Masatoshi Mukai

Infiltration of tumour‐associated macrophages in prostate biopsy specimens is predictive of disease progression after hormonal therapy for prostate cancer

Study Type – Prognostic (case series)

Excision Repair Cross-Complementing Group 1 May Predict the Efficacy of Chemoradiation Therapy for Muscle-Invasive Bladder Cancer

Purpose: Chemoradiation therapy (CRT) is now widely recognized as bladder-preserving therapy for muscle-invasive bladder cancer (MIBC). However, some patients who fail CRT may miss the chance to be cured by cystectomy. Therefore, it is important to select patients with MIBC who are expected to have a good response to CRT. Several reports indicate that the excision repair cross-complementing group 1 (ERCC1) gene is associated with resistance to cisplatin and radiation therapy. In this study, we examined the correlation between ERCC1 and CRT in vitro and in vivo in bladder cancer. Experimental Design: Bladder cancer cell lines T24, 5637, Cl8-2 (multidrug-resistant subline of T24), and CDDP10-3 (cisplatin-resistant subline of T24) were used for in vitro assays to measure ERCC1 expression level and growth inhibition with cisplatin or ionizing radiation (IR). We then examined by immunohistochemistry that whether ERCC1 nuclear staining correlates with the efficacy of CRT using cisplatin in 22 patients with MIBC. Results: Cl8-2 cells expressed ERCC1 mRNA 5.96-fold higher than did T24. Cl8-2 and CDDP10-3 were more resistant to cisplatin or IR than was T24. Resistance to IR, but not to cisplatin, was removed by suppressing ERCC1 using siRNA in both Cl8-2 and CDDP10-3 cells. In immunohistochemistry with ERCC1, 6 of 8 positive cases did not have complete response to CRT, whereas 12 of 14 negative cases had complete response. Sensitivity and specificity were 75% and 85.7%, respectively (P = 0.008). Conclusion: Although further study is needed, ERCC1 expression level may predict the efficacy of CRT for MIBC. Clin Cancer Res; 17(8); 2561–9. ©2010 AACR.

SERPINE2 is a possible candidate promotor for lymph node metastasis in testicular cancer

Testicular germ cell tumors (TGCTs) commonly metastasize to the lymph node or lung. However, it remains unclear which genes are associated with TGCT metastasis. The aim of this study was to identify gene(s) that promoted human TGCT metastasis. We intraperitoneally administered conditioned medium (CM) from JKT-1, a cell-line from a human testicular seminoma, or JKT-HM, a JKT-1 cell sub-line with high metastatic potential, into mice with JKT-1 xenografts. Administration of CM from JKT-HM significantly promoted lymph node metastasis. A cDNA microarray analysis showed that JKT-HM cells highly expressed the Serpine peptidase inhibitor, clade E, member 2 (SERPINE2), which encodes a secreted protein. Administration of CM from SERPINE2-silenced JKT-HM cells inhibited lymph node metastasis in the xenograft model, compared with administration of CM from JKT-HM cells. There was no significant difference in xenograft volume. Moreover, administration of CM from SERPINE2-over-expressing JKT-1 was likely to promote lymph node metastasis in the xenograft model. There was no difference in the in vitro proliferation or migration of JKT-1 cells cultured with CM from JKT-HM cells, compared to that with CM from JKT-1. There was no promotion of proliferation or lymphangiogenesis in the xenografts, as measured by Ki-67 and LYVE-1 immunohistochemistry, respectively. Although we could not clarify how SERPINE2 promoted lymph node metastasis, it may be a promoter in the development of lymph node metastasis in the human seminoma cells in a mouse xenograft model.

Brain metastases from testicular germ cell tumors: A retrospective analysis

Objectives:  To review our series of testicular germ cell tumors with brain metastases and to establish an optimal treatment strategy for them.

Retrospective Analysis of an Oral Combination of Dexamethasone, Uracil plus Tegafur and Cyclophosphamide for Hormone-refractory Prostate Cancer

OBJECTIVE To evaluate the clinical utility of an oral combination of dexamethasone, uracil plus tegafur and cyclophosphamide as a treatment for patients with hormone-refractory prostate cancer. METHODS Fifty-seven patients with hormone-refractory prostate cancer were treated with an oral administration of dexamethasone (1.0 mg/day), uracil plus tegafur (400 mg/day) and cyclophosphamide (100 mg/day). The median patient age was 71 years. Sixteen patients had symptomatic bone metastasis, 31 had asymptomatic bone metastasis and 8 showed lymph node metastasis. Eight patients presented with only biochemical progression as evaluated by serum prostate-specific antigen levels. RESULTS Thirty-six (63%) of 57 patients demonstrated a ≥50% decline in serum prostate-specific antigen levels. The median time to prostate-specific antigen progression was 7.2 months. In patients with a prostate-specific antigen decline of ≥50%, the median time to progression was 13.3 months. With respect to pre-treatment markers, the duration of response to initial hormonal treatment was associated with the time to prostate-specific antigen progression. In 11 of 16 (69%) patients who complained of bone pain, the pain improved and became stable in 5 of those patients (31%). Most adverse events were mild and only three (5%) patients showed neutropenia of Grade 3 or higher. CONCLUSIONS The combination of dexamethasone, uracil plus tegafur and cyclophosphamide is an effective and well tolerated regimen for hormone-refractory prostate cancer. To evaluate the survival benefits, further randomized studies are required.

