effect of the inhibitor to membrane-bound enzyme on the growth of invasion of cancer cells

膜结合酶抑制剂对癌细胞侵袭生长的影响

• 批准号: 06671642
• 负责人: GOTO Setsuko
• 金额: $ 1.28万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06671642/
• 关键词: Ubenimex (bestatin); cell-growth suppression; aminopeptidase N; Monoclonal antibody; Immunoblotting; MTT assay; bestatin uptake; flow cytometry; DNAhistogram; アミノペプチダーゼN; MMT assay

We previously found that ubenimex (bestatin), an aminopeptidase inhibitor, had a growth-suppressive effect on cancer cells. To clarify the mechanism of the growth-suppressive effect, we investigated the expression of aminopeptidase N (AP-N/CD13) on choriocarcinoma cells and other human tumor cells.1) Two choriocarcinoma cell lines, NaUCC-4 and BeWo, had higher AP-N activity than other cell lines, as did the human myeloid leukemia cell line, IIL-60.2) These choriocarcinoma and leukemia cell lines with aboundunt AP-N activity showed much higher sensitivity to bestatin than the other cell lines. 3) By immunoblotting and immunocytochemical staining, AP-N was detected as an approximately 165-kDa protein and localized on the cell membrane in choriocarcinoma cells. 4) We examined the effects of three anti-CD13 monoclonal antibodies (WM15, MCS2, and MY7) on the growth of NaUCC-4 cells. Of the three MABs, only WM15, which is able to inhibit AP-N activity, suppressed cell growth in a dose-dependent manner.These results indicate that AP-N inhibitors show a growth-suppressive effect, presumably through inhibition of the enzyme activity of AP-N on carcinoma cells.5) we examined bestatin uptake using 3H-bestatin in choriocarcinoma cells and other cells. A higher upake of 3H-bestatin was resulted in NaUCC-4 cells, which showed the highest sensitivity bestatin. 6) In DNA histogram, NaUCC-4 cells treated with bestatin were not blocked at any cell-cycle phases as studied by flow fluorocytometry.

我们先前发现氨基肽酶抑制剂乌苯美司(BESTATIN)对癌细胞有生长抑制作用。1)两株绒癌细胞株NaUCC-4和BeWo细胞株AP-N活性高于其他细胞株，人髓系白血病细胞株IIL-60.2细胞株AP-N活性明显高于其他细胞株。3)免疫印迹和免疫细胞化学染色检测到绒毛膜癌细胞AP-N蛋白的大小约为165 kDa，定位于细胞膜。4)检测了三种抗CD13单抗(WM15、MCS2和MY7)对NaUCC-4细胞生长的影响。在三种单抗中，只有能够抑制AP-N活性的WM15以剂量依赖的方式抑制细胞生长。这些结果表明，AP-N抑制剂显示出生长抑制作用，可能是通过抑制AP-N对癌细胞的酶活性。5)我们用~H-Bestatin检测了绒毛膜癌细胞和其他细胞对Bestatin的摄取。NaUCC-4细胞的~3H-Bestatin水平较高，对BEST的敏感性最高。6)在DNA直方图中，Bestatin处理的NaUCC-4细胞没有被流式细胞仪检测到的任何细胞周期时相被阻断。

项目成果

Kazuhiko Ino: "Expression of Aminopeptidase N on Human Choriocarcinoma Cells and Cell Growth Supression by the Inhibition of Aminopeptidase N Activity." Jpn.J.Cancer Res.85. 927-933 (1994)

Kazuhiko Ino：“氨肽酶 N 在人绒毛膜癌细胞上的表达以及通过抑制氨肽酶 N 活性来抑制细胞生长。”

Kazuhiko Ino: "Experssion of Aminopeptidase N on Human Choriocarcinoma Cells and Cell Growth Supression by the Inhibition of Aminopeptidase N Activity" Jpn.J.Cancer Res.85. 927-933 (1994)

Kazuhiko Ino：“氨基肽酶 N 在人绒毛膜癌细胞上的表达以及通过抑制氨基肽酶 N 活性来抑制细胞生长”Jpn.J.Cancer Res.85。

Kazuhiko Ino, Setsuko Goto, Tomomitu Okamoto, Seiji Nomura, Akihiro Nawa, Ken-ichi Isobe, Shigehiko Mizutani, Yutaka Tomoda.: "Expression of Aminopeptidase N on Human Choriocarcinoma Cells and Cell Growth Suppression by the Inhibition of Aminopeptidase N

Kazuhiko Ino、Setsuko Goto、Tomomitu Okamoto、Seiji Nomura、Akihiro Nawa、Ken-ichi Isobe、Shigehiko Mizutani、Yutaka Tomoda.：“氨肽酶 N 在人绒毛膜癌细胞上的表达以及通过抑制氨肽酶 N 抑制细胞生长