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19. Photoreceptor apoptosis induced by a single systemic administration of N-methyl-N-nitrosourea in the rat retina

Author

Nakajima, M., Yuge, K., Senzaki, H., Shikata, N., Miki, H., Uyama, M., and Tsubura, A.

Subjects
Cell Nucleus, Retinal Degeneration, Apoptosis, Cell Count, Methylnitrosourea, Retina, Rats, Rats, Sprague-Dawley, Microscopy, Electron, Proliferating Cell Nuclear Antigen, Rats, Inbred BN, Animals, Vimentin, Female, Photoreceptor Cells, sense organs, Pigment Epithelium of Eye, Injections, Intraperitoneal, Research Article
Abstract

Retinal degeneration was induced by a single intraperitoneal injection of N-methyl-N-nitrosourea in female Sprague-Dawley albino rats at 50 days of age by two dose regimens, which were observed sequentially at 24, 48, and 72 hours and 7, 21, and 35 days after the treatment. After a dose of 75 mg/kg, methylnitrosourea evoked progressive retinal degeneration in all treated rats whereas a dose of 50 mg/kg was less effective. The 75-mg/kg-treated rats showed selective destruction of the photoreceptor cells by an apoptotic mechanism, as confirmed morphologically and by the terminal dUTP nick end labeling method. Apoptosis had already started at 24 hours after the treatment and was completed by day 7. During the photoreceptor degeneration, proliferation of glial fibrillary acidic protein and vimentin-positive Müller cells as detected by proliferating cell nuclear antigen labeling appeared at 48 hours and was prominent 72 hours after the treatment, and macrophage infiltration within the retina as recognized by ED1 positivity was maximal 7 and 21 days after the treatment. Retinal degeneration was also induced in female Brown-Norway colored rats in a similar dose-dependent manner. Pigment epithelium was discontinuous above Bruch's membrane, and migration of the swollen pigment epithelium toward the inner nuclear layer was seen 7 days after the treatment. Therefore, as also confirmed electron microscopically, the most striking change was the destruction of photoreceptor cells by the apoptotic process, followed by Müller cell proliferation, pigment epithelium migration, and macrophage infiltration for cell debris phagocytosis, resulting in a thin remnant of retina with attenuated inner nuclear cells in direct contact with Bruch's membrane or with the pigment epithelium and/or with the Müller cells 35 days after the treatment.

