Genotype-Dependent Efficacy of a Dual PI3K/mTOR Inhibitor, NVP-BEZ235, and an mTOR Inhibitor, RAD001, in Endometrial Carcinomas

The PI3K (phosphatidylinositol-3-kinase)/mTOR (mammalian target of rapamycin) pathway is frequently activated in endometrial cancer through various PI3K/AKT-activating genetic alterations. We examined the antitumor effect of NVP-BEZ235—a dual PI3K/mTOR inhibitor—and RAD001—an mTOR inhibitor—in 13 endometrial cancer cell lines, all of which possess one or more alterations in PTEN, PIK3CA, and K-Ras. We also combined these compounds with a MAPK pathway inhibitor (PD98059 or UO126) in cell lines with K-Ras alterations (mutations or amplification). PTEN mutant cell lines without K-Ras alterations (n = 9) were more sensitive to both RAD001 and NVP-BEZ235 than were cell lines with K-Ras alterations (n = 4). Dose-dependent growth suppression was more drastically induced by NVP-BEZ235 than by RAD001 in the sensitive cell lines. G1 arrest was induced by NVP-BEZ235 in a dose-dependent manner. We observed in vivo antitumor activity of both RAD001 and NVP-BEZ235 in nude mice. The presence of a MEK inhibitor, PD98059 or UO126, sensitized the K-Ras mutant cells to NVP-BEZ235. Robust growth suppression by NVP-BEZ235 suggests that a dual PI3K/mTOR inhibitor is a promising therapeutic for endometrial carcinomas. Our data suggest that mutational statuses of PTEN and K-Ras might be useful predictors of sensitivity to NVP-BEZ235 in certain endometrial carcinomas

Long-term monitoring of sediment runoff for an active sediment control in Joganji River

There were huge sediment yielding and deposition due to debris flows by breaking natural landslide dams which were formed by earthquake in 1858 at upstream reach of Joganji River. Sediment transportation is still active by debris flow and flow with bedload due to rainfall, though a lot of erosion control dams have been constructed. Continuously measuring sediment runoff for long term along a main river is necessary to evaluate the propagation of sediment after the huge events for sediment management in the basin using well hydrological information. Appropriate tools are selected and applied to monitoring in the area managed by Tateyama Mountain Area Sabo Office along Joganji River, using a Reid-type bedload slot sampler, robust-type hydrophone and velocity meter on the bed for bedload and turbidity meter for washload. Monitored data is concentratedly collected at the office to apply risk management for sediment movement due to heavy rainfall and so on. Several typical data and problems to solved were shown because it passed around twenty years since sediment monitoring started, and those are reported in present study

Cytological Analysis for Human Papillomavirus DNAs in Cervical Intraepithelial Neoplasia by In situ Hybridization

Human papillomavirus (HPV) type 16 and 18 DNAs are reported to be associated with uterine cervical cancer. In order to investigate the relationship between the presence of HPV DNA and cervical intraepithelial neoplasia (CIN), we attempted the cytological detection of HPV DNAs in uterine cervical smear samples. The samples included those of severe dysplasia and carcinoma in situ (CIS). They were analysed by DNA-DNA in situ hybridization using biotinylated HPV DNA probes.　　 The results of in situ hybridization analysis revealed that HPV sequences were present in the nuclei of cells with koilocytotic atypia. When probed for HPV type 6, 11, 16 and 18, the nuclei of dysplastic cells and cancer cells were positive for HPV type 16 and 18 DNA. Out of 26 CIN cases, 17 contained HPV type 16 DNA and 5 contained HPV type 18 DNA.　　 We suggest that cytological analysis for HPV sequences by the in situ hybridization technique might provide a molecular diagnosis for assessment in uterine cervical intraepithelial neoplasia.This work was supported in part by a Grant-in-Aid from the Ministry of Education, Science and Culture of Japan (No.01579833, No.04671004)

Clinical Evaluation of Human Granulocyte Colony-stimulating Factor in Chemotherapy for Ovarian Cancer

The clinical evaluation of human granulocyte colony-stimulating factor (G-CSF) in 38 patients treated with chemotherapy for ovarian cancer stage III was investigated among 3 groups. G-CSF was not given in group A (19 courses), was administered from day 5 of chemotherapy in group B (53 courses), and was given when the WBC count decreased to below 2,000/mm3 in group C (29 courses).　　 The time to nadir was significantly shorter in group B compared with groups A and C and revealed 10 days for the WBC count and 11 days for the neutrophil count (p<0.01), with mean nadir values of 2,896/mm3 and 982/mm3 respectively, and so the count of WBC and neutrophil have been kept during the course. The effect of G-CSF was not modified by age, body weight or the number of chemotherapy courses in groups B and C. These results demonstrate that early treatment with G-CSF may allow increased intensity of chemotherapy by using greater doses or by shortening of the interval between cycles

