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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 invasiveness and growth activity of HepG2 hepatocarcinoma vaccine transfected with 60Co-treated hGM-CSF gene. Methods: Three kinds of hepatoma cells were cultured in vitro (① wild-type HepG2 hepatocellular carcinoma cells ② HepG2 hepatoma cells transfected with hGM-CSF gene ③ HepG2 hepatocarcinoma vaccine transfected with hGM-CSF gene by 60Co radiotherapy) 48h and 72h respectively. The invasiveness of the three kinds of cells was observed by using transwell chamber in vitro. The expression of matrix metalloproteinase 2 (MMP-2) at the mRNA level .Results: The OD values of HepG2 hepatocarcinoma cells transfected with 60Co irradiation at the same culture time point were significantly smaller than those in the other two groups (P <0.05). Transwell invasion assay showed that: (3) The number of cells crossing the artificial basement membrane was significantly less than that of the first two groups; PT-PCR showed: (3) The mRNA expression of MMP-2 in the group of cells was significantly lower than ① ②. Conclusion: The invasive and growth activities of HepG2 hepatocarcinoma cells transfected with hGM-CSF gene after 60Co treatment were significantly decreased.