IGF-IR单克隆抗体对Jurkat细胞增殖及凋亡的影响

目的探讨胰岛素样生长因子Ⅰ受体(IGF-IR)在急性T淋巴细胞白血病(T-ALL)细胞系Jurkat的表达、定位,观察IGF-IR单克隆抗体(MAb)对Jurkat细胞增殖、凋亡的影响。方法 (1)采用免疫细胞化学的方法检测IGF-IR在Jurkat细胞的表达、定位。(2)分别用5个不同实验浓度:0.001μg/mL,0.01μg/mL,0.1μ/mL,1μg/mL,10μg/mL的IGF-IR MAb作用Jurkat细胞48h,用CCK-8试剂盒检测细胞的增殖抑制率。(3)选用10μg/mL的IGF-IR MAb作用Jurkat细胞48h,上流式细胞仪测定细胞的凋亡情况。结果 (1)IGF-IR在Jurkat细胞株100%表达,主要为细胞膜、细胞浆混合表达。(2)0.001μg/mL,0.01μg/mL,0.1μg/mL,1μg/mL,10μg/mL的IGF-IR MAb作用Jurkat细胞株48 h,Jurkat细胞的增殖抑制率分别为:(9.67±1.17)%,(14.89±1.06)%,(17.64±0.81)%,(20.15±1.14)%,(24.10±1.11)%,各组间进行两两比较,差异有显著性(P0.05)。(3)10μg/mL的IGF-IR MAb作用Jurkat细胞48h,实验组的诱导细胞凋亡率分别为:早期凋亡率(13.73±1.16)%,晚期凋亡率(20.44±2.47)%,总凋亡率(34.18±3.41)%;对照组的诱导细胞凋亡率分别为:早期凋亡率(6.27±0.67)%,晚期凋亡率(10.18±0.81)%,总凋亡率(16.45±1.38)%。实验组细胞的早期凋亡率,晚期凋亡率,总凋亡率均较对照组高,差异有显著性(P0.05)。结论 (1)Jurkat细胞普遍表达IGF-IR,主要为细胞膜、细胞浆混合表达。(2)IGF-IR MAb可使Jurkat细胞株的增殖受到抑制,且IGF-IR MAb的浓度越高,抑制率越大。(3)IGF-IR MAb可使Jurkat细胞的凋亡增加。

The effects of IGF-IR monoclonal antibody on the proliferation and apoptosis of Jurkat cell line

Objective To explore the expression and cellular location of IGF-Ⅰ R in Jurkat,a T-acute lymphoblastic leukemia (T-ALL) cell line.To investigate the effects of IGF-Ⅰ R monoclonal antibodies (MAb) on the proliferation and apoptosis of Jurkat cell line.Methods (1) The expression level of IGF-Ⅰ R in Jurkat cell line was assessed by immunocytochemistry.(2) Different concentrations of IGF-Ⅰ R MAb of 0.001 μg/mL,0.01 μg/mL,0.1 μg/mL,1 μg/mL and 10 μg/mL were used to treat Jurkat cell lines for 48 hours.CCK-8 assay was used to test the proliferation inhibition rate of Jurkat cell lines.(3)The concentration of 10 μg/mL of IGF-ⅠR MAb was used to treat Jurkat cells for 48 hours.Flow cytometry (FCM) was used to analyzed the apoptosis rates of Jurkat cell lines.Results (1) The positive expression rate of IGF-Ⅰ R was 100％ in Jurkat cell line.The IGF-Ⅰ R as a mixture of both cytoplasm staining and cell membrane was mainly expressed.(2)The inhibition ratios by various concentrations of IGF-Ⅰ R MAb of 0.001 μg/mL,0.01 μg/mL,0.1 μg/mL,1 μg/mL,10 μg/mL for 48 hours were (9.67±1.17)％,(14.89 ±1.06)％,(17.64±0.81)％,(20.15 ±1.14)％,and (24.10 ± 1.11)％,respectively.Significant statistics difference was found in multiple comparison (P ＜0.05).(3) In the trial group,the early apoptosis rate of Jurkat cells was (13.73 ± 1.16)％,the late apoptosis rate of Jurkat cells was (20.44 ± 2.47) ％,the total apoptosis rate of Jurkat cells was (34.18 ± 3.41)％.While in the control group,the early apoptosis rate of Jurkat cells was (6.27 ± 0.67)％,the late apoptosis rate of Jurkat cells was (10.18 ± 0.81) ％,the total apoptosis rate of Jurkat cells was (16.45 ± 1.38)％.The differences between the trial group and control group had statistical significance (P ＜ 0.05).Conclusions (1) The IGF-Ⅰ R was generally expressed in Jurkat cell line and the IGF-Ⅰ R as a mixture of both cytoplasm staining and cell membrane was mainly expressed.(2) IGF-Ⅰ R MAb produced dose-dependent inhibition of Jurkat cell growth,higher concentration has better proliferation inhibition rate.(3) IGF-Ⅰ R MAb could promote the apoptosis of Jurkat cell line.