Extracellular Release of Annexin II From Pancreatic Cancer Cells and Resistance to Anticancer Drug–Induced Apoptosis by Supplementation of Recombinant Annexin II
Extracellular Release of Annexin II From Pancreatic Cancer Cells and Resistance to Anticancer Drug–Induced Apoptosis by Supplementation of Recombinant Annexin II
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DOI：
10.1097/mpa.0b013e31824f356f
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发表时间：
2012-11
期刊：
影响因子：
2.9
通讯作者：
Tetsuo Sato;K. Kita;S. Sugaya;Toshikazu Suzuki;N. Suzuki
中科院分区：
文献类型：
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作者：
Tetsuo Sato;K. Kita;S. Sugaya;Toshikazu Suzuki;N. Suzuki
目的细胞外微环境在肿瘤的发生、发展和耐药过程中起重要作用。胰腺癌对几乎所有的化疗药物都有耐药性。在这项研究中，我们确定了膜联蛋白II在培养基中从胰腺癌细胞释放到细胞外环境的蛋白质。方法收集各种癌细胞培养5小时的培养基。通过分子量分析和免疫印迹检测培养基中的蛋白质。用结晶紫法和集落存活法检测重组膜联蛋白II（rANX II）预孵育前后细胞对抗癌药物的敏感性。通过免疫印迹分析凋亡相关分子。结果培养基中添加重组ANX II可使MiaPaCa-2和AsPC-1细胞对顺铂、5-氟尿嘧啶和吉西他滨等抗癌药物产生耐药性。在MiaPaCa-2细胞中，rANX II补充导致与Bcl-2/Bax比率增加相关的caspase-3活化的抑制。通过补充rANX II抑制顺铂诱导的细胞死亡被磷脂酰肌醇3-激酶和丝裂原活化蛋白激酶激酶信号通路的抑制剂取消。结论本研究首次证明培养基中添加rANX II可增加胰腺癌细胞对抗癌药物的耐药性。重组ANX II通过拮抗顺铂诱导的细胞凋亡发挥细胞死亡抑制功能。
Objectives Extracellular microenvironment plays crucial roles in the development of cancers and chemoresistance. Pancreatic carcinoma is resistant to almost all chemotherapeutic agents. In this study, we identified annexin II in the medium from pancreatic cancer cells as a protein released into the extracellular environment. Methods Medium from 5-hour cultures of various cancer cells was collected. Proteins in the medium were detected by molecular mass analysis and immunoblotting. Anticancer drug sensitivity of cells preincubated with or without recombinant annexin II (rANX II) was measured using crystal violet assay and colony survival assay. Apoptosis-related molecules were analyzed by immunoblotting. Results Recombinant ANX II supplementation in the medium confers resistance to anticancer drugs, including cisplatin, 5-fluorouracil, and gemcitabine, in MiaPaCa-2 and AsPC-1 cells. In MiaPaCa-2 cells, rANX II supplementation resulted in suppression of caspase-3 activation associated with increased Bcl-2/Bax ratios. Suppression of cisplatin-induced cell death by rANX II supplementation was canceled by inhibitors of phosphatidylinositol 3-kinase and mitogen-activated protein kinase kinase signal pathways. Conclusions The current study is the first report to demonstrate that supplementation of rANX II in the medium increased resistance to anticancer drugs in pancreatic cancer cells. Recombinant ANX II exerts cell death–suppressive function by antagonizing cisplatin-induced apoptosis.