针对半乳糖凝集素-3的siRNA抑制CD133~+肺腺癌细胞体外诱导CD8~+T细胞凋亡

目的 观察针对半乳糖凝集素(galectin)-3的特异性siRNA对CD133~+肺腺癌细胞功能的影响.方法 实时荧光定量PCR技术(FQRT-PCR)和Western blot检测siRNA对galectin-3的干扰效率,噻唑蓝(MTT)比色法检测siRNA对CD133~+肺腺癌细胞增殖的影响,Annexin V和PI双染检测转染siRNA的CD133~+肺腺癌细胞上清诱导CD8+T细胞凋亡能力.结果 针对galeetin-3的siRNA平均干扰效率为80.3%,明显降低CD133~+细胞中galectin-3的表达,并抑制CD133~+细胞增殖,转染96 h后细胞活率为未转染组的(75.0±3.5)%,凋亡检测结果表明转染siRNA的CD133~+肺腺癌细胞上清诱导CD8~+T细胞凋亡率为8.2%,与未转染组凋亡率18.6%比较诱导凋亡的能力明显下降,差异有统计学意义(P＜0.05).结论 针对galectin-3的siRNA高效抑制galectin-3的表达,降低细胞的增殖能力及诱导CD8~+T细胞凋亡的能力,具有潜在的临床应用价值.

Inhibitory effects of specie siRNA targeting-3 on CB8~+ T cell apoptosis induced by CD133~+pulmonary adenocarcinoma cells

Objective To explore the inhibitory effects of specific siRNA targeting galectin-3 on the function of CD133~+ lung adenocaxeinoma cells.Methods The effect of galeetin-3 siRNA was detected by fluorescent quantitative reverse transcription-polymerage chain reaction(FQRT-PCR)and Western blotring,and the inhibitory effect of galectin-3 on the growth of CD133~+lung adenocarcinoma cells was detected by MTY assay.Annexin V and PI test wag used to investigate whether supernatants of CD133~+lung adenoearcinoma ceHs transfeeted with siRNA or siRNAmut could mediate apoptosis of CD8~+T cells in vitro.Results The results of F9RT-PCR revealed that siRNA could interfere with galeetin-3 with the interferenee efficiency being 80.3%.The Western blotting results showed that siRNA could efliciently depressed the galectin-3 protein expression and the mutant of siRNA had no effect to interfere with galeetin-3.The MTT assay demonstrated that the viability rate of CD133~+ ceils treated with siRNA was decreased with with time after transfection.At the 96th h after transfection,the viability rate of CD133~+ cells treated with siRNA[(75.0±3.5)%]was signifieanfly lower than that of CD133~+cells treated with sirNA mutant [(95.0±1.2)%].The apoptosis rate of CD8+ T cells indueed by the supernatants of CD133~+ cells at the 48th h after transfection with siRNA was 8.2%.which was lower than that of CD133~+ cells untreated (18.6%)(P＜0.05).Conclusion Down-regulation of galectin-3 by siRNA could efficiendy reduce the proliferative capacity of CD133~+ cells in vitro as well as induce CD8~+ T cell apoptosis.These findings indicate Galectin-3 may play an important role during oncogenesis,implying a potential therapeutic value of siRNA targeting galectin-3 in clinical practice.