探討第二型拓撲異構酶抑制劑增強桿狀病毒外源基因表達之機制以降低化療時藥物所產生的副作用之可行性

1983年Smith等人對於核多角體病毒開發成真核細胞表現載體 (eukaryotic expressing vector) 進行研究，並成功生產具有活性的人類β-干擾素 (human β-interferon) ，此後，桿狀病毒表現系統被廣泛探討與研究。雖然如此，在早期，對於桿狀病毒表現系統的研究多侷限於無脊椎動物的模型中，直到1995年 Christian Hofmann 的研究團隊發現桿狀病毒可把外來基因送入人類的肝癌細胞，也能於細胞內表現報導基因亦即表示桿狀病毒可以有效地利用哺乳動物細胞啟動子。因此，科學家開始利用桿狀病毒表現系統對哺乳動物細胞進行臨床前的研究，其中，在2010 Haiyan Guo的研究團隊利用桿狀病毒攜帶 p53 基因，並利用 NaBt (一種組蛋白去乙醯化抑制劑，可增強桿狀病毒外源基因表達)使 p53 大量表達來殺死癌細胞。另外，在先前研究顯示第二型拓撲異構酶的抑制劑也能增強桿狀病毒的外源基因表達。由於 HDAC 在亞型上類別較多，且在不同類型的細胞中抑制不同的 HDAC 會造成諸多影響，在藥物作用上需注意較多，但拓撲異構酶第二型在體細胞內較為單純，因此本實驗希望能透過第二型拓撲異構酶抑制劑－多柔比星 (Doxorubicin) 針對受攜帶 p53 外源基因的桿狀病毒所感染的骨肉瘤細胞進行研究，了解其增強基因表達的機制並探討其降低化療藥物於化療時所產生的副作用的可行性。

關鍵字

桿狀病毒；第二型拓樸異構酶抑制劑；多柔比星

並列摘要

In 1983, Smith and his colleague developed the overexpression of human IFN-β in insect cells with recombinant baculovirus. This is one of the reasons baculovirus expression system has become one of the most widely used systems for production of recombinant proteins. Nonetheless, in the early baculovirus expression system for the study was restricted to invertebrate model. Until 1995, Christian Hofmann's research team found that baculovirus can be foreign genes into human liver cancer cells, but also in the cell performance of the gene that is baculovirus can be the quite effective use of mammalian cell promoter. As a result, scientists have begun to use baculovirus expression systems to conduct preclinical studies of mammalian cells, in which the study team at 2010 Haiyan Guo used baculovirus to carry the p53 gene and added NaBt (a histone deacetylation inhibition agent, can enhance the expression of baculovirus exogenous gene) to enable to express a large number of p53 in cancer cells. In addition, previous studies have shown that inhibitors of type II topoisomerase also enhance the expression of exogenous genes of baculoviruses. Because HDAC in the subtypes on the more categories, and in different types of cells to inhibit the different HDAC will cause many effects in the role of drugs should pay more attention to, but the type II topoisomerase in vivo cells more simple. Therefore, we expects to study the osteosarcoma cells infected by baculoviruses carrying p53 exogenous gene through the type II topoisomerase inhibitor, Doxorubicin, enhance the mechanism of gene expression and explore the feasibility of reducing the side effects of chemotherapy drugs in chemotherapy.

並列關鍵字

Baculovirus ； Type II topoisomerase inhibitor ； Doxorubicin

參考文獻

(2016). Topoisomerase inhibitors. 72-85.

Aggarwal, S.K., Goel, D., Gupta, C.R., Ghosh, S., and Ojha, H. (2002). The use of pedicled appendix graft for substitution of urethra in recurrent urethral stricture. J Pediatr Surg 37, 246-250.

Airenne, K.J., Hu, Y.C., Kost, T.A., Smith, R.H., Kotin, R.M., Ono, C., Matsuura, Y., Wang, S., and Yla-Herttuala, S. (2013). Baculovirus: an insect-derived vector for diverse gene transfer applications. Mol Ther 21, 739-749.

Allfrey, V.G., Faulkner, R., and Mirsky, A.E. (1964). Acetylation and Methylation of Histones and Their Possible Role in the Regulation of Rna Synthesis. Proc Natl Acad Sci U S A 51, 786-794.

Brownell, J.E., Zhou, J., Ranalli, T., Kobayashi, R., Edmondson, D.G., Roth, S.Y., and Allis, C.D. (1996). Tetrahymena histone acetyltransferase A: a homolog to yeast Gcn5p linking histone acetylation to gene activation. Cell 84, 843-851.

國際替代計量

探討第二型拓撲異構酶抑制劑增強桿狀病毒外源基因表達之機制以降低化療時藥物所產生的副作用之可行性

全文下載

主題瀏覽