摘要
目的探讨HIFU联合脂质体介导p53基因转染胰腺癌细胞的可行性以及转染后对细胞生物学行为的影响。方法实验分为5组:I组为单纯细胞组,II组为HIFU+质粒组,III组为HIFU+脂质体+质粒组,IV组为脂质体+质粒组,V组为单纯质粒组。每组予以相应处理后,采用RT-PCR法检测各组细胞p53 mRNA表达情况,MTT法及流式细胞术检测细胞增殖、凋亡的情况。结果荧光显微镜下见Ⅲ组有较多绿色荧光蛋白表达的细胞(转染率为10.6%),II组及IV组有少量的荧光细胞(转染率分别为1.2%,4.8%);V组仅有极少的荧光细胞(0.3%);I组无荧光表达;III组和IV组经RTPCR法均检测到p53mRNA表达,且两组灰度值差异有显著性意义(P<0.05);MTT结果显示II,III,IV组细胞均有明显的生长抑制作用,而其中III组的增殖抑制作用最为明显,与其它各组比较差异具有显著性意义(P<0.05);Ⅲ组细胞凋亡率为(14.2±1.55)%,与其它4组比较,凋亡率明显增高(P<0.05)。结论 HIFU联合脂质体可提高p53质粒转染率;p53质粒转染后促凋亡作用增强,肿瘤细胞生长进一步受到抑制。
Objective To explore the feasibility of p53 transfected into pancreatic cancer cells mediated by high intensity focused ultrasound combined with liposome and the effect of the transfection on cell biological behaviour. Methods Pancreatic cancer cells were divided into 5 groups： simple cell group （Group）, HIFU-plasmid group （Group II）, HIFU-liposome-plasmid group （Group Ⅲ）, liposome-plasmid group （ Group Ⅳ） , simple plasmid group （ Group Ⅴ）. After proper treatment for the cells in the five groups, the mRNA expression of p53 in all the cells was detected with RT-PCR, and the proliferation and apoptosis of cells were detected with MTT assay and flow cytometry. Results With fluorescence microscopy, many cells with expression of green fluorescent protein were found in group III （with a transfection efficiency of 10. 6% ） , a few in Group II and Group IV （with a transfection efficiency of 1. 2% and 4. 8% respectively） , few in Group V （with a transfection efficiency of 0. 3% ） , and none in Group I. The mRNA expression of p53 was found in Group III and Group IV, with significant difference in the grey level between the two groups （ P〈0. 05 ）. The results of MTT showed that the significant cell growth inhibition existed in Group II, Group IIl and G^oup IV, and the inhibition in Group Ⅱ appeared the most notably with significant difference from other groups （P〈0. 05 ）. The apoptosis rate in Group IIl was （ 14. 2＋ 1.55） %, significantly higher than that in other groups （P〈0. 05）. Conclusions The transfection efficiency of p53 into pancreatic cells can be enhanced by HIFU combined with liposome. The transfection of p53 promotes cell apoptosis, further inhibiting the growth of tumor cells.
出处
《辽宁医学院学报》
CAS
2016年第4期13-16,I0003,共5页
Journal of Liaoning Medical University (LNMU) Bimonthly
基金
2012年江苏省徐州市科技计划项目
编号:XM12B042
