CAJ | 학술논문

目的构建Trastuzmab F(ab’)2修饰的阿霉素免疫脂质体并观察其对体外生长的人乳腺癌MCF-7细胞的杀伤作用.方法抗体交联法制备Trastuzmab F(ab’)2段修饰的阿霉素免疫脂质体；透射电子显微镜观察免疫脂质体的形态及粒径分布；采用荧光标记法测定免疫脂质体活性；噻唑蓝(MTT)法检测MCF-7细胞在不同药物浓度下培养8、16、24、32 h后的增殖差异；流式细胞术检测200 mg/L药物浓度下培养32 h后的各组细胞凋亡.结果成功构建了Trastuzmab F(ab’)2修饰的阿霉素免疫脂质体；所得免疫脂质体平均粒径为192 nm,粒径小于200 nm者占92.20％；投药10、30、50 min时摄入阿霉素免疫脂质体及阿霉素脂质体的细胞比例分别为(19.39±3.33)％、(57.36 ±4.12)％、(89.72±5.88)％和(10.17±4.05)％、(25.15±3.77)％、(38.86±6.24)％(P＜0.05)；阿霉素免疫脂质体对MCF-7细胞增殖的抑制作用强于阿霉素脂质体(F=1 256.225,P＜0.01),且该抑制作用具有时间(F=2 145.325,P＜ 0.01)和浓度(F=1 287.472,P＜0.01)的依赖性；相同时间内,MCF-7细胞对阿霉素免疫脂质体的摄取要明显高于对照组(P＜0.05)；TrastuzmabF(ab,)2修饰的阿霉素免疫脂质体对MCF-7细胞增殖具有一定的抑制作用(P＜0.01),且该抑制作用具有时间和浓度的依赖性(P＜0.01)；转染32 h后,阿霉素免疫脂质体组的细胞凋亡率[(35.22±3.52)％]高于对照组[(10.31±4.62)％,P＜ 0.05].结论成功构建TrastuzmabF(ab,)2修饰的阿霉素免疫脂质体,其对体外生长的乳腺癌细胞具有较强的杀伤作用.
목적 구건Trastuzmab F(ab’)2수식적아매소면역지질체병관찰기대체외생장적인유선암MCF-7세포적살상작용.방법 항체교련법제비Trastuzmab F(ab’)2단수식적아매소면역지질체；투사전자현미경관찰면역지질체적형태급립경분포；채용형광표기법측정면역지질체활성；새서람(MTT)법검측MCF-7세포재불동약물농도하배양8、16、24、32 h후적증식차이；류식세포술검측200 mg/L약물농도하배양32 h후적각조세포조망.결과 성공구건료Trastuzmab F(ab’)2수식적아매소면역지질체；소득면역지질체평균립경위192 nm,립경소우200 nm자점92.20％；투약10、30、50 min시섭입아매소면역지질체급아매소지질체적세포비례분별위(19.39±3.33)％、(57.36 ±4.12)％、(89.72±5.88)％화(10.17±4.05)％、(25.15±3.77)％、(38.86±6.24)％(P＜0.05)；아매소면역지질체대MCF-7세포증식적억제작용강우아매소지질체(F=1 256.225,P＜0.01),차해억제작용구유시간(F=2 145.325,P＜ 0.01)화농도(F=1 287.472,P＜0.01)적의뢰성；상동시간내,MCF-7세포대아매소면역지질체적섭취요명현고우대조조(P＜0.05)；TrastuzmabF(ab,)2수식적아매소면역지질체대MCF-7세포증식구유일정적억제작용(P＜0.01),차해억제작용구유시간화농도적의뢰성(P＜0.01)；전염32 h후,아매소면역지질체조적세포조망솔[(35.22±3.52)％]고우대조조[(10.31±4.62)％,P＜ 0.05].결론 성공구건TrastuzmabF(ab,)2수식적아매소면역지질체,기대체외생장적유선암세포구유교강적살상작용.
Objective To construct the immunoliposomal adriamycin tagged with trastuzumab F (ab') 2 and to investigate its killing effect on human breast cancer cells in vitro.Methods The tastuzmab F(ab')2 and the liposomal adriamycin were linked together by antibody crosslinking method.The morphology of immunoliposomes was observed by transmission electron microscopy (TEM) and their size distribution was measured by graphics software.The immune activity of immunoliposomes was evaluated using the fluorescence microscopy after the fluorescent immunoliposomes or lipoosomes were incubated together with MCF-7 cells.Inhibitory effect of the cell growth at different drug concentrations was measured using methyl thiazol tetrazolium (MTT) 8,16,24 and 32 h after cell culture.The apoptosis was examined by using flow cytometry 32 h after cell culture.Results The immunoliposomal adriamycin tagged with trastuzumab F(ab')2 that was constructed in vitro has an average diameter of 192 nm,and that the diameters under 200 nm accounted for 92.20％ of the total.The immunoliposome was superior to liposome at the identification capability to MCF-7 cells,the proportions of the cells which intake drug were (19.39 ±3.33)％,(57.36 ±4.12)％,(89.72 ±-5.88)％ and (10.17 ±4.05)％,(25.15 ± 3.77)％,(38.86 ±6.24)％ (P＜0.05) in two groups respectively after 10,30 and 50 min cell cultrue.The immunoliposome could inhibit the proliferation of MCF-7 cells in a dose (F =1 287.472,P ＜ 0.01) and time (F =2 145.325,P＜ 0.01)dependent manner; Apoptotic index of MCF-7 cells in immunoliposome group (35.22 ± 3.52) ％ was obviously higher than that in the control groups (10.31 ± 4.62) ％ 32 h after transfection.(P ＜ 0.05).Conclusion The immunoliposomal adriamycin tagged with trastuzumab F (ab') 2 was constructed successfully and it can inhibit the proliferation of kill the MCF-7 cells effectively in vitro.