携带4-1BBL基因的胃癌细胞总RNA转染树突状细胞疫苗的研究

Dendritic cell vaccine-transfected gastric cancer cell total RNA carrying 4-1BBL gene

目的 观察携带4-1BBL基因的胃癌细胞总RNA转染树突状细胞疫苗,在小鼠体内诱导的抗肿瘤效应以及对小鼠免疫力的影响.方法 用脂质体法把pMKITneo/4-1 BBL质粒导入小鼠胃癌细胞MFC内,再提取此细胞的总RNA并转染至树突状细胞内制备MFC/4-1BBL/DC疫苗;把MFC细胞注射于20只615小鼠皮下建立荷瘤小鼠胃癌模型,并随机分为对照组、DC组、MFC/DC组和MFC/4-1BBL/DC组,每组5只;在MFC细胞接种后第7、14天给予相应DC疫苗治疗,于肿瘤细胞接种后第21天,处死实验动物,测量瘤重、测定肿瘤细胞的凋亡率及外周血的CD4＋、CD8＋T细胞和NK细胞的含量.结果 MFC/4-1BBL/DC组小鼠平均瘤重（2.06±0.39）g显著低于对照组（3.82±0.57）g、DC组（3.63±0.51）g、MFC/DC组（2.67±0.32）g（P＜0.05）;且MFC/4-1BBL/DC组肿瘤细胞凋亡率（28.58±2.84）%明显高于MFC/DC组（25.03±1.88）%、DC组（20.66±2.71）%及对照组（19.09±2.73）%（P＜0.05）;MFC/4-1BBI/DC组小鼠外周血CID4＋T、NK细胞数明显高于对照组和其他治疗组（P＜0.05）.结论 携带4-1BBL基因的胃癌细胞总RNA转染树突状细胞疫苗能够提高荷瘤机体的免疫能力、抑制肿瘤细胞的生长、促进肿瘤细胞的凋亡.

Objective To observe the induced anti-tumor effects and immune state in vivo by the transfection of dendritic cell vaccine into the total RNA of gastric cancer cells carrying 4-1BBL gene.Methods The liposome-mediated pMKITneo/4-1BBL gene was inserted into the MFC gastric cancer cells,and the cell total RNA was extracted and transfected into dendritic cells to make the MFC/4-1BBL/DC vaccine.After MFC cells were injected into the 615 mouse to establish tumor-bearing mouse model,and those 20 mice were randomly divided into control group,DC group,MFC/DC group,MFC/4-1BBL/DC group.The dendritic cell vaccine was subcutaneously injected on the day 7 and 14 after inoculation of tumor cells,and on the day 21 animals were killed and tumor weight was measured,tumor cell apoptosis rate was assayed and the contents of CD4 ＋,CD8 ＋ T cells and NK cells in peripheral blood were analyzed.Results The mean tumor weight in MFC/4-1BBL/DC group （2.06 ±0.39） g was significantly less than that in control group （3.82 ±0.57） g,DC group （3.63 ±0.51 ） g,MFC/DC group （2.67 ±0.32） g （P 〈0.05）.Moreover the apoptosis rate in MFC/4-1BBL/DC group （28.58 ± 2.84 ） % was significantly higher than that in MFC/DC group （25.03 ± 1.88）%,DC group （20.66 ±2.71 ）% and control group （ 19.09 ±2.73 ）% （P 〈0.05）.The percentage of CD4 ＋ T,NK cells in MFC/4-1BBL/DC group was significantly higher than in other groups （ P〈0.05）.Conclusion The transfection of dendritic cell vaccine into the total RNA of gastric cancer cells carrying 4-1BBL gene can increase immunity of the bearing tumor mice,inhabit tumor growth and promote tumor cell apoptosis.