環狀 RGD 胜?誘導乳癌細胞株 MCF-7 細胞凋亡的磷酸化蛋白質探討

Abstract

在許多配位分子中發現一種 integrin 的辨識區域，稱為 RGD motif。因此包含 RGD motif 的胜?可以和 integrin 受體結合，並進而引起細胞訊息的傳遞和影響許多疾病的過程。人工合成含有 RGD motif 的胜?已經廣泛被使用作為 ingegrin-ligand 相互作用的抑制劑，以進一步研究細胞的生長、死亡、附著、遷移和侵略。因此，我們設計了一個含有環狀 RGD 的胜? (Tpa-RGDWPC)，有著更堅固的分子骨幹可以和受體緊密結合。根據之前的文獻報導指出，cRGD 可以誘導 MCF-7 乳癌細胞株的死亡。我們對於以 cRGD 處理 MCF-7 細胞的分子機制很有興趣。此外，因為後轉譯修飾的蛋白質磷酸化在細胞傳遞訊息中扮演極重要的角色，因此，我們使用蛋白質體學結合 Pro-Q Diamond 磷酸化蛋白染劑技術來作以 cRGD 處理過後的 MCF-7 細胞的整體性細胞蛋白觀察。在本次實驗中，我們找到一些磷酸化蛋白在 cRGD 誘導 MCF-7 細胞死亡中扮演重要角色，可做為一些治療乳癌的治療開發基礎。

The RGD (arginine-glycine-aspartate) motif is an integrin-recognition motif found in many ligands, so that RGD-containing peptides can interact with the integrin receptor to induce cell-signaling processes and influence many different diseases. Synthetic peptides containing the RGD motif have been used extensively as inhibitors of integrin-ligand interactions in studies of cell growth, death, adhesion, migration, and invasion. Therefore, we designed a cyclic-RGD peptide (Tpa-RGDWPC, cRGD) with rigid skeleton to closely bind with its receptor. According to previous reports that cRGD can cause cell death in MCF-7 breast cancer cell line. We are interested in what molecular mechanism underlies the cRGD exerts on MCF-7 cells. Depending on protein phosphorylation, it is a major post-translational modification connected to the signaling pathways in the cell. We used the proteomics with Pro-Q Diamond phosphoprotein dye technology to survey the global changes in phosphoproteins after cRGD treatment in MCF-7 cells. In this report, we find some phosphoproteins play important roles in cRGD treated MCF-7 cell death and provide a valuable in-depth insight into its use in breast cancer therapy.