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目的:筛选针对乳腺癌有效的hTERT siRNA,为应用RNAi技术在乳腺癌中敲除hTERT基因研究提供实验基础。方法:采用化学合成法合成3段针对hTERT的siRNA、应用脂质体转染试剂分别转染人乳腺癌MCF-7细胞、实时荧光定量RT-PCR技术检测hTERT mRNA表达水平以确定干扰效果。结果:3段hTERT siRNA中有2段siRNA可有效降低hTERT mR-NA表达。结论:利用化学合成法合成siRNA,应用实时荧光定量PCR技术检测目的基因干扰效果,可快速、高效筛选特异性抑制基因表达的siRNA。
Objective: To screen effective hTERT siRNA against breast cancer and provide experimental basis for the study of hTERT gene knockdown in breast cancer using RNAi technique. Methods: Three sections of siRNA against hTERT were synthesized by chemical synthesis. Human breast cancer cell line MCF-7 was transfected with lipofectamine. The expression of hTERT mRNA was detected by real-time fluorescence quantitative RT-PCR to determine the interference effect. Results: Two hTERT siRNAs in 3 hTERT siRNAs could effectively reduce the expression of hTERT mR-NA. CONCLUSIONS: siRNA is synthesized by chemical synthesis and real-time fluorescent quantitative PCR is used to detect the interference effect of the target gene. The siRNAs that specifically inhibit the gene expression can be rapidly and efficiently screened.