摘要
目的体外设计、合成及筛选高效沉默同源盒基因A7（HOXA7）表达的小干扰RNA（siRNA），并研究沉默HOXA7对人类肺腺癌细胞株LETP-a-2增殖、凋亡的影响。方法设计、合成三对针对HOXA7基因的siRNA（siRNAl、siRNA2、siRNA3）和一对阴性对照（siRNAnc），分别用阳离子脂质体介导转染LETPIa-2细胞。实验分6组：细胞对照组、脂质体对照组、siRNAnc组、siRNAl组、siRNA2组、siRNA3组，反转录聚合酶链反应（RT．PCR）和Westernblot分别检测各组HOXA7mRNA和蛋白表达情况，四甲基偶氮唑蓝（MTT）比色法和流式细胞术分别检测沉默HOXA7后LETP—a-2细胞增殖、凋亡情况。结果siRNA成功转染LETP-a-2细胞，三对siRNA均能有效沉默HOXA7表达，其中以siRNA2沉默HOXA7的效果最佳，在mRNA和蛋白水平沉默率分别为（57．344±4．743）％和（52．219±0．550）％；三对siRNA沉默HOXA7后均能抑制LETP-a-2细胞增殖并促进其凋亡，其中以siRNA2组效果最明显，48h细胞增殖抑制率可达（48．144±4．992）％，凋亡率达（26．613±0．612）％。结论筛选出高效沉默HOXA7的siRNA2能有效抑制LETP—a-2细胞增殖并促进其凋亡，提示HOXA7可能成为肿瘤基因治疗的新靶点。
Objective To design and screen small interefere RNA （siRNA） targeting of HOXA7, and to investigate the effect of the siRNA on human lung cancer LETP-a-2 cells proliferation and apoptosis in vitro. Methods Three pairs of siRNA targeting of HOXA7 and one pair of siRNA for negative control were transfected respectively into LETP-a-2 cells through cationic liposome. The mRNA and proteion expression levels of HOXA7 were observed by reverse transcription polymerase chain reaction （RT-PCR） and Western blot analysis. The effect of HOXA7 siRNA on growth and apoptosis of LETP-a-2 cells were measured by MTr and flow cytometry. Results All the three pairs of siRNA could inhibit HOXA7 expression effectively, among which siRNA2 got the best effects, the silence rates were （57.344±4.743） % on mRNA level and （52.219±0.550） % on protein leval. The proliferation was inhibited and the apoptosis was promoted by the siRNA targeting HOXA7 in LETP-a-2 cells, among which siRNA2 got the favourite results, the inhibitory rate was （48.144±4.992） % and the apoptosis rate was （26.613±0.612） %. Conclusion The siRNA2 targeting of HOXA7 enrolls in promoting apoptosis and inhibiting grows of LETP-a-2 cells, indicating that manipulation of HOXA7 expression may be a potential therapeutic strategy for cancer.
出处
《肿瘤研究与临床》
CAS
2013年第9期581-584,587,共5页
Cancer Research and Clinic
基金
山东省科学技术发展计划（2010GSF10264）
