摘要
目的:观察抑制hsa-miR-204表达对人急性淋巴细胞白血病细胞(MOLT-4)生长与凋亡的影响。方法:设计合成hsa-miR-204的反义核苷酸序列(AMO-miR-204)并用脂质体转染法将其转染MOLT-4细胞。Q-PCR检测hsa-miR-204的表达量。噻唑蓝(MTT)试验和平板克隆形成实验检测细胞增殖能力;流式细胞术检测细胞早期凋亡率。Transwell检测细胞侵袭能力。结果:AMO-miR-204可抑制hsa-miR-204的表达,以反义核酸浓度为0.6μmol/L时,下调最为明显(P<0.05)。MTT实验示AMO-miR-204的最佳转染抑制浓度为0.6μmol/L。AMO6组细胞平板克隆形成能力明显减弱[(29±8)%],差异有统计学意义(P<0.05)。流式细胞术检测AMO6组细胞凋亡达(7.47%)。Transwell细胞侵袭能力实验示AMO6组细胞侵袭能力较其余两组减弱。结论:AMO-miR-204能有效抑制MOLT-4细胞的增殖,并促进其凋亡。hsamiR-204有可能作为急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)基因治疗的靶基因,同时也为深入揭示ALL的发生和发展机制提供了实验依据。
Objective ：To observe the effect of hsa- miR -204 inhibitor on human acute lymphoblastic leukemia cell growth. Methods ： Designed and synthetized the inhibitor of hsa - miR - 204, AMO - miR - 204, and transfected the MOLT -4 cell by liposome. Real -time quantitative PCR（ Q -PCR） was used to detect hsa -miR- 204 expres- sion after transfection. The MOLT- 4 cell proliferation was evaluated by Methyl thiazol terazolium（MTT） assay and soft agar assay. The apoptosis rate was tested by flow cytometry, and the invasive ability was measured by Transwell. Results：The antisense oligonucleotide AMO -miR- 204 effectively down- regulated the hsa- miR- 204 expression in MOLT -4 cells. Especially when the concentration of AMO - miR -204 was 0.6μmol/L, the decrease of hsa - miR- 204 was the most obvious,and the MTT？ test demonstrated that this concentration was the best transfection inhi- bition concentration to the apoptosis of MOLT - 4 cells. The tablet colony - forming ability of cells from group AMO6 was significantly suppressed [ （ 29 ± 8 ） % ], and the differences were statistically significant （ P 〈 0.05 ）. The Transwell test showed that the cell invative ability was suppressed in group AMO6. Conclusion ： AMO - miR - 204 could inhibit the proliferative ability and induce apoptosis of MOLT - 4 cells, prompting that hsa - miR - 204 can possibly be the target gene of acute lymphoblastic leukemia（ ALL ）gene treatment,and providing the experimental evidence to uncov- er the occurrence and development mechanism of ALL at the same time.
出处
《现代肿瘤医学》
CAS
2013年第12期2639-2643,共5页
Journal of Modern Oncology
基金
广东省中医药管理局项目(编号:20121018)
