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探讨靶向STAT3的小分子干扰RNA(STAT3-siRNA)对多柔比星(doxorubicin,DOX)的抗肿瘤活性的影响。通过RT-PCR及Western blotting检测STAT3在HepG2、HeLa和K562/DOX细胞中的表达水平以及小分子干扰RNA对STAT3表达的影响。采用MTT法和台盼蓝染色法,检测STAT3-siRNA对肿瘤细胞的抑制作用及其对DOX抗肿瘤活性的影响。应用高内涵活细胞成像系统检测STAT3-siRNA对细胞内DOX蓄积量以及细胞凋亡的影响。结果显示,STAT3在HepG2、HeLa和K562/DOX细胞中均有高表达,STAT3-siRNA明显下调STAT3mRNA及STAT3蛋白的表达。STAT3-siRNA可抑制HepG2、HeLa和K562/DOX细胞的生长,STAT3-siRNA与DOX联合应用后对3种细胞的IC50分别降低了3.13倍、5.22倍和1.74倍(与DOX单独应用比较)。STAT3-siRNA使HepG2、HeLa及K562/DOX细胞内DOX的蓄积量分别增加16.8%、12.87%和25.67%,同时明显促进了DOX诱导的细胞凋亡。结果表明,STAT3-siRNA可明显增强DOX的抗肿瘤活性。
To investigate the effect of small interfering RNA (STAT3-siRNA) targeting STAT3 on the antitumor activity of doxorubicin (DOX). The expression of STAT3 in HepG2, HeLa and K562 / DOX cells and the effect of small interfering RNA on STAT3 expression were detected by RT-PCR and Western blotting. The inhibitory effect of STAT3-siRNA on tumor cells and its effect on the antitumor activity of DOX were detected by MTT and trypan blue staining. The high content of living cell imaging system was used to detect the effect of STAT3-siRNA on DOX accumulation and cell apoptosis. The results showed that STAT3 was highly expressed in HepG2, HeLa and K562 / DOX cells, STAT3-siRNA significantly down-regulated STAT3 mRNA and STAT3 protein expression. STAT3-siRNA inhibited the growth of HepG2, HeLa and K562 / DOX cells. The IC50 of STAT3-siRNA and DOX decreased by 3.13, 5.22 and 1.74 fold compared with DOX alone. The accumulation of DOX in HepG2, HeLa and K562 / DOX cells was increased by 16.8%, 12.87% and 25.67% respectively by STAT3-siRNA, and DOX-induced apoptosis was also significantly promoted. The results show that, STAT3-siRNA can significantly enhance the anti-tumor activity of DOX.