Decreased infiltration of macrophage scavenger receptor-positive cells in initial negative biopsy specimens is correlated with positive repeat biopsies of the prostate

Macrophage scavenger receptor (MSR)‐positive inflammatory cells and tumor‐associated macrophages (TAMs) have been reported to regulate the growth of various cancers. In this study, the infiltration of MSR‐positive cells and TAMs was analyzed to predict the outcome of repeat biopsy in men diagnosed as having no malignancy at the first prostate biopsy. Repeat biopsy of the prostate was carried out in 92 patients who were diagnosed as having no malignancy at the first biopsy. Of these, 30 patients (32.6%) were positive for prostate cancer at the repeat biopsy. Tumor‐associated macrophages and MSR‐positive cells were immunohistochemically stained with mAbs CD68 and CD204, respectively. Six ocular measuring fields were chosen randomly under a microscope at ×400 power in the initial negative biopsy specimens, and the mean TAM and MSR counts for each case were determined. No difference in TAM count was found between the cases with or without prostate cancer. By contrast, the MSR count in patients with cancer was significantly lower than that in patients without cancer at the repeat biopsy (P < 0.001). Logistic regression analysis indicated that the MSR count at first biopsy is a significantly better predictive factor for positive repeat biopsy than PSA velocity, interval between first and repeat biopsies, or TAM count. Decreased infiltration of MSR‐positive cells in negative first biopsy specimens was correlated with positive findings in the repeat biopsy. The MSR count might be a good indicator for avoiding unnecessary repeat biopsies. (Cancer Sci 2010)

Tumor multiplicity is an independent prognostic factor of non-muscle-invasive high-grade (T1G3) bladder cancer.

OBJECTIVE Non-muscle-invasive high-grade (T1G3) bladder cancers have high potential for progression. The objective of this study is to clarify the clinicopathological factors affecting the outcome of T1G3 bladder cancer. METHODS We retrospectively reviewed 60 cases of T1G3 bladder cancer between 1994 and 2006. The correlations of both intravesical recurrence and progression with prognostic factors, such as T stage, history of bladder cancer, multiplicity, concomitant carcinoma in situ, tumor size, intravesical instillation of bacillus Calmette-Guérin and intravesical chemotherapy, were evaluated by multivariate analysis with the Cox proportional hazards model. RESULTS Median follow-up period was 52 months (4-105 months). Thirty-seven cases of intravesical recurrence (61.7%) were observed during follow-up. Two- and 5-year recurrence-free survival rates were 44.1% and 36.1%, respectively. Tumor multiplicity and instillation of bacillus Calmette-Guérin were significantly correlated with intravesical recurrence on multivariate analysis. Ten cases of progression (16.7%) were observed during the follow-up period. Two- and 5-year progression-free survival rates were 87.7% and 83.4%, respectively. Only tumor multiplicity was significantly correlated with progression on multivariate analysis. CONCLUSIONS T1G3 cancers with multiple lesions showed high risks of intravesical recurrence and progression. Although bacillus Calmette-Guérin instillation reduced the risk of intravesical recurrence, no effect was observed on disease progression.

1998 IMPACT OF HYPONATREMIA ON SURVIVAL OF PATIENTS WITH METASTATIC RENAL CELL CARCINOMA TREATED WITH MOLECULAR TARGETED THERAPY

Atsunari Kawashima*, Hitoshi Takayama, Suita, Japan; Yasuyuki Arai, Osaka, Japan; Mikio Nin, Sakai, Japan; Go Tanigawa, Yutaka Yasunaga, Osaka, Japan; Masatoshi Mukai, Toyonaka, Japan; Hironori Nomura, Daizo Oka, Toshiaki Yoshioka, Osaka, Japan; Satoko Fukuda, Ikeda, Japan; Kenji Nishimura, Nishinomiya, Japan; Nobukazu Murosaki, Itami, Japan; Minoru Koga, Minoh, Japan; Yasuyuki Kojima, Suita, Japan; Miyaji Kyakuno, Osaka, Japan; Takahiro Yoshida, Koji Hatano, Mototaka Sato, Motohide Uemura, Yasutomo Nakai, Suita, Japan; Kazuo Nishimura, Osaka, Japan; Akira Tsujimura, Norio Nonomura, Suita, Japan

Abstract 2475: Excision repair cross complementing group 1 (ERCC1) predicts the efficacy of chemoradiotherapy for invasive bladder cancer

