CAJ | 학술논문

目的观察利用阳性纳米脂质体介导的基因治疗联合免疫治疗对结肠癌细胞的治疗作用.方法应用基因克隆技术分别构建含有绿色荧光蛋白报告基因的重组质粒(pEGFP),并依据静电吸附原理制备携带基因的阳离子纳米脂质体,再将转染阳性的树突状细胞(DC)移植作用于结肠癌细胞.结果成功构建含有绿色荧光蛋白报告基因的重组质粒,并得到酶切鉴定和序列测定证实.携带酪氨酸激酶受体3 配体(FL)基因的阳离子纳米脂质体成功转染DC,发现基因在细胞内得到很好表达,且随着培养时间的增加,蛋白表达量有所增加,72h时表达量最高.基因移植后发现对照组肿瘤体积增长最快,FL基因组肿瘤体积增长较慢；FL基因组肿瘤细胞凋亡率和死亡率明显高于对照组(P＜0.05)；而pEGFP组和对照组差异无统计学意义(P＞0.05).结论阳离子纳米脂质体作为基因载体具有一定优越性.基因治疗联合免疫治疗在结肠癌治疗中具有重要意义.
목적 관찰이용양성납미지질체개도적기인치료연합면역치료대결장암세포적치료작용.방법 응용기인극륭기술분별구건함유록색형광단백보고기인적중조질립(pEGFP),병의거정전흡부원리제비휴대기인적양리자납미지질체,재장전염양성적수돌상세포(DC)이식작용우결장암세포.결과 성공구건함유록색형광단백보고기인적중조질립,병득도매절감정화서렬측정증실.휴대락안산격매수체3 배체(FL)기인적양리자납미지질체성공전염DC,발현기인재세포내득도흔호표체,차수착배양시간적증가,단백표체량유소증가,72h시표체량최고.기인이식후발현대조조종류체적증장최쾌,FL기인조종류체적증장교만；FL기인조종류세포조망솔화사망솔명현고우대조조(P＜0.05)；이pEGFP조화대조조차이무통계학의의(P＞0.05).결론 양리자납미지질체작위기인재체구유일정우월성.기인치료연합면역치료재결장암치료중구유중요의의.
Objective To investigate the therapeutic effects of cationic nano-liposome-mediated gene therapy combined with immunotherapy for colon cancer.Methods Recombinant plasmids containing green and red fluorescent protein reporter genes were constructed using gene cloning methods.Gene-carrying cationic nano-liposomes were prepared based on the electrostatic adherence principle and then transfected into dendritic cells (DC) that were transplanted into colon cancer cells.Results Recombinant plasmids containing green or red fluorescent protein reporter genes were successfully constructed using gene cloning methods and confirmed by restriction enzyme digestion and sequencing.Gene-carrying cationic nano-liposomes were transfected into colon cancer cells,and gene expression was detected.The apoptosis and cell death was observed in the tyrosine kinase receptor 3 ligand (FL) group,and the lowest level was detected in the control group.The results revealed significantly higher level of apoptisis and lower level of proliferation in the FL group than in the control groups ( P ＜ 0.05 ).However,the lowest level of tumor cell apoptisis and the highest level of cell proliferation were observed in the control group.Conclusion Cationic nano-liposomes have the advantages of being gene carriers.Gene therapy combined with immunotherapy may be important for colon cancer.