CAJ | 학술논문

目的:制备携带人基质金属蛋白酶组织抑制因子-1(tissue inhibitors of metalloproteinase-1,TIMP-1)的重组腺病毒乳酸聚乙烯醇(poly-DL-lactide-poly,PELA)微球,探讨其对HepG2肝癌细胞增殖的影响.方法:采用溶剂挥发法双乳液体系,以可降解的生物材料PELA包被携带TIMP-1基因的重组腺病毒制成微球,测定其粒径、载病毒量、包封率及释放规律.重组腺病毒微球感染HepG2细胞,荧光显微镜观测感染效率,透射电镜观测超微结构,半定量RT-PCR检测TIMP-1 mRNA表达;MTT法检测HepG2细胞增殖.结果:成功构建包载TIMP-1重组腺病毒的PELA微球,直径约1.965 μm,包封率为60%,载病毒率为10.5×10~8efu/mg,在120 h内释放病毒量接近60%,总的释放时间长于240 h.空白微球无毒性PELA病毒微球感染HepG2细胞后,细胞稳定表达TIMP-1 mRNA;对HepG2细胞的增殖有明显抑制作用,抑制率表达47%.结论:包载TIMP-1重组腺病毒的PELA微球可抑制肝癌HepG2细胞的增殖,为化学高分子载体运载基因治疗肝癌提供了实验依据.
목적:제비휴대인기질금속단백매조직억제인자-1(tissue inhibitors of metalloproteinase-1,TIMP-1)적중조선병독유산취을희순(poly-DL-lactide-poly,PELA)미구,탐토기대HepG2간암세포증식적영향.방법:채용용제휘발법쌍유액체계,이가강해적생물재료PELA포피휴대TIMP-1기인적중조선병독제성미구,측정기립경、재병독량、포봉솔급석방규률.중조선병독미구감염HepG2세포,형광현미경관측감염효솔,투사전경관측초미결구,반정량RT-PCR검측TIMP-1 mRNA표체;MTT법검측HepG2세포증식.결과:성공구건포재TIMP-1중조선병독적PELA미구,직경약1.965 μm,포봉솔위60%,재병독솔위10.5×10~8efu/mg,재120 h내석방병독량접근60%,총적석방시간장우240 h.공백미구무독성PELA병독미구감염HepG2세포후,세포은정표체TIMP-1 mRNA;대HepG2세포적증식유명현억제작용,억제솔표체47%.결론:포재TIMP-1중조선병독적PELA미구가억제간암HepG2세포적증식,위화학고분자재체운재기인치료간암제공료실험의거.
Objective: Toprepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) adenovirus, and to investigate their effects on the proliferation of hepatocellular carcinoma HepG2 cells. Methods: The microsphere was constructed by encapsulating recombinant adenovirus containing TIMP-1 in biodegradable PELA. The diameter of the microsphere, quantity of virus encapsulated, loading rate, and releasing kinetics were measured. HepG2 cells were infected with the microspheres; the infection efficiency was examined by fluorescent microscope; and the ultrastructure was observed by TEM. The expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR, and the proliferation of HepG2 cells was detected by MTT assay. Results: The microsphere encapsulating recombinant TIMP-1 adenovirus was successfully constructed, with its diameter, entrapment efficiency, and virus loading rate being 1.965, 60.0%, and 10.5×10~8/mg, respectively. About 60% of the viruses were released within 120 h, and the total releasing time was longer than 240 h. Infection with rAdTIMP-1 PELA microsphere efficiently induced TIMP-1 expression in HepG2 cells, and significantly inhibited the proliferation of HepG2 cells, with the inhibitory rate being 47%. Conclusion: PELA microsphere encapsulating recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for the combining macromolecular chemistry and gene therapy for treatment of hepatocellular carcinoma.