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目的探讨外源性IL-18基因表达对C6胶质瘤细胞侵袭力的影响。方法无菌条件下在Boyden小室下室中加入NIH3T3细胞培养液上清200μl,在上室中分别加入C6/IL-18、C6/pLX-SN、C6细胞悬液,用含Matrigel Matrix的聚碳酸酯膜将上、下室隔开。培养24h后,取出聚碳酸酯膜,甲醇固定,苏木精染色,在高倍光镜下计数穿过每张膜的细胞数。对数生长期C6/IL-1、C6/pLXSN和C6细胞,接种于24孔培养板,37℃、5%CO2培养箱培养12d。培养结束时,甲醇固定15min,姬姆萨染液染色30min。在50倍光镜下观察集落形成,计算集落形成率。结果 C6/IL-18细胞、C6/pLXSN细胞及C6细胞平均每个视野穿过聚碳酸酯膜的细胞数分别为(11.25±1.20)个、(16.50±1.77)个和(17.05±1.47)个细胞。C6/IL-18细胞与C6细胞和C6/pLXSN细胞比较差异有统计学意义(P<0.05),C6/pLXSN细胞与C6细胞比较穿过膜的细胞数差异无统计学意义(P>0.05)。C6/IL-18细胞、C6/pLXSN细胞及C6细胞克隆形成率分别为(5.44±0.63)%、(8.35±0.65)%和(8.48±0.76)%。C6/IL-18细胞的克隆形成率降低,与C6细胞和C6/pLXSN细胞比较差异有统计学意义(P<0.05),C6细胞和C6/pLXSN细胞比较差异无统计学意义(P>0.05)。结论外源性IL-18基因表达可降低C6胶质瘤细胞侵袭力和克隆形成。
Objective To investigate the effect of exogenous IL-18 gene on invasiveness of C6 glioma cells. Methods A total of 200 μl of NIH3T3 cell culture supernatant was added to the lower chamber of Boyden chamber under aseptic conditions. C6 / IL-18, C6 / pLX-SN and C6 cell suspension were added to the upper chamber, Ester film will be separated from the lower chamber. After culturing for 24 hours, the polycarbonate membrane was removed, fixed in methanol and stained with hematoxylin and the number of cells that passed through each membrane was counted under a high power light microscope. Logarithmic growth phase C6 / IL-1, C6 / pLXSN and C6 cells were seeded in 24-well culture plates, 37 ° C, 5% CO2 incubator for 12 days. At the end of the culture, the methanol was fixed for 15 min and the Giemsa stain was stained for 30 min. Colony formation was observed under a 50-fold light microscope to calculate the rate of colony formation. Results The average number of C6 / IL-18 cells, C6 / pLXSN cells and C6 cells passing through the polycarbonate membrane was (11.25 ± 1.20), (16.50 ± 1.77) and (17.05 ± 1.47) cell. C6 / IL-18 cells compared with C6 cells and C6 / pLXSN cells (P <0.05), C6 / pLXSN cells and C6 cells compared with the number of cells through the membrane was no significant difference (P> 0.05) . The formation rates of C6 / IL-18 cells, C6 / pLXSN cells and C6 cells were (5.44 ± 0.63)%, (8.35 ± 0.65)% and (8.48 ± 0.76)%, respectively. (P <0.05). There was no significant difference between C6 cells and C6 / pLXSN cells (P> 0.05), but there was no significant difference between C6 cells and C6 / . Conclusion Exogenous IL-18 gene expression can reduce invasiveness and clonogenicity of C6 glioma cells.