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目的应用小干扰RNA(siRNA)技术转染Jurkat细胞,探讨CD147基因沉默后对Jurkat细胞增殖、CD147的表达以及侵袭力的影响。方法实验分3组:空白对照组即Jurkat细胞组,阴性对照组即转染阴性质粒的pGE-Ⅰ-CD147(-)-shRNA组,实验组即转染重组质粒的pGE-Ⅰ-CD147(+)-shRNA。RT-PCR检测转染前后细胞CD147 mRNA表达水平;流式细胞仪检测转染前后细胞周期变化;基质黏附试验检测转染后细胞基质黏附能力的变化。结果pGE-I-CD147(+)-shRNA的Jurkat细胞CD147 mRNA表达较空白对照组明显下降(CD147/β-actin灰度比:0.35±0.03vs0.92±0.13P<0.05);pGE-I-CD147(+)-shRNA的Jurkat细胞G0+G1期细胞比例较空白对照组明显增多[(89.85±7.85)%vs(57.55±5.64)%P<0.05];pGE-I-CD147(+)-shRNA的Jurkat细胞黏附率较空白对照组明显增多[(21.74±7.65)%vs(47.12±4.55)%P<0.05]。结论通过siRNA技术沉默CD147基因表达可抑制Jurkat细胞增殖,降低CD表达及细胞的侵袭力,从而为以CD为靶点的基因治疗非霍奇金淋巴瘤提供实验依据。
Objective To investigate the effect of CD147 gene silencing on Jurkat cell proliferation, CD147 expression and invasiveness in Jurkat cells transfected with small interfering RNA (siRNA). Methods The experiment was divided into three groups: the blank control group was Jurkat cell group, the negative control group was transfected with negative plasmid pGE-Ⅰ-CD147 (-) - shRNA group, the experimental group transfected with recombinant plasmid pGE-Ⅰ-CD147 (+ ) -shRNA. The expression of CD147 mRNA was detected by RT-PCR before and after transfection. The changes of cell cycle before and after transfection were detected by flow cytometry. The changes of matrix adhesion capacity after transfection were detected by matrix adhesion assay. Results The expression of CD147 mRNA in Jurkat cells of pGE-I-CD147 (+) - shRNA was significantly decreased compared with the blank control group (gray scale ratio of CD147 / β-actin: 0.35 ± 0.03 vs 0.92 ± 0.13 P < The percentage of G0 + G1 phase cells in Jurkat cells with CD147 (+) - shRNA was significantly increased compared with that in the blank control group [(89.85 ± 7.85)% vs (57.55 ± 5.64)% P <0.05] Jurkat cell adhesion rate was significantly increased compared with the control group [(21.74 ± 7.65)% vs (47.12 ± 4.55)% P <0.05]. Conclusion Silencing the expression of CD147 gene by siRNA can inhibit the proliferation of Jurkat cells and decrease the expression of CD and the invasiveness of cells, so as to provide an experimental basis for CD gene therapy for non-Hodgkin’s lymphoma.