Jurkat T细胞基质金属蛋白酶诱导因子的表达促进单核细胞趋化性

Matrix Metalloproteinase InducerExpressed by Jurkat T Cells Promotes the Chemotaxis of Monocytes

目的通过调控Jurkat T细胞基质金属蛋白酶诱导因子(又称CD147)的表达,观察其对单核细胞趋化性的影响。方法用不同浓度亲环素A(CypA)刺激Jurkat T细胞,采用Western blot检测其CD147蛋白的表达;用CD147特异的小干扰RNA(siRNA)经LipofectamineTM2000转染Jurkat T细胞48 h后,经半定量RT-PCR检测CD147mRNA水平,Western blot检测CD147蛋白水平;用CypA刺激的Jurkat T细胞及CD147基因沉默的Jurkat T细胞(siRNA-Jurkat T)与单核细胞共培养,在趋化实验中观察单核细胞趋化性的变化。结果与对照组相比,0～400μg/L CypA刺激Jurkat T细胞后CD147蛋白的表达上调,呈浓度依赖趋势(P<0.05);而与400μg/L CypA组相比,800μg/L CypA组CD147表达基本无变化(P>0.05)。与阴性对照组相比,CD147特异的siRNA转染Jurkat T细胞后,CD147的mRNA和蛋白表达水平均下降(P<0.05),抑制率分别达76.27%±2.00%和71.67%±4.70%(P<0.05)。CypA预处理的Jurkat T细胞与单核细胞的共培养体系中单核细胞趋化细胞数与空白对照组相比明显增多(196.00±9.88个/视野比109.00±8.22个/视野;P<0.05),siRNA-Jurkat T细胞与单核细胞共培养体系中单核细胞趋化细胞数与空白对照组相比明显下降(44.00±6.98个/视野比109.00±8.22个/视野;P<0.05)。结论 Jur-kat T细胞表达的CD147能够促进共培养体系中单核细胞的趋化性。

Aim To regulate the expression of matrix metalloproteinase inducer （CD147） from Jurkat T cells, and observe its influence on monocyte chemotaxis. Methods Jurkat T cells were stimulated with cyclophilin A （CypA） of different concentrations, and the expression of CD147 protein from Jurkat T cells was determined with Western blot. Jurkat T cells were transfected with CD147-targeting small interfering RNA （siRNA） through the carrier Lipo- fectamineYM2000 for 48 hours. The expression variation of CD147 mRNA and protein were determined by RT-PCR and Western blot respectively. The expression of CD147 from Jurkat T cells was up-regulated with CypA and down-regulated with CD147-targeting siRNA, then these Jurkat T cells and THP-1 monocytes were co-cultured in transwell assay to Count the number of migrated THP-1 ceils. Results Compared with blank control group, the expression of CD147 protein from Jurkat T cells increased along with the concentration gradient of CypA in the range of 0 to 400 μg/L （P 〈 0. 05 ）. Compared with the 400 μg/L CypA group, the expression of CD147 protein was not different With that of the 800 μg/L CypA group （P 〉 0. 05）. Compared with negative control siRNA, CD147 mRNA and protein levels decreased in Jurkat T cells transfected with CD147-targeting siRNA, and the inhibition ratios were 76. 27% ±2. 00% and 71.67% ＋4. 70% （P 〈0. 05）. In the coculture system of Jurkat T cells pretreated with CypA and THP-I cells, the number of migrated THP-1 cells （ 196. 00 ± 9. 88 Cells/Field） significantly increased, compared with blank control group （ 109. 00 ＋8. 22 Cells/Field; P 〈0. 05） ; however, in the coculture system of Jurkat T cells pretreated with siRNA and THP-1 cells, the number of migrated monocytes （44. 00 ＋ 6. 98 Cells/Field） significantly decreased, compared with blank control group （ 109. 00± 8. 22 Cells/Field ; P 〈 O. 05 ）. Conclusions CD147 expressed by Jurkat T cells was able to promote the chemotaxis of monocytes in coculture system.