CS-PEI/Beclin-siRNA下調多重抗藥性蛋白而提高紫杉醇對肺癌細胞的效益
CS-PEI/Beclin-siRNA下調多重抗藥性蛋白而提高紫杉醇對肺癌細胞的效益
將聚乙烯亞胺(Polyethylenimine, PEI)修飾到硫酸化軟骨素(Chondroitin sulfate, CS)，形成一個PEI 接枝CS 之共聚物(簡稱CS-PEI)。PEI是目前市售的一個黃金商品，具高轉染效能的非病毒式基因載體，但是PEI 所擁有的高正電荷密度也對細胞造成很高的毒性。本研究利用低分子量的PEI 接枝到天然的多醣體CS之側鏈上，一來解決臨床未來使用高分子量的PEI對細胞造成的毒性問題；二來以螞蟻雄兵之優勢，利用接掛多條低分子量PEI，使其仍然保留對基因的高轉染率。由於CD44常被發現於腫瘤細胞上，而CS可以辨識細胞膜表面的CD44受體，利用CS-PEI做為基因藥物載體，將增加CS-PEI攜帶之任何基因藥物對細胞表面過度表現CD44的腫瘤或腫瘤幹細胞的靶向性，經由CD44介導的胞飲作用，有效地進入細胞內，提高基因藥物在癌細胞的轉染效能。本研究即以這個低毒性、高轉染的CS-PEI來做為調控自噬作用的Beclin siRNA基因遞送載體；並以紫杉醇(Paclitaxel, PTX)抗藥性的腫瘤細胞株做為細胞模型(如肺癌NCI-H23-TXR)，期待藉由抑制細胞的自噬作用，來恢復PTX對腫瘤組織的抑制生長效果。結果發現Beclin siRNA不但抑制Beclin蛋白表現，同時也抑制數個多重抗藥性蛋白的表現，進而促使PTX有效地抑制腫瘤組織的增生。本研究同時也建立一套利用斑馬魚植入腫瘤，快速篩選藥物抑制腫瘤生長的測試平台，未來此平台可延伸至其他腫瘤植入的動物模型。除此，本研究使用的基因載體材料CS-PEI已經獲准台灣專利(I434934)及美國專利(US 8,445,025 B2; US 8,716,399 B2; US 9,050,362 B2)，顯見此材料開發具有相當優越的市場開拓新契機。
本校主要研究者之簡介:
本校醫藥暨應用化學系王麗芳教授領導的團隊，集結醫藥化學、生物醫學、奈米醫學、生醫工程、臨床醫學等跨領域專業人才，從化學合成、基因載體的設計、MDR細胞株的建立、自噬作用基因表現、班馬魚腫瘤植入之建立等等，是一個成功的跨系、院、校的成功例子。研究經費由科技部與高雄醫學大學共同支持。
研究聯繫Email: lfwang@kmu.edu.tw; cchiu@kmu.edu.tw
期刊出處: Molecular Therapy: Nucleic Acids, 2019, 17(9), 477-490.
期刊線上參閱網址:
https://www.cell.com/action/showPdf?pii=S2162-2531(19)30180-5
CS-PEI/Beclin-siRNA downregulate multidrug resistance proteins and increase paclitaxel therapeutic efficacy against NSCLC
CS-PEI/Beclin-siRNA downregulate multidrug resistance proteins and increase paclitaxel therapeutic efficacy against NSCLC
Polyethyleneimine (PEI) is one of leading cationic polymer for gene delivery because of its high transfection efficiency. However, PEI induced high cytotoxicity due to the positively charged surface characteristics. Herein, we grafted PEI onto chondroitin sulfate (CS) to yield CS-PEI which not only showed transgene efficiency, but reduced PEI cytotoxicity. CD44 is overexpressed in many solid tumor cells. It has been found that CS has the potential to be internalized into cells via CD44-mediated endocytosis. Thus, using CS-PEI as a gene drug delivery system, actively targeting to CD44-overexpressing cancer cells is an ideal approach to enhance transgene efficiency. Multidrug resistance (MDR) is the major obstacle in limitation of therapeutic efficacy of Paclitaxel (PTX). PTX-resistant non-small-cell lung cancer cell line (NCI-H23-TXR) was established and CS-PEI/Beclin-small interfering RNA (siRNA) was constructed to restore sensitivity of PTX against NCI-H23-TXR. Results revealed that knockdown of Beclin simultaneously inhibited MDR-related proteins, and renewed the sensitivity of PTX against NCI-H23-TXR. In vivo study showed that pre-transfection with CS-PEI/Beclin-siRNA followed by PTX treatment decreased the tumor size in NCI-H23-TXR zebrafish xenografts. (Taiwan Patent No. I434934; United States Patent US 8,445,025 B2; US 8,716,399 B2; US 9,050,362 B2)
Main researcher Intro.
Dr. Li-Fang Wang, a professor of Department of Medicinal and Applied Chemistry at Kaohsiung Medical University.
Author Email: lfwang@kmu.edu.tw ; cchiu@kmu.edu.tw
Paper cited from: Molecular Therapy: Nucleic Acids, 2019, 17(9), 477-490.
Paper online website:
https://www.cell.com/action/showPdf?pii=S2162-2531(19)30180-5