Long-term monitoring of sediment runoff for an active sediment control in Joganji River

There were huge sediment yielding and deposition due to debris flows by breaking natural landslide dams which were formed by earthquake in 1858 at upstream reach of Joganji River. Sediment transportation is still active by debris flow and flow with bedload due to rainfall, though a lot of erosion control dams have been constructed. Continuously measuring sediment runoff for long term along a main river is necessary to evaluate the propagation of sediment after the huge events for sediment management in the basin using well hydrological information. Appropriate tools are selected and applied to monitoring in the area managed by Tateyama Mountain Area Sabo Office along Joganji River, using a Reid-type bedload slot sampler, robust-type hydrophone and velocity meter on the bed for bedload and turbidity meter for washload. Monitored data is concentratedly collected at the office to apply risk management for sediment movement due to heavy rainfall and so on. Several typical data and problems to solved were shown because it passed around twenty years since sediment monitoring started, and those are reported in present study

Cyclin D1 harboring the T286I mutation promotes oncogenic activation in endometrial cancer

金沢大学医薬保健研究域医学系Cyclin D1 is an important regulator of cell cycle progression. Phosphorylation of cyclin D1 at Thr286 by GSK3β triggers its nuclear export and cytoplasmic proteolysis via the 26S proteasome. Cyclin D1 overexpression is a common event in various types of human cancers; however, reports of mutations are extremely rare. We analyzed mutations of the cyclin D1 gene, CCND1, in 88 endometrial cancer tissue specimens and detected mutations in 2 cases (2.3%). Both were unreported mutations with substitution of threonine to isoleucine at codon 286 (T286I). These two tumors harbored coexisting mutations in K-ras, PIK3CA and/or PTEN and showed accumulation of cyclin D1 in the nucleus by immunohistochemistry. Furthermore, we analyzed the functions of mutant cyclin D1 (T286I) by luciferase assays, immunofluorescence, western blotting and clonogenic cell survival assays in HEK-293T cells. We found that exogenous mutant cyclin D1 (T286I) accumulated in the nuclei in HEK-293T cells, and that it inhibited the expression of pRb. Additionally, the number of colonies was increased by introduction of mutant cyclin D1 (T286I) compared to that of wild-type cyclin D1. In conclusion, we identified an unreported CCND1 mutation (T286I) in two endometrial cancers and revealed that the mutation was functional for inducing cell proliferation in human cells

Integrated copy number and expression analysis identifies profiles of whole-arm chromosomal alterations and subgroups with favorable outcome in ovarian clear cell carcinomas.

Ovarian clear cell carcinoma (CCC) is generally associated with chemoresistance and poor clinical outcome, even with early diagnosis; whereas high-grade serous carcinomas (SCs) and endometrioid carcinomas (ECs) are commonly chemosensitive at advanced stages. Although an integrated genomic analysis of SC has been performed, conclusive views on copy number and expression profiles for CCC are still limited. In this study, we performed single nucleotide polymorphism analysis with 57 epithelial ovarian cancers (31 CCCs, 14 SCs, and 12 ECs) and microarray expression analysis with 55 cancers (25 CCCs, 16 SCs, and 14 ECs). We then evaluated PIK3CA mutations and ARID1A expression in CCCs. SNP array analysis classified 13% of CCCs into a cluster with high frequency and focal range of copy number alterations (CNAs), significantly lower than for SCs (93%, P < 0.01) and ECs (50%, P = 0.017). The ratio of whole-arm to all CNAs was higher in CCCs (46.9%) than SCs (21.7%; P < 0.0001). SCs with loss of heterozygosity (LOH) of BRCA1 (85%) also had LOH of NF1 and TP53, and LOH of BRCA2 (62%) coexisted with LOH of RB1 and TP53. Microarray analysis classified CCCs into three clusters. One cluster (CCC-2, n = 10) showed more favorable prognosis than the CCC-1 and CCC-3 clusters (P = 0.041). Coexistent alterations of PIK3CA and ARID1A were more common in CCC-1 and CCC-3 (7/11, 64%) than in CCC-2 (0/10, 0%; P < 0.01). Being in cluster CCC-2 was an independent favorable prognostic factor in CCC. In conclusion, CCC was characterized by a high ratio of whole-arm CNAs; whereas CNAs in SC were mainly focal, but preferentially caused LOH of well-known tumor suppressor genes. As such, expression profiles might be useful for sub-classification of CCC, and might provide useful information on prognosis