中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
7期
1524-1526
,共3页
樊清波%马军%董海霞%李印%秦建军%刘红山%秦秉玉
樊清波%馬軍%董海霞%李印%秦建軍%劉紅山%秦秉玉
번청파%마군%동해하%리인%진건군%류홍산%진병옥
白细胞介素-21%细胞因子诱导的杀伤细胞%脱噬作用
白細胞介素-21%細胞因子誘導的殺傷細胞%脫噬作用
백세포개소-21%세포인자유도적살상세포%탈서작용
Interleukin-21%Cytokine-induced killer cells%Apoptosis
目的 观察白细胞介素-21(IL-21)对人细胞因子诱导的杀伤细胞(CIK)体外培养的影响.方法 以常规培养基诱导的CIK细胞为对照组,培养体系中加入IL-21后体外诱导培养10d,观察细胞增殖能力,并检测CIK细胞表型,分析CIK细胞分泌的干扰素(IFN)-γ、肿瘤坏死因子-α(TNF-α)等细胞因子含量,检测CIK细胞对食管癌细胞株EC9706细胞的毒性作用及凋亡.结果 IL-21组细胞数量[(14.58±1.81)×108/L]高于对照组[(7.83±1.18)×108/L,P＜0.05],CD3+细胞比率[(98.48±0.28)％]高于对照组[(95.02±1.18)％,P＜ 0.05],CD3+ CD4+细胞比率[(51.53±1.21)％]高于对照组[(33.36±1.91)％,P＜0.01].IL-21组CD3+ CD8+细胞比率、CD3+ CD56+虽略高于对照组,但差异无统计学意义(P＞0.05).IL-21组培养上清液TNF-α浓度为25.0.μg/L,高于对照组的12.5 μg/L(P ＜0.05),但是两组之间IFN-γ浓度差异无统计学意义(P＞0.05).IL-21组EC9706细胞的凋亡比率为(15.33±0.91)％,高于对照组的(13.25±1.17)％(P＜0.05).结论 IL-21能增加CIK细胞增殖,提高CD3+、CD3+ CD4+阳性表达率,增加细胞因子TNF-α的分泌,促进CIK细胞诱导食管癌细胞凋亡的能力.
目的 觀察白細胞介素-21(IL-21)對人細胞因子誘導的殺傷細胞(CIK)體外培養的影響.方法 以常規培養基誘導的CIK細胞為對照組,培養體繫中加入IL-21後體外誘導培養10d,觀察細胞增殖能力,併檢測CIK細胞錶型,分析CIK細胞分泌的榦擾素(IFN)-γ、腫瘤壞死因子-α(TNF-α)等細胞因子含量,檢測CIK細胞對食管癌細胞株EC9706細胞的毒性作用及凋亡.結果 IL-21組細胞數量[(14.58±1.81)×108/L]高于對照組[(7.83±1.18)×108/L,P＜0.05],CD3+細胞比率[(98.48±0.28)％]高于對照組[(95.02±1.18)％,P＜ 0.05],CD3+ CD4+細胞比率[(51.53±1.21)％]高于對照組[(33.36±1.91)％,P＜0.01].IL-21組CD3+ CD8+細胞比率、CD3+ CD56+雖略高于對照組,但差異無統計學意義(P＞0.05).IL-21組培養上清液TNF-α濃度為25.0.μg/L,高于對照組的12.5 μg/L(P ＜0.05),但是兩組之間IFN-γ濃度差異無統計學意義(P＞0.05).IL-21組EC9706細胞的凋亡比率為(15.33±0.91)％,高于對照組的(13.25±1.17)％(P＜0.05).結論 IL-21能增加CIK細胞增殖,提高CD3+、CD3+ CD4+暘性錶達率,增加細胞因子TNF-α的分泌,促進CIK細胞誘導食管癌細胞凋亡的能力.
목적 관찰백세포개소-21(IL-21)대인세포인자유도적살상세포(CIK)체외배양적영향.방법 이상규배양기유도적CIK세포위대조조,배양체계중가입IL-21후체외유도배양10d,관찰세포증식능력,병검측CIK세포표형,분석CIK세포분비적간우소(IFN)-γ、종류배사인자-α(TNF-α)등세포인자함량,검측CIK세포대식관암세포주EC9706세포적독성작용급조망.결과 IL-21조세포수량[(14.58±1.81)×108/L]고우대조조[(7.83±1.18)×108/L,P＜0.05],CD3+세포비솔[(98.48±0.28)％]고우대조조[(95.02±1.18)％,P＜ 0.05],CD3+ CD4+세포비솔[(51.53±1.21)％]고우대조조[(33.36±1.91)％,P＜0.01].IL-21조CD3+ CD8+세포비솔、CD3+ CD56+수략고우대조조,단차이무통계학의의(P＞0.05).IL-21조배양상청액TNF-α농도위25.0.μg/L,고우대조조적12.5 μg/L(P ＜0.05),단시량조지간IFN-γ농도차이무통계학의의(P＞0.05).IL-21조EC9706세포적조망비솔위(15.33±0.91)％,고우대조조적(13.25±1.17)％(P＜0.05).결론 IL-21능증가CIK세포증식,제고CD3+、CD3+ CD4+양성표체솔,증가세포인자TNF-α적분비,촉진CIK세포유도식관암세포조망적능력.
Objective To analyze the effect of interleukin (IL)-21 on induction of cytokine-induced killer cells (CIK) from human peripheral blood mononuclear cells (PBMCs) in vitro.Methods CIKs were induced from PBMCs of healthy volunteers.The control group was only given conventional medium,and IL-21 was given in IL-21 experimental group.The cells were cultured in an incubator with 5％CO2 at 37 ℃ for 10 days.The proliferation of CIK was analyzed by cell counting trypan blue exclusion test.Fluorescence labeled antihuman CD3,CD4,CD8 and CD56 + antibodies were used for FCM analysis.The cytokines in the culture medium produced by CIK,including interferon (IFN)-γ,tumor necrosis factor (TNF)-α and IL-12,were analyzed by enzyme-linked immunosorbent assay (ELISA).Cytotoxcity on esophageal cancer cells (EC9706) was analyzed by cell counting kit-8 (CCK-8) and apoptosis assays.Results Cell density in IL-21 experimental group was (14.58 ± 1.81) × 108/L,greater than that in control group (7.83 ± 1.18) × 105/L (P ＜ 0.05).In IL-21 experimental group,proportion of CD3 + cells was (98.48 ± 0.28) ％,higher than that in control group [(95.02 ± 1.18) ％] (P ＜ 0.05),and that of CD3+CD4+ was (51.53±1.21)％,higher than in control group [(33.36±1.91)％] (P＜ 0.05).No obvious difference in the proportion of CD3 + CD8 + or CD3 + CD56 + was found between two groups.The concentration of TNF-α in CIK culture medium in IL-21 experimental group was 25.0 μg/L,higher than 12.5 μg/L in control group (P ＜0.05),but there was no significnat difference in IFN-γ concentration.Apoptosis rate was (15.33 ± 0.91) ％ in IL-21 experimental group,higher than (13.25 ± 1.17) ％ in control group (P ＜ 0.05).Conclusion IL-21 can promote the proliferation of CIK.It can increase the proportion of CD3 +,and CD3 + CD4 + in CIK.IL-21 can increase the TNF-o production in CIK,promoting the apoptosis of tumor cells.
