Analysis of mechanisms of apoptosis resistance in pancreatic cancer cells

胰腺癌细胞抗凋亡机制分析

• 批准号: 18590745
• 负责人: HIGUCHI Hajime
• 金额: $ 2.51万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590745/
• 关键词: side population; cancer stem cell; invation; epithelial to mesenchymal transition

This study evaluated the mechanisms of biological malignant potential, i.e., apoptosis resistance, growth potential, activity of invasion and metastasis, in pancreatic cancer cells. We first focused on epidermal growth factor receptor expression and mutasion in pancreatic cancer cell lines such as Panc-1, MIA PaCa-2, Capan-1, Capan-2 and BxPC-3. We did not find positive correction between apoptosis resistance and EGFR expression levels. In addition, we did not find specific EGFR mutation that correlates apoptosis resistance in all the pancreatic cancer cell lines used in this study. Therefore, we next focused on heterogeneity of the cells within each cell line. We identified both side population(SP), an apoptosis resistant fraction, and main population(MP), a relatively sensitive population to chemotherapy-induced apoptosis in some pancreatic cancer cell line. By comparative analysis of SP cells and MP cells, SP cells were resistant to a variety of apoptosis-including stimuli, i.e., tu … More mor necrosis factor-related ligand(TRAIL), 4-fluorouracil(5-FU) and cisplatin(CDDP). In addition, SP cells displayed greater in vitro colony forming activity and in vivo tumor forming capacity, suggesting that SP cells have so-called cancer stem cell-like properties. In an isolated SP cell culture, the cells rapidly expressed and upregulated E-cadherin, an epithelial phenotypic marker, and the cells grew up forming tightly contained cell cluster, which is a representative epithelial phenotypic appearance. When the SP cells were incubated in the presence of TGF-beta, the SP cells changed their shape into mesenchymal-like appearance including spindle shaped assembly. This alteration was concordant with significant reduction of E-cadherin protein expression level. TGF-beta induced EMT-associated gene alteration such as reduction of E-cadherin mRNA and induction of Snail mRNA and matrixmetalloproteinase (MMP)-2 mRNA. SP cells appears to respond TGF-b-signaling more, as TGF activin responsive element (TARE)-based luciferase reporter assay revealed superior response to TGF-beta treatment in SP cells as compared to MP cells. Finally, SP cells exerted notable matrigel invasion activity in response to TGF-beta, while MP cells did not respond to TGF-beta-mediated invasion. Because SP fraction is known as cancer stem cell-enriched fraction, we conclude that SP cells play a major role in apoptosis resistance in pancreatic cancer cells. SP cells also play a important roles in pancreatic cancer invasion and metastasis. Thus, SP cells appears to be a target to prevent cancer development. Less

本研究探讨了胰腺癌细胞生物学恶性潜能的机制，即凋亡抵抗、生长潜能、侵袭和转移活性。我们首先研究了表皮生长因子受体在胰腺癌细胞系PANC-1、MIA PACA-2、CAPAN-1、CAPAN-2和BxPC-3中的表达和突变。我们没有发现凋亡抵抗与EGFR表达水平之间的正相关。此外，我们没有在本研究中使用的所有胰腺癌细胞系中发现与凋亡抵抗相关的特定的EGFR突变。因此，我们接下来关注每个细胞系内细胞的异质性。我们在一些胰腺癌细胞系中发现了对化疗诱导的细胞凋亡相对敏感的侧群(SP)和主群(MP)。通过对SP细胞和MP细胞的比较分析，发现SP细胞对包括TU-…在内的多种细胞凋亡均有抵抗作用更多的是肿瘤坏死因子相关配体(TRAIL)、4-氟尿嘧啶(5-FU)和顺铂(CDDP)。此外，SP细胞在体外表现出较强的集落形成活性和体内成瘤能力，提示SP细胞具有肿瘤干细胞样特性。在分离培养的SP细胞中，细胞迅速表达并上调上皮表型标志物E-钙粘蛋白，细胞生长形成紧密包裹的细胞团，这是典型的上皮表型特征。当SP细胞与转化生长因子-β共同孵育时，SP细胞的形态转变为间充质样外观，包括梭形集合体。这一变化与E-钙粘蛋白蛋白表达水平的显著降低相一致。转化生长因子-β诱导内皮细胞转化相关基因改变，如E-钙粘蛋白基因表达降低，Snail基因和基质金属蛋白酶-2基因表达上调。SP细胞似乎对转化生长因子-β信号的反应更多，因为基于转化生长因子激活素反应元件(TARE)的荧光素酶报告分析显示，与MP细胞相比，SP细胞对转化生长因子-β处理的反应更好。最后，SP细胞对转化生长因子-β具有显著的基质侵袭活性，而MP细胞对转化生长因子-β介导的侵袭没有反应。由于SP组分被称为肿瘤干细胞富集组分，因此我们得出结论，SP细胞在胰腺癌细胞的凋亡抵抗中发挥着重要作用。SP细胞在胰腺癌的侵袭和转移中也起着重要作用。因此，SP细胞似乎是预防癌症发展的靶点。较少

项目成果

MCL-1 depletion sensitizes the apoptosis of gastrointestinal cancer cells mediated by anti-cancer drugs

MCL-1耗竭使抗癌药物介导的胃肠道癌细胞凋亡敏感

• 发表时间: 2007
• 作者: Hideko Iizuka;Hajime Higuchi;Hidetoshi Sumimoto;Ayano Kabashima;Naoko Kuriyama;Gen Sakai;Motoko Izumiya;Yoshiyuki Yamagishi;Hiromasa Takaishi;Yutaka Kawakami;Hiroyuki Mizuguchi;Toshifumi Hibi
• 通讯作者: Toshifumi Hibi

Transforming growth factor(TGF)-bate induces pancreatic cancer cell invasion and epithelial to mesenchymal(EMT)in side-population(SP)cells but not in non SP cells

转化生长因子 (TGF)-bate 在侧群 (SP) 细胞中诱导胰腺癌细胞侵袭和上皮间质 (EMT)，但在非 SP 细胞中则不然

• 发表时间: 2007
• 作者: Kabashima A;Higuchi H;Matsuzaki Y;Hasegawa G;Kuriyama N;Takaishi H;Izumiya M;Iizuka H;Sakai G;and Hibi T.
• 通讯作者: and Hibi T.

Transforming growth factor(TGF)-bate induces pancreatic cancer cell invasion and epithelial to mesenchymal (EMT)in side-population(SP)cells but not in non SP cells.

转化生长因子 (TGF)-bate 在侧群 (SP) 细胞中诱导胰腺癌细胞侵袭和上皮间质 (EMT)，但在非 SP 细胞中则不然。

• 发表时间: 2007
• 作者: Kabashima;A.;Higuchi;H.;Matsuzaki;Y.;Hasegawa;G.;Kuriyama;N.;Takaishi;H.;Izumiya;M.;Iizuka;H.;Sakai;G.;Hibi;T
• 通讯作者: T

Mcl-l depletion sensitizes the apoptosis of gastrointestinal cancer cells mediated by anti-cancer drugs.

Mcl-1 耗竭使抗癌药物介导的胃肠道癌细胞凋亡变得敏感。

• 发表时间: 2007
• 作者: Iizuka;H.;Higuchi;H.;Sumimoto;H.;Kabashima;A.;Kuriyama;N.;Sakai;G.;Izumiya M.;Ymagishi;Y.;Takaishi;H.;Kawakami Y;Mizoguchi;H.;Hibi T
• 通讯作者: Hibi T