Kitano Hisataka, Mamiya Atsushi, Ishikawa Tomomi, Egoshi Kayo, Kokubun Shinichiro, Hidai Chiaki
Division of Dental Surgery, School of Medicine, Nihon University, Tokyo, Japan.
Division of Physiology, Department of Biomedical Science, School of Medicine, Nihon University, Tokyo, Japan.
Onco Targets Ther. 2016 Jan 25;9:503-16. doi: 10.2147/OTT.S90801. eCollection 2016.
Cancer gene therapy using nonviral vectors is useful for long periods of treatment because such vectors are both safe and inexpensive, and thus can be used repeatedly. It has been reported that gene therapy with an E3C1 fragment of Del1 in a mouse explanted tumor model improved prognosis. The present study aimed to analyze the long-term effects of repeated non-viral gene transfer of E3C1. Mice with explanted tumors of SCCKN cells, a human squamous carcinoma, were treated with a plasmid encoding E3C1. Plasmids were injected locally every week using a transfection reagent. Control mice treated with mock DNA started to be euthanized on day 18, because the tumors had grown to over 15% of the body weight, and all of them had died by day 43. On the other hand, the tumors in two of ten mice treated with E3C1 had disappeared. The other eight mice started to be euthanized on day 46 and eight of ten mice had been euthanized by day 197. After 18 days of therapy, the tumor volume of control mice was 2,804±829 mm(3) and that of the E3C1 mice was 197±159 mm(3). Histochemical studies showed enhanced apoptosis in the E3C1-treated tumors, as compared with controls. Changes in cell morphology and decreased polymerized actin induced by E3C1 indicated disturbed cell adhesion to the matrix. In in vitro studies of SCCKN cells, prolonged administration of an E3C1 recombinant protein to cultured cells reduced adhesion-independent growth of cancer cells, as compared with control cells. These data suggest that E3C1 treatment induces anoikis.
使用非病毒载体的癌症基因治疗对于长期治疗很有用,因为这类载体既安全又廉价,因此可以反复使用。据报道,在小鼠移植瘤模型中,用Del1的E3C1片段进行基因治疗可改善预后。本研究旨在分析E3C1重复非病毒基因转移的长期效果。用人鳞状癌SCCKN细胞的移植瘤小鼠,用编码E3C1的质粒进行治疗。每周使用转染试剂将质粒局部注射。用空载体DNA处理的对照小鼠在第18天开始实施安乐死,因为肿瘤已生长到超过体重的15%,并且所有对照小鼠在第43天均死亡。另一方面,用E3C1治疗的十只小鼠中有两只的肿瘤消失了。另外八只小鼠在第46天开始实施安乐死,十只小鼠中有八只在第197天被安乐死。治疗18天后,对照小鼠的肿瘤体积为2,804±829 mm³,E3C1处理小鼠的肿瘤体积为197±159 mm³。组织化学研究表明,与对照相比,E3C1处理的肿瘤中凋亡增强。E3C1诱导的细胞形态变化和聚合肌动蛋白减少表明细胞与基质的粘附受到干扰。在SCCKN细胞的体外研究中,与对照细胞相比,向培养细胞长期施用E3C1重组蛋白可降低癌细胞的非粘附性生长。这些数据表明E3C1治疗可诱导失巢凋亡。