CHM-1誘導 SIRT2 表現抑制人類乳癌細胞之增生

CHM-1 inhibit human breast cancer cells by inducing SIRT2 gene expression

劉晉瑋(Chin-Wei Liu)

指導教授 ： 洪志勳

義守大學/理工學院/化學工程學系暨生物技術與化學工程研究所/博士(2021年)

摘要

乳癌為全球女性共通的惡性腫瘤，乳癌早期篩檢、治療及相關藥物的研究是相當重要的。目前新型合成藥物 CHM-1 (2-(2-fluorophenyl)-6,7-methylenedioxoquinolin-4-one)，可與微管蛋白相互作用，顯著抑制微管蛋白聚合，並破壞微管組織，將用來探討對於人類乳腺癌的抑制效果。本實驗以不同濃度之CHM-1處理 MDA-MB-231、MDA-MB-453 和 MCF-7 細胞，發現皆有抑制效果，其中對MCF-7 細胞最顯著，而實驗也發現 CHM-1 對正常乳腺 ( HBL-100 ) 細胞的毒性較小。通過二維電泳和 MALDI-TOF 質譜分析CHM-1 處理過的動物腫瘤樣品與對照組的蛋白質，發現SIRT2、vimentin、 和 ACTB 蛋白有顯著上升。再以 CHM-1處理 MCF-7 細胞後，確實可增加 SIRT2 表現量。SIRT2 蛋白是一種 NAD 依賴性微管蛋白去乙醯酶，可將 -tubulin 蛋白去乙醯化。此外，我們還合成前趨藥CHM-1-P，以解決抗腫瘤劑 CHM-1 的低水溶性和低生物利用度的問題。SCID小鼠口服CHM-1-P 28天後，會抑制 MCF-7 腫瘤生長 (20 mg/kg CHM-1-P 處理組體積為 177.93 mm3，相對於對照組 P <0.001；對照組在第 28 天時體積為 2113.28 mm3)。對SCID小鼠每週進行兩次靜脈注射 CHM-1-P ( 20 mg/kg )，經過 28 天後發現可導致腫瘤體積減少 95 % ( P <0.001)。上述 CHM-1 及其前驅藥 CHM-1-P 的優異抗腫瘤活性從結果顯示，CHM-1與 CHM-1-P 值得進一步發展成為治療乳癌的臨床試驗候選者。

關鍵字

抗有絲分裂劑 ； 前趨藥

並列摘要

Breast cancer is a malignant tumor common to women all over the world. Early breast cancer screening, treatment and research on related drugs are very important. The current new synthetic drug CHM-1 (2-(2-fluorophenyl)-6,7-methylenedioxoquinolin-4-one), which can interact with tubulin, significantly inhibit tubulin polymerization and destroy microtubule tissue, will be used to explore the inhibitory effect on human breast cancer. In this experiment, different concentrations of CHM-1 were used to treat MDA-MB-231, MDA-MB-453, and MCF-7 cells, and all of them were found to have inhibitory effects. Among them, MCF-7 cells were the most significant, and the experiment also found that CHM-1 are less toxicity to normal breast (HBL-100) cells. By two-dimensional electrophoresis and MALDI-TOF mass spectrometry analysis of CHM-1 treated animal tumor samples and control group proteins, it was found that SIRT2, vimentin, and ACTB proteins were significant upregulation. After CHM-1 treatment of MCF-7 cells, it can indeed increase the expression of SIRT2 from Western blot analysis. SIRT2 protein is a NAD-dependent tubulin deacetylase that can deacetylate -tubulin protein. In addition, we also synthesized the prodrug CHM-1-P to solve the problem of low water solubility and low bioavailability of the antitumor agent CHM-1. After 28 days of oral administration of CHM-1-P to SCID mice, it will inhibit the growth of MCF-7 tumors (the volume of the 20 mg/kg CHM-1-P treatment group is 177.93 mm3, which is P <0.001 relative to the control group; the control group has the volume on the 28th day 2113.28 mm3). SCID mice were given intravenous injection of CHM-1-P (20 mg/kg) twice a week. After 28 days, it was found that the tumor volume decreased by 95%. (P<0.001). The excellent anti-tumor activity of the above-mentioned CHM-1 and its prodrug CHM-1-P. The results show that CHM-1 and CHM-1-P are worthy of further development as candidates for clinical trials for the treatment of breast cancer.

並列關鍵字

CHM-1 ； SIRT1 ； SIRT2 ； Antimitotic agent ； Prodrug

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