摘要
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(P<0.05)。(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(P<0.05)。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。
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 p,g/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.
作者
郑丽娟
张文林
ZHENG Lijuan ZHANG Wenlin.(Department of Pediatrics, Zhengzhou Children＇s Hospital, Zhengzhou 450000, Chin)
出处
《中国小儿血液与肿瘤杂志》
CAS
2017年第4期194-199,共6页
Journal of China Pediatric Blood and Cancer
基金
河南省教育厅科技攻关计划项目(No.200510472005)
作者简介
通讯作者：张文林，Email：wenlin331800@sina．com
