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目的:分析DLBCL相关抗原特异TCR基因修饰T细胞经肿瘤细胞刺激活化前后TCRζ链基因表达水平的变化。方法:利用SYBRGreenⅠ荧光定量PCR,相对定量检测TCR基因修饰T细胞活化前后TCRζ链基因的表达情况,β2微球蛋白基因(β2M)为内参照,根据相对定量公式:2-△Ct×100%和2-△△Ct,计算TCR基因修饰T细胞活化前后TCRζ链基因的相对表达水平及其差异情况。结果:与未转染T细胞的TCRζ基因mRNA表达量(1.74±0.28)%相比,TCR转基因修饰T细胞TCRζ基因mRNA的表达水平没有发生明显的变化,表达量为(1.78±0.22)%;而经效/靶细胞(TCR基因修饰T细胞/Toledo细胞)混合培养后,活化的TCR基因修饰T细胞的TCRζ基因mRNA表达水平为(11.54±1.98)%,明显高于未刺激培养之前的TCR基因修饰T细胞和未转染T细胞的TCRζ基因的表达水平(P<0.05),分别增长了(6.59±0.80)倍和(6.48±0.36)倍。结论:TCR转基因修饰T细胞受到抗原的刺激而活化,TCRζ链基因的表达相应增加。
OBJECTIVE: To analyze the changes of gene expression of TCRζ chain before and after stimulation of tumor cells by DLBCL-associated TCR gene-modified T cells. Methods: The gene expression of TCRζ chain before and after TCR gene modified T cell activation was detected by SYBR GreenⅠ fluorescence quantitative PCR. Β2 microglobulin gene (β2M) was used as internal reference. According to the relative quantification formula: 2-Δ Ct × 100% and 2- △△ Ct, calculate TCR gene-modified T cell activation TCRζ chain relative expression levels and differences. Results: Compared with untransfected T cells (1.74 ± 0.28)%, the expression of TCRζ mRNA in TCR transgenic T cells did not change significantly (1.78 ± 0.22)%. However, the mRNA expression of TCRζ gene in activated TCR-modified T cells was (11.54 ± 1.98)%, which was significantly higher than that of untreated TCR cells after mixed culture of TCR / T-cell / Toledo cells The expression of TCRζ gene in genetically modified T cells and untransfected T cells increased by (6.59 ± 0.80) times and (6.48 ± 0.36) times, respectively (P <0.05). CONCLUSION: TCR transgenic T cells are activated by antigen stimulation and the expression of TCRζ chain gene is correspondingly increased.