CAJ | 학술논문

目的观察电穿孔介导免疫调节因子及血管生成抑制因子转移治疗脉络膜黑色素瘤(CM)的效果.方法采用编码小鼠免疫调节因子白介素(IL)2、IL12、血管内皮细胞生长因子(VEGF)可溶性受体(sFLK-1)、反义血管内皮细胞生长因子(antiVEGF121)及血管生成素1可溶性受体(ExTek)的真核表达质粒pNGVL-mIL2、pNGVL-mIL12-、pCI-sFLK-1、pCR3.1-antiVEGF121、pCI-ExTek分别转染人胚肾细胞和人CM.采用酶联免疫吸附测定法(ELISA)、蛋白免疫印迹法(Western blot)检测mIL2、mIL12、sFLK-1、VEGF和ExTek因子在转染细胞中的表达.建立裸小鼠肿瘤模型并随机分为4组,分别将体积30μl的0.9%NaCl、pNGVL、antiVEGF121+sFLK-1+ExTek、mIL2+mIL12注射入肿瘤内.采用高电压短脉冲方式进行电击.治疗后7、14、21、28、35、42 d测量各组裸小鼠肿瘤体积,计算抑瘤率.结果 ELISA及Western blot检测显示,mIL2、mIL12、sFLK-1和ExTek因子均能在转染细胞中有效表达;而VEGF在转染细胞中表达明显受抑制.电穿孔基因治疗后,mIL2+mIL12组肿瘤几乎完全消失,抑瘤率为97.33%;antiVEGF121+sFLK-1+ExTek组及pNGVL组肿瘤持续长大,抑瘤率分别为53.33%和36.33%.结论电穿孔介导免疫调节因子转移可有效抑制CM生长;介导血管生成抑制因子转移不能抑制CM生长.
목적 관찰전천공개도면역조절인자급혈관생성억제인자전이치료맥락막흑색소류(CM)적효과.방법 채용편마소서면역조절인자백개소(IL)2、IL12、혈관내피세포생장인자(VEGF)가용성수체(sFLK-1)、반의혈관내피세포생장인자(antiVEGF121)급혈관생성소1가용성수체(ExTek)적진핵표체질립pNGVL-mIL2、pNGVL-mIL12-、pCI-sFLK-1、pCR3.1-antiVEGF121、pCI-ExTek분별전염인배신세포화인CM.채용매련면역흡부측정법(ELISA)、단백면역인적법(Western blot)검측mIL2、mIL12、sFLK-1、VEGF화ExTek인자재전염세포중적표체.건립라소서종류모형병수궤분위4조,분별장체적30μl적0.9%NaCl、pNGVL、antiVEGF121+sFLK-1+ExTek、mIL2+mIL12주사입종류내.채용고전압단맥충방식진행전격.치료후7、14、21、28、35、42 d측량각조라소서종류체적,계산억류솔.결과 ELISA급Western blot검측현시,mIL2、mIL12、sFLK-1화ExTek인자균능재전염세포중유효표체;이VEGF재전염세포중표체명현수억제.전천공기인치료후,mIL2+mIL12조종류궤호완전소실,억류솔위97.33%;antiVEGF121+sFLK-1+ExTek조급pNGVL조종류지속장대,억류솔분별위53.33%화36.33%.결론 전천공개도면역조절인자전이가유효억제CM생장;개도혈관생성억제인자전이불능억제CM생장.
Objective To observe the effects of immunologic cytokines or anti-angiogenesis gene transfer mediated by electroporation for choroidal melanoma cells. Methods The human embryo kidney cells and malignant choroidal melanoma cells were transfected with plasmids pNGVL-mIL2, pNGVL-mIL12, pCI-sFLK-1, pCR3. 1-antiVEGF121. pCI-ExTek. Then the expression of mIL2, mIL12, sFLK-1,VEGF and ExTek were detected by enzyme-linked immunosorbentassay (ELISA) and Western blot. Nude mice models of malignant choroidal melanoma were established and they were divided into four groups randomly. Each group was treated with 30 μl of 0. 9% NaCl, 30 μg pNGVL, 30 μμg antiVEGF121 + sFLK-1 +ExTek and 30 μg mIL2+mIL12 respectively by electroporation. Seven, 14, 21, 28, 35 and 42 days after treatment, the tumor volumes were measured to calculate the tumor inhibition rate. Results ELISA and Western blot showed that mIL2, mIL12, sFLK-1 and ExTek were expressed after electroporation, VEGF expression was decreased remarkably. After treatment, the tumors of mIL2 + mIL12 group were greatly inhibited with a tumor inhibition rate of 97.33%, while the tumors of antiVEGF121 + sFLK-1 + ExTek and pNGVL group were partially inhibited with tumor inhibition rates of 53. 33% and 36. 33% respectively.Conclusions Immunologic cytokines transfer mediated by electroporation can inhibit the growth of melanoma,but anti-angiogenesis only have a mild effects.