分泌性内皮抑制素真核质粒治疗小鼠肝癌的初步研究

Therapeutic effect of secretive endostatin eukaryotic expressing plasmid on mouse hepatoma

目的 构建并表达分泌性内皮抑制素真核表达质粒,以此对肝癌进行基因治疗。方法 人工合成Ig κ信号肽序列,和内皮抑制素编码序列一起克隆入pcDNA3.1质粒。重组质粒转染上清液作用于ECV304内皮细胞,MTT法检测内皮细胞的增殖。局部注射重组质粒治疗接种在小鼠腿部肌肉内的H_(22)肝癌瘤株,疗程结束后解剖称取瘤重。结果 构建的内皮抑制素真核表达质粒转染上清液可以抑制内皮细胞的增殖抑制率为29.2％。经质粒裸DNA注射治疗后,治疗组瘤重[(1.34±0.96)g]比空载体组[(2.70±0.82)g]和生理盐水组[(3.73±1.41)g]明显减小(P<0.05)。结论 分泌性内皮抑制素真核表达质粒用于H_(22)肝癌的基因治疗有一定效果。

Objective To construct and express secretive endostatin eukaryotic plasmid for treatment of hepatoma. Methods Mouse Igk signal peptide sequence was synthesized and cloned into pcDNA3.1 with endostatin gene. The supernant of BHK-21 transfected with recombinant was used to culture ECV304. The proliferation of latter was evaluated by MTT assay. H22 was inoculated intramusclely, then naked DNA of endostatin plasmid was injected into the inoculation site. Tumors were dissected and weighted after treatments. All data was analyzed by SPSS 10.0. Results The supernant of BHK-21 transfected with recombinant can inhibit the proliferation of ECV304 by 29.2%. Tumor weight lighter after injected with naked pSecES (1.34 g ± 0.96 g) compared with naked pcDNA3.1 (2.70 g± 0.82 g) and saline (3.73 g±1.41 g). Conclusions The endostatin eukaryotic plasmid was constructed and it can be used for gene therapy on hepatoma.