论文部分内容阅读
目的探讨靶向血管内皮细胞生长因子(VEGF)基因的小干扰RNA(siRNA)联合负载肿瘤抗原的树突状细胞(DC)致敏的细胞毒性T淋巴细胞(CTL)的联合抗瘤作用。方法体外应用靶向VEGF基因最佳转染浓度(100nmol/L,增加浓度不再提高沉默作用)的siRNA联合负载人乳腺癌细胞(MCF-7细胞)抗原的DC介导的CTL(siRNAVEGF+CTL组)共同作用于MCF-7细胞,同时设立空白对照组和空白+CTL组(只加无血清无抗生素培养基);脂质体组和脂质体+CTL组(只加LipofectamineTM2000);siRNAVEGF-组(100nmol/LsiRNA);siRNASCR组和siR-NASCR+CTL组(100nmol/LsiRNASCR),每组做6个平行孔,四甲基偶氮唑蓝(MTT)法检测siRNAVEGF+CTL组和各对照组肿瘤杀伤活性(n=6),Hoechst33258核染色观察细胞的凋亡(n=3)。结果siRNAVEGF+CTL组、siR-NAVEGF-组siRNASCRCTL组肿瘤杀伤活性分别为99.37%,51.17%和43.94%,siRNAVEGF+CTL组与对照组比较可明显杀伤肿瘤细胞,瘤细胞几乎完全溶解,Hoechst33258显示细胞核呈明显的细胞凋亡改变。结论体外实验显示靶向VEGF的siRNA联合负载肿瘤抗原DC致敏的CTL能有效抑制乳腺癌MCF-7细胞的生长,二者的联合应用抗瘤效果显著。
Objective To investigate the combined anti-tumor effect of small interfering RNA (siRNA) targeting vascular endothelial growth factor (VEGF) gene and dendritic cell (DC) -sensitive cytotoxic T lymphocytes (CTL) loaded with tumor antigen. Methods DC-mediated CTL (siRNAVEGF + CTL) combined with siRNA targeting human breast cancer cell (MCF-7 cell) antigens targeting siRNA targeting the optimal transfection concentration of VEGF gene (100 nmol / L with no further increase of silencing) Group), MCF-7 cells were also established. At the same time, blank control group and blank + CTL group (only serum-free antibiotic-free medium) were established; liposome group and liposome + CTL group (only LipofectamineTM2000); siRNAVEGF- (100nmol / LsiRNA), siRNASCR group and siR-NASCR + CTL group (100nmol / LsiRNASCR), 6 parallel wells in each group, siRNAVEGF + CTL group and each control group were detected by MTT assay Tumor killing activity (n = 6), apoptosis of cells were observed by Hoechst33258 nuclear staining (n = 3). Results The tumor-killing activity in siRNAVEGF + CTL group and siR-NAVEGF-group siRNASCRCTL group were 99.37%, 51.17% and 43.94% respectively. Compared with the control group, siRNAVEGF + CTL group could obviously kill tumor cells and the tumor cells almost completely dissolved. Hoechst33258 showed that the nucleus Significant changes in apoptosis. Conclusion In vitro experiments showed that siRNA targeting VEGF combined with CTL loaded with tumor antigen DC could effectively inhibit the growth of breast cancer MCF-7 cells. The combination of the two showed significant antitumor effects.