rmhTNF-α联合抗IL-6抗体对胃癌SGC-7901细胞杀伤作用的研究

Studies on the Cytotoxicity of RmhTNF-α Combined with Anti-IL-6 Antibody to SGC-7901 Cell Line

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 investigate the cytotoxicity of rmhTNF-α combined with anti-IL-6 antibody to SGC-7901 cell line.Methods：（1）SGC-7901 cell line was divided into groups a,b,c,d,e,rmhTNF-α（10 ng/ml,20 ng/ml,50 ng/ml,100 ng/ml）was added into the a,b,c,d groups respectively,while the same amount of PBS was put into the e group.The secretion of IL-6 of SGC-7901 cell line at 4 h,8 h,12 h,16 h,20 h,24 h was detected by ELISA.MTT assay was used to definite cell apoptosis at 24 h,32 h,40 h,48 h.（2）SGC-7901 cell line was divided into groups f,g,h,i,and rmhTNF-α was added into the f group,with concentrations 10 ng/ml,20 ng/ml,50 ng/ml,100 ng/ml;RmhTNF-α and 100 ng/ml of IL-6 were put into the g group;RmhTNF-α was combined with anti-IL-6 antibody into the h and i group,and the concentrations of anti-IL-6 antibody were 1：62.5 and 1：125.The cell apoptosis was detected by MTT assay at 48 h.Cell apoptosis was observed with a fluorescence microscopy.Results：（1）The secretion of IL-6 of the e group was lower than o-ther groups（P0.05）,and there was no significant difference between Group c and Group d.（2）The cell viability difference between the e group and other groups was significant（P0.05）,and the difference in the a,b,c,d group was significant,which showed the cytotoxicity decreased after 40 h,and we found no difference between the c and d group.（3）The cell viability difference between the f and g group was significant（P0.05）,and the difference among groups h,i and f was significant（P0.05）.Compared the h and i group,cell viability of the h group was lower.Conclusion： RmhTNF-α could promote SGC-7901 cell secret IL-6;Anti-IL-6 antibody strength cytotoxicity of rmhTNF-α to SGC-7901 cell line.