Published
1996

26. Abstracts

Author

Zhou, Xinhua, primary, Kudo, Akihiko, additional, Kawakami, Hayato, additional, Hirano, Hiroshi, additional, FAYED, M.H., additional, MAKITA, T., additional, SUZAKI, Etsuko, additional, KATAOKA, Katsuko, additional, Katsumata, Osamu, additional, Fujimoto, Kazushi, additional, Yamashina, Shohei, additional, USUDA, Nobuteru, additional, JOHKURA, Kohhei, additional, SUGANUMA, Tatsuo, additional, SAWAGUCHI, Akira, additional, NAGAIKE, Ryoko, additional, KAWANO, Jun-ichi, additional, OINUMA, Tsutomu, additional, Izumi, Shin-ichi, additional, Iwamoto, Michiko, additional, Shin, Masashi, additional, Nakano, Paul K., additional, Ueda, Tadashi, additional, Ishikawa, Yoshimaro, additional, Kubo, Eri, additional, Miyoshi, Norio, additional, Fukuda, Masaru, additional, Akagi, Yoshio, additional, Miki, H., additional, Nakajima, M., additional, Yuge, K., additional, Taomoto, M., additional, Tsubura, A., additional, Shikata, N., additional, Senzaki, H., additional, MASUDA, Atsushi, additional, NAGAOKA, Takanori, additional, OYAMADA, Masahito, additional, TAKAMATSU, Tetsuro, additional, Furuta, Hirokazu, additional, Hata, Yoshinobu, additional, Yokoyama, Keiichi, additional, Takamatsu, Tetsuro, additional, Itoh, J., additional, Takumi, I., additional, Kawai, K., additional, Serizawa, A., additional, Sanno, N., additional, Teramoto, A., additional, Osamura, R.Y., additional, MATSUTA, Morimasa, additional, MATSUTA, Mayumi, additional, I, Nishiya, additional, TAKAHASHI, Satoru, additional, KAWABE, Kazuki, additional, LIEBER, Michael M., additional, JENKINS, Robert B, additional, SASANO, HIRONOBU, additional, IINO, KAZUMI, additional, SUZUKI, TAKASHI, additional, NAGURA, HIROSHI, additional, Ge, Y-B, additional, Ohmori, J., additional, Tsuyama, S., additional, Yang, D-H, additional, Murata, F., additional, JOHKURA, Kohei, additional, LIANG, Yan, additional, MATSUI, Toshifumi, additional, NAKAZAWA, Ayami, additional, HIGUCHI, Susumu, additional, MATSUSHITA, Yukio, additional, Naritaka, Heiji, additional, Kameya, Toru, additional, Sato, Yuichi, additional, Inoue, Hiroshi, additional, Otani, Mitsuhiro, additional, Kawase, Takeshi, additional, KUROOKA, Yuji, additional, NASU, Kimio, additional, KAMEYAMA, Shuji, additional, MORIYAMA, Nobuo, additional, YANO, Junichi, additional, TSUJIMOTO, Gozo, additional, Matsushita, Tsutomu, additional, Oyamada, Masahito, additional, YAMAMOTO, Hitoshi, additional, MATSUURA, Junko, additional, NOMURA, Takako, additional, SASAKI, Junzo, additional, NAWA, Tokio, additional, KITAZAWA, Riko, additional, KITAZAWA, Sohei, additional, KASIMOTO, Hideyoshi, additional, MAEDA, Sakan, additional, WATANABE, Jun, additional, Mino, Kazuto, additional, KONDO, Kazumasa, additional, KANAMURA, Shinsuke, additional, Ueki, Tetsuo, additional, Takeuchi, Takumi, additional, Nishimatsu, Hiroaki, additional, Kajiwara, Takahiro, additional, Moriyama, Nobuo, additional, Kawabe, Kazuki, additional, Tominaga, Takashi, additional, Kobayashi, Ken-ichiro, additional, Minei, Sadatugu, additional, Okada, Yasuhiro, additional, Yamanaka, Yataro, additional, Ichinose, Taketo, additional, Hachiya, Takahiko, additional, Hirano, Daisaku, additional, Ishida, Hajime, additional, Okada, Kiyoki, additional, HASEGAWA, Hideaki, additional, WATANABE, Keiichi, additional, ITOH, J., additional, HASEGAWA, H., additional, UMEMURA, S., additional, YASUDA, M., additional, TAKEKOSHI, S., additional, OSAMURA, R.Y., additional, WATANABE, K., additional, TAKEDA, Kazuo, additional, HOSHI, Tatsuya, additional, KATO, Katsuaki, additional, OHARA, Shuichi, additional, KONNO, Ryo, additional, ASAKI, Shigeru, additional, TOYOTA, Takayoshi, additional, TATENO, Hiroo, additional, NISHIKAWA, Sumio, additional, SASAKI, Fumie, additional, Ito, Yuko, additional, Matsumoto, Kenji, additional, Daikoku, Eriko, additional, Otsuki, Yoshinori, additional, SANO, Makoto, additional, UMEZAWA, Akihiro, additional, ABE, Hitoshi, additional, FUKUMA, Mariko, additional, SUZUKI, Atsushi, additional, ANDO, Takashi, additional, and HATA, Jun-ichi, additional

Published
1998
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42. Establishment of novel human esophageal cancer cell line in relation to telomere dynamics and telomerase activity.

Author

Kiyozuka, Yasuhiko, Asai, Akira, Yamamoto, Daigo, Senzaki, Hideto, Yoshioka, Sadako, Takahashi, Hakuo, Hioki, Koshiro, Tsubura, Airo, Kiyozuka, Y, Asai, A, Yamamoto, D, Senzaki, H, Yoshioka, S, Takahashi, H, Hioki, K, and Tsubura, A

Subjects
CELL division, COMPARATIVE studies, ESOPHAGEAL tumors, GENES, RESEARCH methodology, MEDICAL cooperation, PROTEINS, RESEARCH, RNA, SQUAMOUS cell carcinoma, TELOMERES, TRANSFERASES, EVALUATION research, CANCER cell culture, NEOPLASTIC cell transformation
Abstract

