 311– 317. 4 Arumugam T, Simeone DM, Schmidt AM, Logsdon CD. S100P stimulates cell proliferation and survival via receptor for activated glycation end products (RAGE). J. Biol. Chem. 2004; 279; 5059– 5065. 5 Surowiak P, Materna V, Denkert C et al. Significance of cyclooxygenase 2 and MDR1/P-glycoprotein coexpression in ovarian cancers. Cancer Lett. 2006; 235; 272– 280. 6 Remmele W, Stegner HE. [Recommendation for uniform definition of an immunoreactive score (IRS) for immunohistochemical estrogen receptor detection (ER-ICA) in breast cancer tissue][in German]. Pathologe 1987; 8; 138– 140. Citing Literature Volume51, Issue1July 2007Pages 125-127 ReferencesRelatedInformation

A major obstacle in treatment of ovarian cancer is intrinsic or acquired drug resistance causing failure of chemotherapy followed by a poor clinical outcome. Drug resistance of ovarian carcinoma can be caused by dysregulation of cellular factors involved in regulation of apoptosis and DNA repair pathways. In this study, 73 ovarian carcinoma specimens obtained before and after chemotherapy were analysed by immunohistochemistry for expression of seven proteins playing an important role in regulation of DNA mismatch repair and apoptosis. The prognostic significance of these proteins in the meaning of overall and progression-free survival was evaluated in univariate and multivariate analysis. Bcl-xL, hMSH2, caspase-3, p21 and p53 displayed prognostic importance in univariate analysis. Furthermore, it was demonstrated that caspase-3 and p21 were also independent prognostic markers for both, overall and progression-free survival. In conclusion, these data indicate that analysis of proteins involved in DNA mismatch repair and apoptosis can be useful for prediction of clinical outcome in ovarian carcinoma patients.

Elevated metallothionein (MT) expression in ovarian cancers treated with cisplatin-based schemes represents an unfavorable prognostic index. MT expression is significantly higher in tumor samples obtained after chemotherapy. The present study aimed at examining MT expression in ovarian carcinoma cells sensitive (A2780) or resistant (A2780RCIS) against platinum drug treatment as well as examining effects of exposure to cisplatin on MT expression. Subcellular expression of MT was evaluated also in samples originating from 73 ovarian tumors. Cisplatin-resistant A2780RCIS cells were exposed to increasing cisplatin concentrations, and the subcellular expression of MT was determined by immunocytochemistry. The studies demonstrated that cisplatin-resistant A2780RCIS cells exposed to cisplatin typically manifested a nuclear MT expression. The study demonstrated also that exposure to cisplatin was paralleled by growing MT expression in cell nuclei. The nuclear expression of MT was also found to be specific for ovarian cancers of poor clinical outcome. No relationship could be demonstrated between cytoplasmic expression of MT and clinical variables. Nuclear MT expression is induced by cisplatin and seems to protect DNA in the cells from toxic effects of the drug.

In the chemotherapeutic treatment of patients with disseminated neoplasms, multidrug resistance (MDR) is a major obstacle. ABCG2 (BCRP/MXR), a member of the superfamily of adenosine triphosphate-binding cassette (ABC) transporters, was demonstrated to be associated with "atypical" forms of multidrug-resistant phenotypes of cancer cells. To overcome the ABCG2-depending MDR, two specific anti-ABCG2 small interfering RNAs (siRNAs) were designed for transient triggering of the gene-silencing RNA interference (RNAi) pathway in the human gastric carcinoma cell line EPG85-257RNOV, exhibiting an atypical MDR phenotype. Because both siRNAs showed biological activity, for stable inhibition of ABCG2 corresponding short hairpin RNA (shRNA) expression vectors were constructed. By treatment of EPG85-257RNOV cells with these constructs, expression of the targeted ABCG2-encoding mRNA and transport protein was inhibited completely. Furthermore, anti-ABCG2 shRNA-treated cells increased cellular drug accumulation to the same level measured in drug-sensitive parental cells. These effects were accompanied by complete reversal of the drug-resistant phenotype. Thus, the data indicate that siRNA- and shRNA-mediated RNAi-based gene therapy may be applicable in preventing and reversing ABCG2-depending atypical MDR.

BACKGROUND:In breast cancer, the expression of CD24 represents a poorly recognised unfavourable prognostic factor. CD24 has been described to be potentially down-regulated by estrogen receptor alpha (ER). The present study was aimed at examining the predictive value of CD24 expression in tamoxifen-treated breast cancer cases.MATERIALS AND METHODS:Sixty patients with primary invasive ductal breast cancers with post-operative tamoxifen treatment were enrolled in the study. Immmunohistochemical reactions were performed using monoclonal antibodies directed against CD24 and ER.RESULTS:Cases demonstrating cytoplasmic-membranous expression of CD24 (CD24c-m) proved to be characterised by a significantly lower expression of ER as compared to CD24c-m-negative cases. A multivariate progression analysis based on the Cox proportional hazard model demonstrated that CD24c-m expression is an independent prognostic factor for poor overall survival.CONCLUSION:The data from the present study suggested that CD24c-m expression is specific for tamoxifen-resistant breast cancer cases. CD24 should be subjected to comprehensive studies as a marker of resistance to tamoxifen treatment.