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

Inhibitory effects of specific siRNA targeting-3 on CD8＋ T cell apoptosis induced by CD133＋ pulmonary adenocarcinoma cells

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Westernblot检测siRNA对galectin．3的干扰效率，噻唑蓝（MTr）比色法检测siRNA对CD133＋肺腺癌细胞增殖的影响，AnnexinV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galectin-3的siRNA平均干扰效率为80．3％，明显降低CD133＋细胞中galeetin-3的表达，并抑制CD133＋细胞增殖，转染96h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。

Objective To explore the inhibitory effects of specific siRNA targeting galectin-3 on the function of CD133＋ lung adenocarcinoma cells. Methods The effect of galectin-3 siRNA was detected by fluorescent quantitative reverse transcription-polymerase chain reaction （FQRT-PCR） and Western blotting, and the inhibitory effect of galectin-3 on the growth of CD133＋ lung adenocarcinoma cells was detected by MTT assay. Annexin V and PI test was used to investigate whether supernatants of CD133 ＋ lung adenocarcinoma ceils transfected with siRNA or siRNAmut could mediate apoptosis of CD8 ＋ T ceils in vitro. Results The results of FQRT-PCR revealed that siRNA could interfere with galectin-3 with the interfer- ence efficiency being 80.3%. The Western blotting results showed that siRNA could efficiently depressed the galeetin-3 protein expression and the mutant of siRNA had no effect to interfere with galectin-3. The MTT assay demonstrated that the viability rate of CD133＋ cells treated with siRNA was decreased with with time after transfection. At the 96th h after transfection, the viability rate of CD133＋ cells treated with siR- NA [ （75.0 ±3.5）% ] was significantly lower than that of CD133＋ cells treated with siRNA mutant [ （95.0 ±1.2）% ]. The apoptosis rate of CD8 ＋ T cells induced 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 efficiently reduce the proliferative capacity of CD133 ＋ cells in vitro as well as induce CD8＋ T cell apoptosis. These findings indicate Galeetin-3 may play an important role during oncogenesis, implying a potential therapeutic value of siRNA targeting galeetin-3 in clinical practice.