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目的研究小干扰RNA抑制CD147基因对Jurkat T淋巴细胞集落形成的影响。方法将CD147特异的siRNA,经LipofectamineTM2000转染Jurkat细胞48 h后,用半定量RT-PCR检测CD147 mRNA水平的变化,Western blot和流式细胞术(FCM)分别检测总蛋白和细胞表面蛋白水平的变化。通过荧光倒置显微镜比较特异性抑制CD147表达后Jurkat细胞集落形成的变化。结果与对照组相比siRNA干扰组细胞CD147 mRNA和蛋白水平的表达均降低,抑制率分别为(38.57±1.55)%和(47.4±1.47)%,细胞表面CD147的平均荧光强度(MFI)下降(50.5±4.7)%(P<0.05)。随着CD147被抑制,siRNA干扰组细胞集落形成明显少于对照组(P<0.05)。结论通过siRNA能抑制Jurkat细胞CD147的表达,随着CD147表达下降细胞集落的形成减少,提示CD147可能在炎症细胞粘附聚集中起到一定作用。
Objective To investigate the effect of small interfering RNA (CDNA) on CD147 Jurkat T lymphocyte colony formation. Methods CD147-specific siRNA was transfected into Jurkat cells with LipofectamineTM2000 for 48 h. The level of CD147 mRNA was detected by semi-quantitative RT-PCR. The protein and cell surface protein levels were detected by Western blot and flow cytometry (FCM) Variety. Fluorescent inverted microscope was used to compare the changes of Jurkat cell colony formation that specifically inhibited CD147 expression. Results Compared with the control group, the expression of CD147 mRNA and protein in the siRNA - siRNA group decreased (38.57 ± 1.55)% and (47.4 ± 1.47)%, respectively. The mean fluorescence intensity (MFI) of CD147 on the cell surface decreased 50.5 ± 4.7)% (P <0.05). With the inhibition of CD147, colony formation of siRNA interference group was significantly less than that of control group (P <0.05). Conclusion siRNA can inhibit the expression of CD147 in Jurkat cells. With the decrease of CD147 expression, the formation of colony is reduced, suggesting that CD147 may play a role in the adhesion and aggregation of inflammatory cells.