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目的探讨RNA干扰Tax基因表达对人嗜T淋巴细胞病毒Ⅰ型(HTLV-Ⅰ)在T淋巴细胞(TLC)中表达的影响。方法运用RNAi技术特异性干扰Tax基因的表达,流式细胞技术和western blot法鉴定Tax蛋白的表达情况,并通过p27特异性蛋白的检测反映HTLV-Ⅰ在TLC中表达。结果重组质粒psiTax/U6转染T细胞组Tax蛋白的表达(9.84±3.32)%较未处理组(47.45±14.52)%和pSilencer1.0/U6转染组(45.98±10.66)%明显降低;western blot法显示重组质粒psiTax/U6转染TLC后。Tax以及p27蛋白表达较pSilencer1.0/U6转染组明显降低。结论运用RNA技术干扰Tax基因表达可以有效的阻止HTLV-Ⅰ在TLC中的转录。
Objective To investigate the effect of RNA interference (RNAi) Tax gene expression on the expression of human cytotoxic T lymphocyte type Ⅰ (HTLV-Ⅰ) in T lymphocytes (TLC). Methods The RNAi technique was used to specifically interfere with the expression of Tax gene. The expression of Tax protein was identified by flow cytometry and western blot. The expression of HTLV - Ⅰ was detected by TLC in p27 specific protein. Results The expression of Tax protein (9.84 ± 3.32)% in psiTax / U6 transfected T cell group was significantly lower than that in untreated group (47.45 ± 14.52)% and pSilencer1.0 / U6 transfected group (45.98 ± 10.66)% blot showed that the recombinant plasmid psiTax / U6 transfected TLC. Tax and p27 protein expression was significantly lower than pSilencer1.0 / U6 transfection group. Conclusion RNA interference of Tax gene expression can effectively prevent the HTLV-Ⅰ transcription in TLC.