利用JC類病毒殼體攜帶anti-SV40 LT shRNA抑制人類腦瘤細胞生長之研究

Growth inhibition of glioma cells (SVGA) by anti-SV40 LT shRNA delivered by the JC virus-like particle

黃育甫(Yufu Huang)

指導教授 ： 張德卿

國立中正大學/理學院/生命科學系分子生物研究所/碩士(2013年)

摘要

人類多瘤性JC病毒，能夠感染人類少突膠質細胞並造成致死性的多病灶腦白質病變(progressive multifocal leucoencephalopathy , PML)疾病，JC病毒殼體，可能可作為一基因輸送載體，以針對神經性疾病達到治療之目的。先前的研究發現JC病毒之VP1結構蛋白可藉由E.coli系統表達，並可自行組裝為病毒類似殼體(Virus like particle, VLP)，實驗室過去也證實，利用JC類病毒殼體去攜帶Oligodeoxynucleotide (ODN)進入人類的星形細胞瘤、神經母細胞瘤和膠質母細胞瘤細胞內，皆有高效率之表達， 結果顯示JC VLP對於神經細胞的確是一個很有潛力的基因輸送載體。在我們這次的研究中設計了一個會表達抑制Simian virus 40 large tumor antigen (SV40 LT)致癌蛋白的short hairpin RNA (shRNA)之質體，並確定其可抑制SVGA細胞之SV40 LT表達，接著便將此質體包裹於JC VLP中(Shs1LT-VLP and Shs4LT-VLP)並運送至細胞，結果顯示其的確具有抑制SV40 LT致癌蛋白表達之趨勢。而利用類似的方法及技術可望於未來為人類腦瘤治療方式帶來新的方向。

關鍵字

多瘤性病毒

並列摘要

Human JC virus (JCV) is able to infect human oligodendrocytes and cause a fatal disease, progressive multifocal leukoencephalopathy (PML). It is possible to use the JCV capsid as a gene-transduce vector for therapeutic purposes in neurological disorders. In our previous studies it has also been found that JCV VP1 self-assembles into a virus-like particle (VLP) structure when expressed in E. coli. Previously, it has been demonstrated that the JCV VLP was able to deliver ODN into human astrocytoma, neuroblastoma, and glioblastoma cells with high efficiency. The results indicate that the JCV VLP generated in E coli could potentially be used as a gene dilivery vector. In this study, we constructed a plasmid expressing anti-SV40 LT short hairpin RNA (shRNA). In vitro, the shRNA plasmid could inhibit SV40 LT expression in SVGA cells. The anti-LT shRNA plasmid can be packaged and delivered by the JC VLP to inhibit the SV40 LT expression human glioma cells as demonstrated in this study. Therefore it's possible that human brain tumor oncogenes could be inhibited by the similar approaches for brain tumor therapy in the future.

並列關鍵字

polyomavirus

參考文獻

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