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Carcinogenesis, Vol. 20, No. 3, 465-469, March 1999
© 1999 Oxford University Press

Overexpression of midkine in lung tumors induced by N-nitrosobis(2-hydroxypropyl)amine in rats and its increase with progression

Hiroyuki Sakitani1,2, Masahiro Tsutsumi1,5, Kenji Kadomatsu3, Shinya Ikematsu4, Makoto Takahama1, Katsumichi Iki1, Toshifumi Tsujiuchi1, Takashi Muramatsu3, Sdatoshi Sakuma4, Toshisuke Sakaki2 and Yoichi Konishi1

1 Department of Oncological Pathology, Cancer Center,
2 Department of Neurosurgery, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521,
3 Department of Biochemistry, Nagoya University School of Medicine, 65 Tsurumai-cho, Syowa-ku, Nagoya, Aichi 466-8550 and
4 Meiji Cell Technology Center, 540 Naruda Odawara 250-086, Japan

The expression of midkine (MK) in lung tumors induced by N-nitrosobis(2-hydroxypropyl)amine (BHP) in rats was examined. The animals were administered 2000 p.p.m. of BHP in their drinking water for 12 weeks, then maintained without further treatment until being killed 20–28 weeks after the beginning of the experiment. MK mRNA expression of adenocarcinomas and squamous cell carcinomas assessed by means of the reverse transcriptase–polymerase chain reaction and northern blot analysis was significantly higher than in rat embryonic tissues (positive controls) and contrasted strongly with the lack in normal lungs. MK protein was detected immunohistochemically in 58.3% of alveolar hyperplasias, 92.3% of adenomas and 100% of adenocarcinomas and squamous cell carcinomas. The extent of staining significantly increased along with malignant progression in adenomatous (pre-)neoplastic lesions and tended to become more pronounced with malignant progression in squamous lesions. The results suggest that MK may play some essential roles in the development and progression of lung tumors induced by BHP in rats.

Abbreviations: BHP, N-nitrosobis(2-hydroxypropyl)amine; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; MK, midkine; RT–PCR, reverse transcriptase–polymerase chain reaction; SDS, sodium dodecyl sulfate; SSC, standard saline citrate.

5 To whom correspondence should be addressed Email: hyro-sa{at}nmu-gw.cc.naramed-u.ac.jp


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