CAJ | 학술논문

目的研究跨膜型血型A抗原模拟肽疫苗诱导恶性黑色素瘤细胞B16凋亡的作用.方法采用脂质体转染法分别将前期构建的模拟肽疫苗稳定转染B16细胞,加入终浓度为2.5 μg/ml、5 μg/ml、10 μg/ml及20 μg/ml的抗血型A单克隆抗体培养72 h,Annexin Ⅴ/PI双染法检测细胞凋亡率.结果 P/F-M-pIRES组的B16细胞经10 μg/ml抗血型A单克隆抗体诱导后凋亡率为74.74%,转染有模拟肽/Fas融合基因的P/F-M-pIRES组和P/F-pIRES组凋亡率明显高于未转染的M-pIRES组和pIRES组.析因设计的方差分析显示不同实验分组之间的差异主效应差异有统计学意义(F＝669.707,P＜0.01),不同浓度分组之间的主效应差异有统计学意义(F＝106.596,P＜0.01),不同实验分组不同浓度之间的交互效应差异也有统计学意义(F＝34.806,P＜0.01).结论跨膜型血型A抗原模拟肽疫苗可诱导黑色素瘤细胞B16凋亡,在黑色素瘤治疗中可能是一种有潜在价值的方法.
목적 연구과막형혈형A항원모의태역묘유도악성흑색소류세포B16조망적작용.방법 채용지질체전염법분별장전기구건적모의태역묘은정전염B16세포,가입종농도위2.5 μg/ml、5 μg/ml、10 μg/ml급20 μg/ml적항혈형A단극륭항체배양72 h,Annexin Ⅴ/PI쌍염법검측세포조망솔.결과 P/F-M-pIRES조적B16세포경10 μg/ml항혈형A단극륭항체유도후조망솔위74.74%,전염유모의태/Fas융합기인적P/F-M-pIRES조화P/F-pIRES조조망솔명현고우미전염적M-pIRES조화pIRES조.석인설계적방차분석현시불동실험분조지간적차이주효응차이유통계학의의(F＝669.707,P＜0.01),불동농도분조지간적주효응차이유통계학의의(F＝106.596,P＜0.01),불동실험분조불동농도지간적교호효응차이야유통계학의의(F＝34.806,P＜0.01).결론 과막형혈형A항원모의태역묘가유도흑색소류세포B16조망,재흑색소류치료중가능시일충유잠재개치적방법.
Objective To investigate the apoptotic effect of the transmembrane form vaccine of human blood group A mimotope on malignant melanoma cell line B16. Methods B16 cells were transfected with different recombinant plasmid through Lipofectamine 2000 and incubated with different concentration of monoclonal anti-A antibody at 2.5 μg/ml, 5 μg/ml,10 μg/ml and 20 μg/ml. Apoptosis rate of cells was determined with Annexin Ⅴ/PI double staining by flow cytometry. Results Apoptosis rate to P/F-M-pIRES group B16 cells was 74.74% when anti-A monoclonal antibody concentration was 10 μg/ml; apoptosis rate of plasmids carrying peptide/Fas fusion gene such as P/F-M-pIRES group and P/F-pIRES group were significantly higher than M-pIRES group and pIRES group. The apoptosis rate was statistically significantly different between different recombinated plasmid groups (F＝669.707,P＜0.01). The apoptosis rate was statistically significantly different between different antibody groups (F＝106.596,P＜0.01). The interaction between recombinated plasmid groups and antibody groups was statistically significant (F＝34.806,P＜0.01). Conclusions The transmembrane form vaccine of human blood group A mimotope could induce B16 cell apoptosis in vitro. This vaccine may be a promising candidate for potential malignant melanoma therapy.