| 摘要: | Mitogen-Activated protein kinase (MAPK) 活化後，進行一連串複雜的細胞反應，如細胞增生、分化及細胞凋亡等。 Ribosomal S6 kinase (RSK)，在MAPK 路徑中扮演重要角色，經由磷酸化調控許多細胞質與細胞核蛋白質的功能，亦會調節一些轉錄因子，使得細胞增生、分化或存活等作用，並有文獻指出 RSK 在乳癌和前列腺癌中亦會大量表現。文獻報告指出 BI-D1870為RSK 抑制劑之一， 顯示對 RSK 有專一性的細胞傳遞之抑制活性，且為 ATP 競爭型抑制劑。本論文探討 BI-D1870 對於口腔癌細胞的生長，細胞週期調控及凋亡因子的影響，利用 MTT assay 和 Cell counting 測試 BI-D1870對於口腔癌細胞 (SCC2095, SCC4, SCC9, Ca922, and HSC3) 及正常人類口腔上皮細胞 (Normal human keratinocytes) 的生長抑制。並利用流式細胞儀偵測口腔癌細胞的細胞週期、細胞凋亡、以及活性氧基群的表現，同時也利用 DAPI staining 偵測細胞核質濃縮現象。結果顯示 BI-D1870 會抑制口腔癌細胞的生長，對於口腔癌細胞，SCC4 cells 最具毒殺效果，IC50為1.7 ?嵱，並造成細胞週期停滯在 G2/M 期，進而造成細胞凋亡。同時也會促使粒線體凋亡路徑及細胞週期相關蛋白的活化，除此之外，BI-D1870 亦會抑制 RSK 之受質與細胞增生相關的蛋白表現量。
Mitogen-activated protein kinase (MAPK) executes complex cellular programs, like proliferation, differentiation and apoptosis. Ribosomal S6 kinase (RSK), a key player in MAPK pathway, modulates the activities of several cytoplasmic and nuclear proteins via phosphorylation. It also regulated some transcription factors, lead to the cell proliferation, differentiation and survival. And there are some papers to show that RSK was overexpression in breast cancer and prostate cancer. It has been reported recently, that BI-D1870, one of RSK inhibitors, showed the specific cell-permeant inhibitory activity of RSK, which acts as an ATP competitive inhibitor. In this study, we explore the effects of BI-D1870 for cell growth, cell cycle and apoptosis on oral cancer cells. We used MTT assay and Cell counting test to detect cell survival in oral cancer cells (SCC2095, SCC4, SCC9, Ca922, and HSC3) and normal human keratinocytes. Then flow cytometry was used to detect cell cycle and apoptosis, and the activity of reactive oxygen species in oral cancer cells, DAPI staining was also used to detect nuclear concentration. Results show that BI-D1870 inhibits oral cancer cells growth inducing cell cycle arrest in the G2/M phase, and causes apoptosis. In addition, BI-D1870 also inhibits RSK and cell proliferation-related protein expression. |
