Identification of the region in calponin h1 that suppresses proliferation and motility of human fibrosarcoma

钙调蛋白 h1 中抑制人纤维肉瘤增殖和运动的区域的鉴定

• 批准号: 14570117
• 负责人: TAKEOKA Michiko
• 金额: $ 2.24万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570117/
• 关键词: calponin h1; actin; truncate; mutation; cancer; motility; calponin; カルポニンh1; CH domain; Actin binding site; Calponin repeats; C terminal; actin安定性; HT1080細胞; truncates

To investigate the effects of calponin h1(CNh1) on the cell proliferation and motility, human calponin h1 was transfected to human fibrosarcoma, HT1080. CNh1-transfected cells exhibited flattened morphology with organized actin filaments, significant decrease in cell motility. Anchorage-independent growth and tumorigenicity in nude mice were suppressed in CNh1-transfected cells.To identify the region of CNh1 that suppresses proliferation and motility, CNh1 consisted of four domains, CHD, actin binding site (ABS), CNrepeats (CNR) and C terminal, were divided to 12 truncates. As a result, the motility was suppressed by the CNR. Since Repeat 1 (R1) of CNR is reported as a second actin binding site, and has targets of protein kinase C (PKC), actin-depolymerization and podosome formation was investigated. Actin-depolymerization was suppressed by the stimulation with cytochalasin D, and formation of podosome was also suppressed with PDBu (PKC stimulator) in CNh1 transfected cells. Truncates that lack ABS or CNR-R1, and mutant that has mutation of S175A and T184A were transfected. In all three transfectants, transverse actin fiber was separated from calponin h1 and podosome formation was facilitated. Calponin h1 is considered to resist cytochalasin D or PDBu by binding to transverse actin fiber in tumor cells.

为探讨钙调蛋白h1（calponin h1，CNh 1）对人纤维肉瘤HT 1080细胞增殖和运动的影响，将人钙调蛋白h1转染HT 1080细胞，观察CNh 1对HT 1080细胞增殖和运动的影响。CNh 1转染细胞表现出扁平的形态与组织化的肌动蛋白丝，细胞运动性显着下降。CNh 1基因转染细胞后，其生长和致瘤性均受到抑制，CNh 1基因由CHD、actin binding site（ABS）、CN repeats（CNR）和C末端4个结构域组成，分为12个截短片段。结果，CNR抑制了运动。由于CNR的重复序列1（R1）被报道为第二个肌动蛋白结合位点，并且具有蛋白激酶C（PKC）的靶点，因此研究了肌动蛋白解聚和podosome形成。细胞松弛素D的刺激抑制了肌动蛋白解聚，PDBu（PKC刺激剂）也抑制了CNh 1转染细胞中的podosome的形成。转染缺失ABS或CNR-R1的截短体以及具有S175 A和T184 A突变的突变体。在所有三个转染子中，横向肌动蛋白纤维与钙调蛋白h1分离，并促进了podosome的形成。Calponin h1被认为是通过与肿瘤细胞中的横向肌动蛋白纤维结合来抵抗细胞松弛素D或PDBu。

项目成果

Methylation of ASC/TMS1, a proapoptotic gene responsible for activating procaspase-1, in human colorectal cancer.

• DOI: 10.1016/j.canlet.2003.08.027
• 发表时间: 2003-12
• 期刊: Cancer letters
• 影响因子: 9.7
• 作者: Taro Yokoyama;J. Sagara;Xin Guan;J. Masumoto;M. Takeoka;Y. Komiyama;K. Miyata;K. Higuchi;S. Taniguchi

Methylation of ASC/TMS1, a proapoptotic gene responsible for activatinq procaspase-1, in, human colorectal cancer

ASC/TMS1（一种负责激活人类结直肠癌中 procaspase-1 的促凋亡基因）的甲基化

• 发表时间: 2003
• 期刊: Cancer Let 202
• 作者: Yokoyama T;Sagara J;Guan X;Masumoto J;Takeoka M et al.
• 通讯作者: Takeoka M et al.

Calponin h1 suppresses tumor growth of Src-induced transformed 3Y1 cells in association with a decrease in angiogenesis.

• DOI: 10.1111/j.1349-7006.2002.tb01340.x
• 发表时间: 2002-08
• 期刊: Japanese journal of cancer research : Gann
• 作者: Kaneko M;Takeoka M;Oguchi M;Koganehira Y;Murata H;Ehara T;Tozuka M;Saida T;Taniguchi S
• 通讯作者: Taniguchi S

Suppression of peritoneal dissemination through protecting mesothelial cells from retraction by cancer cells

• DOI: 10.1002/ijc.11454
• 发表时间: 2003-11
• 期刊: International Journal of Cancer
• 影响因子: 6.4
• 作者: S. Hashimoto;M. Takeoka;S. Taniguchi

Kaneko M: "Calponin h1 suppresses tumor growth of Src-induced transformed 3Y1 cells in association with a decrease in angiogenesis"Jpn J Cancer Res. 93. 935-943 (2002)

Kaneko M：“Calponin h1 抑制 Src 诱导的转化 3Y1 细胞的肿瘤生长，并与血管生成减少相关”Jpn J Cancer Res。