The telomere and the telomerase in human esophageal cancer are not yet completely understood. The regulatory mechanism of telomerase activity and telomere dynamics has drawn considerable attention. It is generally assumed that when telomerase has been activated, no further telomere shortening should ensue; however, a much more complex pattern of telomere dynamics may exist in telomerase-positive cancer cells. A novel human esophageal cancer cell line (KAN-ES) was established and characterized. Using KAN-ES and its serially passaged subclones up to the 55th generation, we determined the alteration of telomere length (TRF), telomerase activity (TA), telomerase RNA expression (hTR), population doubling time, karyotype, and cytokeratin 14 expression during the process of establishing a cancer cell line. We found that the TRF was maintained between 4.0 and 5.0 kb during the serial passages, despite sustained high TA (assessed by an in vitro TRAP assay). No close relationships were found among TRF, TA, and hTR expression. TA and telomere dynamics were not associated with cellular growth ability and differentiation. However, the number of population doublings showed significant correlations with both the TA and doubling times. In conclusion, these dissociations between telomere dynamics and TA support the existence of additional controls on TRF in cancer cells. KAN-ES and its restored subclones should prove a valuable resource for esophageal cancer research. [ABSTRACT FROM AUTHOR]

Published
2000
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44. An autopsy case of malignant mesothelioma with osseous and cartilaginous differentiation: bone morphogenetic protein-2 in mesothelial cells and its tumor.

Author

Kiyozuka, Y, Miyazaki, H, Yoshizawa, K, Senzaki, H, Yamamoto, D, Inoue, K, Bessho, K, Okubo, Y, Kusumoto, K, and Tsubura, A

Abstract

An autopsy case of biphasic malignant mesothelioma with osseous and cartilaginous differentiation diffusely involving the peritoneal cavity was confirmed by light microscopic histochemistry, immunohistochemistry, and electron microscopy. A reverse transcription-polymerase chain reaction using specific primers for bone morphogenetic protein-2 (BMP-2) revealed weak positive transcript in normal mesothelial cells and up-regulated expression around bone-forming malignant mesothelioma tissue. However, BMP-2 protein expression was detected only in the marginal zone of bone trabeculae and spindle-shaped mesothelioma cells distributed around bone trabeculae in tumor tissue. The distribution of type IV collagen in tumor tissue was in accordance with the BMP-2 expression. Normal mesothelial cells and tumor cells expressed BMP-2 mRNA, but the BMP-2 protein expression was restricted to the bone-forming area in the malignant mesothelioma. [ABSTRACT FROM AUTHOR]

Published
1999

45. Growth inhibitory effects of diallyl disulfide on human breast cancer cell lines.

Author

Nakagawa, H, Tsuta, K, Kiuchi, K, Senzaki, H, Tanaka, K, Hioki, K, and Tsubura, A

Abstract

Diallyl disulfide (DADS) is an oil-soluble organosulfur compound found in garlic. The effect of synthetic DADS on the growth of estrogen receptor (ER)-positive (KPL-1 and MCF-7) and -negative (MDA-MB-231 and MKL-F) human breast cancer cell lines was examined. In an in vitro MTT assay, regardless of ER status, DADS at an IC(50) of 1.8-18.1 microM after 72 h incubation caused inhibition of growth in all four cell lines examined. Growth inhibition was due to apoptosis as seen by the appearance of a sub G1 fraction. In MDA-MB-231 cells, the apoptosis cascade comprised up-regulation of Bax protein (142%), down-regulation of Bcl-X(L) protein (38%) and activation of caspase-3 (438%) compared with controls. In an in vivo assay by orthotopic (right thoracic mammary fat pad) transplantation of KPL-1 cells in female nude mice, intraperitoneal injection of 1 or 2 mg DADS three times a week from the day of tumor cell inoculation until the end of the experiment (after 35 days) caused growth retardation and 43% reductions in primary tumor weight, respectively, compared with DADS-untreated mice without apparent side effects. Cell proliferation as evaluated by proliferating cell nuclear antigen (PCNA)-labeling in transplanted tumor of DADS-untreated mice was 59.6%, and 1 and 2 mg DADS-treated mice was 44.6 and 44.5%, respectively. In MDA-MB-231 cells, DADS antagonized the effect of linoleic acid (LA), a potent breast cancer cell stimulator (at DADS = 1.8 microM and LA > or = 6.5x10(2) microM concentration), and synergized the effect of eicosapentaenoic acid (EPA), a potent breast cancer cell suppressor (at DADS >3 x 10(-3) microM and EPA > 6.3 x 10(-1) microM concentration). Thus, DADS could be a promising anticancer agent for both hormone-dependent and -independent breast cancers, and may harmonize with polyunsaturated fatty acids known as modulators of breast cancer cell growth.

Published
2001
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