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目的:研究rmhTNF-α联合抗IL-6抗体对胃癌SGC-7901细胞的杀伤作用。方法:(1)将已培养的SGC-7901细胞株分为a、b、c、d、e组,a、b、c、d组分别加入浓度为10 ng/ml、20 ng/ml、50 ng/ml、100 ng/ml的rmhTNF-α,e组加入等量PBS。ELISA检测不同时间点SGC7901细胞上清液中IL-6的分泌量,MTT法检测细胞培养24 h、32 h、40 h、48 h的凋亡情况。(2)将SGC-7901细胞分为f、g、h、i组,f组加入rmhTNF-α,浓度分别为10 ng/ml、20 ng/ml、50 ng/ml、100 ng/ml;g组加入不同浓度的rmhTNF-α与浓度为100 ng/ml的IL-6,h、i组加入不同浓度的rmhTNF-α与浓度分别为1:62.5和1:125的抗IL-6抗体。MTT法测细胞培养48 h的凋亡情况。荧光显微镜观察细胞凋亡情况。结果:(1)IL-6分泌五组差别具有统计学意义(P<0.05),a、b、c、d组IL-6浓度均高于e组(P<0.05),a、b、c组相比各时间点IL-6浓度逐渐升高(P<0.05),c、d组差别无统计学意义。(2)细胞存活率a、b、c、d组与e组差别具有统计学意义(P<0.05),a、b、c、d组内比较差别有统计学意义(P<0.05),显示杀伤作用40 h后减弱,c、d组差别无统计学意义。(3)细胞存活率f、g组比较差别具有统计学意义(P<0.05),h、i组与f组差别有意义(P<0.05),h、i组比较,i组细胞存活率较低。结论:rmhTNF-α可以促进SGC-7901细胞分泌IL-6;抗IL-6抗体可以增强rmhTNF-α对SGC-7901细胞的杀伤作用。
Objective: To study the killing effect of rmhTNF-α combined with anti-IL-6 antibody on gastric cancer SGC-7901 cells. Methods: (1) The cultured SGC-7901 cell line was divided into groups a, b, c, d and e, and groups a, b, c and d were respectively treated with 10 ng / ml, 20 ng / ng / ml, 100 ng / ml of rmhTNF-α, e group added an equal amount of PBS. The levels of IL-6 in supernatant of SGC7901 cells were detected by ELISA. The apoptosis of SGC7901 cells was detected by MTT assay at 24 h, 32 h, 40 h, 48 h. (2) SGC-7901 cells were divided into groups f, g, h and i, group f was added with rmhTNF-α at concentration of 10 ng / ml, 20 ng / ml, 50 ng / ml and 100 ng / Groups were given different concentrations of rmhTNF-α and IL-6 at a concentration of 100 ng / ml. Group i was given anti-IL-6 antibody with different concentration of rmhTNF-α and concentrations of 1: 62.5 and 1: 125 respectively. The apoptosis of cells cultured for 48 h was measured by MTT assay. Fluorescence microscopy observation of apoptosis. The levels of IL-6 in a, b, c and d groups were significantly higher than those in e group (P <0.05), a, b and c Compared with each time point, the concentration of IL-6 gradually increased (P <0.05), while there was no significant difference between c and d. (2) The cell survival rates in a, b, c and d groups were significantly different from those in e group (P <0.05), but there was significant difference in a, b, c and d groups The killing effect was weakened after 40 h, but there was no significant difference between c and d groups. (3) There was significant difference between group f and group g (P <0.05), but there was significant difference between group h and group f (P <0.05). The survival rate of group i was higher than group i low. Conclusion: rmhTNF-α can promote the secretion of IL-6 by SGC-7901 cells. Anti-IL-6 antibody can enhance the killing effect of rmhTNF-α on SGC-7901 cells.