Cell-based protein delivery system for the inhibition of the growth of pancreatic cancer: NK4 gene-transduced oral mucosal epithelial cell sheet.
Cell-based protein delivery system for the inhibition of the growth of pancreatic cancer: NK4 gene-transduced oral mucosal epithelial cell sheet.
复制标题
用于抑制胰腺癌生长的基于细胞的蛋白质递送系统：NK4基因转导的口腔粘膜上皮细胞片。
作者：
T. Manabe;K. Mizumoto;E. Nagai;Kunio Matsumoto;Toshikazu Nakamura;T. Nukiwa;Masao Tanaka;T. Matsuda
目的
胰腺癌手术切除是唯一的治疗方法，但术后局部复发率高，预后极差。本研究旨在开发一种新的抑制残留癌细胞生长的治疗工具，该工具基于局部递送NK4(肝细胞生长因子拮抗剂)，该NK4(肝细胞生长因子拮抗剂)由NK4基因转导的口腔黏膜上皮细胞(Omec)片(NK4-Sheet)分泌，并附着在切除的表面上。
实验设计
按照3T3饲养层技术采集和培养OMEC，将其种植在胶原网-覆盖层的可生物降解的VICRYL网上，制成OMEC片材。重组腺病毒(Ad-NK4)介导NK4基因转导。检测了OMECs对基于细胞的NK4输送的适用性。建立了一种裸鼠实验模型，以确定NK4-Sheet对肿瘤生长和血管生成的影响。
结果
Ad-NK4转导的OMECs分泌的NK4抑制MRC-5诱导的胰腺癌细胞株的侵袭。异位移植的转导基因的OMECs存活10天后逐渐减少。NK4片层抑制了体内血管生成和肿瘤生长。
结论
自体Omec被发现适合于这一目的，因为它不分泌肝细胞生长因子，易于从患者身上获取，具有合理的高增殖潜力，并且没有免疫反应。尽管正在开发的NK4-Sheet表现出低水平和短时间的NK4分泌，但当该系统负载多层OMEC时，有望在临床情况下具有很大的靶向组织的蛋白质递送系统的潜力。
PURPOSE
Pancreatic resection for pancreatic cancer is the only curative modality, but the high incidence of local recurrence after surgery results in a very poor prognosis. This study aims to develop a new therapeutic tool that could inhibit the growth of remnant cancer cells, which is based on local delivery of NK4 (hepatocyte growth factor antagonist) secreted from an NK4 gene-transduced oral mucosal epithelial cell (OMEC) sheet (NK4-sheet), which is adhered to the resected surface.
EXPERIMENTAL DESIGN
OMECs, harvested and cultured according to 3T3 feeder layer technique, were seeded on a collagen mesh-overlayered, biodegradable VICRYL mesh to produce an OMEC sheet. NK4 gene transduction was mediated by recombinant adenovirus (Ad-NK4). Applicability of OMECs for cell-based NK4 delivery was examined. An experimental model using nude mice was established to determine the effect of an NK4-sheet on both tumor growth and angiogenesis.
RESULTS
NK4 secreted from Ad-NK4-transduced OMECs suppressed MRC-5-induced invasion of pancreatic cancer cell lines. Heterotopically implanted gene-transduced OMECs remained for >/==" BORDER="0">10 days while gradually decreasing. NK4-sheets inhibited both angiogenesis and tumor growth in vivo.
CONCLUSION
Autologous OMEC was found to be suited to this purpose because of no secretion of hepatocyte growth factor, ease in harvesting from a patient, reasonably high proliferation potential, and no immune reaction. Although NK4-sheets under development exhibited a low level and short period of NK4 secretion, it is expected that this system may have a great potentiality of protein delivery system to target tissue at clinical situations when it is loaded with multilayered OMECs.