转hGM-CSF基因HepG2细胞疫苗侵袭性和生长活性初步检测  

Initial Detection of Invasion and Growth Activity of hGM-CSF Gene Transfected HepG2 Cell Vaccine

作　　者：程元星[1] 段晓明[2] 曾治中[1] 黄璐[1] 贺修胜[3] 

机构地区：[1]南华大学研究生院,湖南衡阳421001 [2]长沙市第四医院消化内科,湖南长沙410000 [3]南华大学肿瘤研究所,湖南衡阳421001

出　　处：《现代生物医学进展》2011年第9期1621-1624,1604,共5页Progress in Modern Biomedicine

基　　金：湖南省卫生厅科研基金资助项目(B2005-178)

摘　　要：目的:探讨经60Co处理的转hGM-CSF基因的HepG2肝癌疫苗的侵袭性和生长活性变化。方法:体外培养三种肝癌细胞(①野生型HepG2肝癌细胞②转染hGM-CSF基因的HepG2肝癌细胞③60Co射线处理的转hGM-CSF基因的HepG2肝癌疫苗)采用MTT方法检测三种细胞在24h、48h、72h的OD值并绘出生长曲线;利用transwell小室进行体外侵袭实验来观察上述三种细胞侵袭性;用RT-PCR技术检测上述三种细胞基质金属蛋白酶2(MMP-2)在mRNA水平上表达的变化;结果:经60Co照射处理的转hGM-CSF基因的HepG2肝癌疫苗组OD值在相同培养时间点较其他两组明显变小且差异有显著性(P<0.05)。三种细胞(上述①②③种细胞)transwell侵袭试验显示:③组穿过人造基底膜的细胞数量明显少于前两组;PT-PCR示:③组细胞的MMP-2的mRNA的表达明显低于①②。结论:经过60Co处理过的转hGM-CSF基因的HepG2肝癌疫苗的侵袭性和生长活性均明显降低。Objective： To investigate the invasion and growth of HepG2 liver cancer vaccines transferred by hGM-CSF gene and which be treated by Co60. Methods： Three hepatoma cells were cultured in vitro （①wild-type HepG2 hepatoma cells②HepG2 hepatoma cells transfected by hGM-CSF gene ③ liver cancer vaccines HepG2 transfected by hGM-CSF gene and irradiated by 60Co）. The OD values of three types of cells cultured at 24h, 48h, 72h was detected by MTT （①② ③ these kinds of cells） and the growth curve was ploted . The three cell invasion was detected by using in vitro transwell chamber invasion assay; The expression of three cell matrix met- alloproteinase 2 （MMP-2） was detected by RT-PCR; Results： 60Co irradiation of hGM-CSF gene transfected liver cancer vaccines HepG2, the growth activity of culture at the same time was significantly lower than that of the other two groups and the difference was significant （P 0.05）. Three kinds of cell （①②③ these kinds of cells） transwell invasion test showed： ③ group the number of cells through artificial basement membrane was significantly less than that of the first two groups; ③ group of cells in the MMP-2 mRNA was significantly lower than that of ①② group. Conclusion： The invasion of HepG2 liver cancer vaccines transferred by hGM-CSF gene and treated by 60Co reduced significantly.

关 键 词：hGM-CSF基因 Transwell体外侵袭 HepG2肝癌疫苗 

分 类 号：Q75[生物学—分子生物学] R735.7[医药卫生—肿瘤]