Purpose: Chemoradiation therapy (CRT) is now widely recognized as bladder-preserving therapy for muscle-invasive bladder cancer (MIBC). However, some patients who fail CRT may miss the chance to be cured by cystectomy. Therefore, it is important to select patients with MIBC who are expected to have a good response to CRT. Several reports indicate that the excision repair cross-complementing group 1 (ERCC1) gene is associated with resistance to cisplatin and radiation therapy. In this study, we examined the correlation between ERCC1 and CRT in vitro and in vivo in bladder cancer. Experimental Design: Bladder cancer cell lines T24, 5637, Cl8-2 (multi-drug-resistant subline of T24), and CDDP10-3 (cisplatin-resistant subline of T24) were used for in vitro assays to measure ERCC1 expression level and growth inhibition with cisplatin or irradiation (IR). To clarify the association between ERCC1 and cisplatin resistance in bladder cancer cells, we knocked down ERCC1 in Cl8-2 and CDDDP10-3 with siRNA (C18-2ΔERCC1 and CDDP10-3ΔERCC1). To further prove the cause of the radiation sensitivity of ERCC1 knockdown cells, we measured the phosphorylated histone variant H2A.X, a marker of DNA damage. In the clinical study, we then examined by immunohistochemistry whether ERCC1 nuclear staining correlates with the efficacy of CRT using cisplatin in 22 patients with MIBC. Results: Cl8-2 cells expressed ERCC1 mRNA 5.96-fold higher than did T24. Cl8-2 and CDDP10-3 were more resistant to cisplatin or IR than was T24. Our ERCC1 knockdown experiments showed that there was statistical difference in the resistance to IR exposure not cisplatin compared with C18-2CTL and CDDP10-3CTL. C18-2ΔERCC1 and CDDP10-3ΔERCC1 recovered more slowly in terms of the number of phospho-H2A.X foci than did C18-2cont and CDDP10-3cont, suggesting continued accumulation or persistence of DSBs. In immunohistochemistry with ERCC1, six of eight positive cases did not have complete response to CRT, whereas 12 of 14 negative cases had complete response. Sensitivity and specificity were 75% and 85.7%, respectively (p = 0.008). Conclusion: Our results suggest that in some bladder cancer cells, ERCC1 expression correlates with IR resistance but not with cisplatin resistance. Moreover, the lack of ERCC1 expression correlated well with the efficacy of CRT, and especially with that of IR, in our clinical study. Although further study is needed, ERCC1 expression level may predict the efficacy of CRT for MIBC. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2475. doi:10.1158/1538-7445.AM2011-2475

Abstract 3186: Increased number of mast cells infiltrating in non-malignant biopsy is associated with detection of prostate cancer

Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL INTRODUCTION AND OBJECTIVE: Prostate cancer (PCA) is one of the most common malignancies. Prostate-specific antigen (PSA) is currently the most useful serum tumor marker for detecting PCA. However, because of the low specificity of PSA, many patients undergo unnecessary needle biopsies of the prostate. The discovery of new predictors is essential to establish appropriate repeat biopsies. Tumor cells are surrounded by inflammatory cells such as macrophages, lymphocytes, neutrophils and mast cells(MCs). Inflammatory cell infiltrates may contribute to tumor progression by producing angiogenic factors. MCs are potent stimulators of angiogenesis, and condition cancer growth and development of the metastasis. In the present study, infiltration of MCs in non-malignant biopsy specimens was evaluated as a possible predictive value for PCA. METHODS: These reviews were evaluated for 125 patients ranging from 51 to 82 years (median, 69 years), who received repeat needle biopsies of the prostate and were diagnosed as non-malignant during first biopsy. Of these, 40 (32.0%) were positive for cancer during second or third biopsy. MCs was immunohistochemically labeled using a mast cell tryptase monoclonal antibody. For systematic counting, 6 ocular measuring fields were randomly chosen under a microscope at a power of×400 within first negative biopsy specimens. RESULTS: Serum PSA levels measured by the immunoenzymatic assay from 1.70 to 43.18 ng/ml (median, 8.30 ng/ml) at first biopsy. Of these, 30 (32.0%) were positive for cancer during second, third or fourth biopsy. Serum PSA levels ranged from 1.90 to 52.68 ng/ml (median, 10.01 ng/ml) at the time of biopsy of detection of PCA. The period between the date of first biopsy and diagnosis of PCA ranged from 4.1 to 70.1 months (mean, 25.7 months).There were no significant differences in PSA at first biopsy, age and interval of previous and subsequent biopsies, between negative and positive cases for cancer. However, in PSA at first and repeat biopsy, there were significant differences between patients with or without cancer (P = 0.014, 0.002). MCs were observed in all specimens tested. Median count of MCs in each case was 11. MCs count in patients with cancer was higher than in those without cancer (P < 0.001). Frequency of PCA in high MCs group (≥ 11) (54.5%) was much higher than that in low MCs group (< 11) (12.5%) (P < 0.001). Frequency of PCA in high PSA group (≥ 9.0) (43.8%) was much higher than that in low PSA group (<9.0) (20.5%) (P = 0.015). CONCLUSION: Increased number of Mast cells infiltrating in non-malignant biopsy Is Associated with Detection of PCA. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3186. doi:10.1158/1538-7445.AM2011-3186