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題 名 | Targeting Transforming Growth Factor-Betal Using RNA Interference Attenuates Melanoma Cell Growth in an Ex-vivo Animal Model=針對轉型生長因子β1進行核糖核酸干擾抑制作用能縮減黑色素瘤細胞在體內生長 |
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作 者 | 戴國峰; 曾斯偉; 朱小芬; 劉安邦; 王健興; | 書刊名 | 慈濟醫學 |
卷 期 | 19:1 2007.03[民96.03] |
頁 次 | 頁5-12 |
分類號 | 415.691 |
關鍵詞 | 轉型生長因子; 核糖核酸干擾抑制; 黑色素瘤細胞; RNA interference; TGF-β1; Melanoma; |
語 文 | 英文(English) |
英文摘要 | Objective: The transforming growth factor-beta(TGF-β) is the key molecule implicated in impaired immune function in human patients with malignant melanoma. We examined the effects of TGF-β1protein knockdown using RNA interference on the growth of melanoma in C57BL/6 mice induced using the B16F0 cell. Materials and Methods: The TFG-β1 hairpin oligonucleotide was lconed into retroviral vector pSM2. The resulting plasmid (TFG-β1-RNAi/pSM2) was stably introduced into the murine melanoma cell line, B16F0, and designated as B16F0/ TFG-β1-RNAi cells. The vector plasmid was transfected into B16F0 cells and designed as B16F0/vector-control cells which served as the control. Five million B16F0 cells, B16F0/vector-control cells and B16F0/ TFG-β1-RNAi cells were injected subcutaneously into the right flanks of adult female syngeneic mice C57BL/6, respectively. The growth rate of the parental cells and genetically modified murine melanoma cells were compared. Results: TFG-β1 expression was reduced in B16F0/ TFG-β1-RNAi cells compared with B16F0 cells and B16F0/vector-control cells. The proliferation rate of the B16F0/ TFG-β1-RNAi cells was similar to that of the B16F0 cells and B16F0/vector-control cells in vitro. The tumor sizes were 495.32±77.25m□, 516.65±73.71m□ and 326.72±97.34m□ on the 14th day after the mice received B16F0 cells. B16F0/vector-control cells and B16F0/ TFG-β1-RNAi cells, respectively. The pvalue was less than 0.05 using one-way ANOVA. TFG-β1knockdown in B16F0 cells enhanced the infiltration of CD□ and CD□ T cells in the tumor regions. The blood vessel density of the tumors was markedly reduced in the B16F0/ TFG-β1-RNAi tumors. Conclusions: We found that TFG-β1 protein expression was significantly reduced from B16F0 cells after TFG-β1 hairpin oligonucleotide transduction. Silencing of TFG-β1 expression in B16F0 cells using RNA interference technology inhibited the growth of this tumor after being transplanted to C57BL/6 mice. Overall, our results have important implications for the clinical use of RNA interference targeting TFG-β1 as cancer gene therapy. |
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