目的 研究高迁移率蛋白A1(HMGA1)在人结肠癌干细胞维持中的表达与功能.方法 利用实时定量反转录聚合酶链反应(RT-qPCR)和Western blot检测HMGA1在结肠癌干细胞(CTSC)、普通结肠癌细胞株HCT116、HT29和对照结肠黏膜(NM)中的表达;利用小干扰RNA(siRNA)抑制HMGA1的表达,进行干细胞增殖、克隆形成能力分析,研究HMGA1在结肠癌干细胞自更新能力中的作用.结果 HMGA1 mRNA在NM组织中相对表达量分别为1.00±0.09,而在HCT1 16、HT29细胞中HMGA1 mRNA相对表达量为1.56 ±0.13、1.62±0.10,显著高于NM组(t=5.949,P=0.005;t =8.124,P=0.001).在CTSC中,HMGA1 mRNA表达量为2.37±0.15,明显高于NM组(t=12.330,P=0.001);HMGA1沉默的CTSC细胞增殖显著削弱(3.60±0.14比2.71±0.30,t=4.649,P=0.010).HMGA1沉默的CTSC细胞克隆的个数和直径分别为55 710±11 007(t=5.957,P=0.004)和(0.50±0.04) mm(t=3.499,P=0.025),均显著下降,差异具有统计学意义.结论 靶向HMGA1的siRNA可抑制结肠癌干细胞的增殖和克隆形成,提示HMGA1参与促进结肠癌干细胞的自更新能力.
Objective To investigate the expression and role of high-mobility group A1 (HMGA1) in maintaining of colon tumour stem cell (CTSC).Methods Real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and Westem blotting were applied to detect the expression of HMGA1 in CTSCs,regular colon cancer cells (HCT116 and HT29) and nornal colonic mucosa (NM).Synthetic small interfering RNAs (siRNAs) were used to suppress the intrinsic HMGA1 expression.CTSC self-renewal was assessed by methyl thiazol tetrazolium (MTT) proliferation assay and sphere-forming efficiency.Results HMGA1 mRNA relative expression were 1.00 ±0.09,1.56 ±0.13 and 1.62 ±0.10 for NM,HCT116 and HT29 respectively with statistical difference (t =5.949,P =0.005;t =8.124,P =0.001);HMGA1 mRNA expression was 2.37 ±0.15 which was statistical higher than NM group (t =12.330,P =0.001);The cell proliferation ability was significant decreased by inhibiting HMGA1 (3.60 ± 0.14 vs.2.71 ± 0.30,t =4.649,P =0.010);The number and diameter of HMGA1 silenced CTSC cell clones were 55 710 ± 11 007 (t =5.957,P =0.004) and (0.50 ± 0.04) mm(t =3.499,P =0.025) decreased significantly with statistically significant difference.Conclusion HMGA1 interfering could significantly inhibit cell proliferation and sphere formation,which suggest its critical role involved in self-renewal of CTSCs.