RNA干扰沉默HOXA2基因表达对肝癌细胞株行为的影响  

Effect of homeobox A2 gene RNA interference on HCC cell behavior

作　　者：胡晓珺[1] 王一成 欧伶[1] 

机构地区：[1]华东理工大学生物工程学院,上海200237 [2]上海南方模式生物研究中心,上海201203

出　　处：《安徽医科大学学报》2014年第2期154-159,共6页Acta Universitatis Medicinalis Anhui

基　　金：上海市自然科学基金(编号:07ZR14082)

摘　　要：目的研究同源异型盒A2(HOXA2)基因对肝癌细胞生长、细胞周期及凋亡的影响,探讨其作为潜在肝癌治疗靶点的可行性。方法采取实时定量PCR和逆转录PCR检测HOXA2基因的表达情况;siRNA干扰HOXA2基因在肝癌细胞株PLC/PRF/5、MHCC-97L中表达,通过CCK-8评价肝细胞肝癌细胞株增殖情况,采用软琼脂克隆形成试验评价锚泊非依赖生长情况,流式细胞术分析细胞周期及细胞凋亡。结果实时定量PCR和逆转录PCR检测结果显示HOXA2基因在肝癌组织中表达上调(P<0.05)。针对HOXA2基因设计合成siRNAs能够特异性沉默HOXA2基因在肝癌细胞株PLC/PRF/5、MHCC-97L中的表达,在干扰了HOXA2基因表达后肝癌细胞株的软琼脂克隆形成能力和肝癌细胞增殖能力均受到不同程度的抑制。流式细胞结果表明,干扰HOXA2基因表达能够减缓细胞周期进展,使肝癌细胞阻滞于G1期,并且S期减少。Annexin V和PI双染色细胞凋亡实验表明,干扰HOXA2基因能促进肝癌细胞凋亡。结论HOXA2基因可以较显著影响肝癌细胞增殖和凋亡。Objective To study the role of homeobox A2（HOXA2） gene in cell growth,cellcycle and apoptosis of hepatoma cells, and discuss the feasibility that HOXA2 gene would be a potential therapy target of hepatoma. Methods Real-time quantitative polymerase chain reaction and revers transcriptional polymerase chain reaction were performed to examine HOXA2 expression in tumorous and non-tumorous tissue of patients was matched with liver cancer. siRNAs were chemically synthesized to interference HOXA2 in PLC/PRF/5 and MHCC-97L cells. Growth curve and soft-agar colony formation assay were performed to evaluate cell growth, and cell cycle and apop-tosis were analyzed by flow cytometry. Results HOXA2 was upregulated in HCC samples compared with matched non-tumor tissues（P＆lt;0.05）. Two siRNAs against HOXA2 gene were designed and synthesized to specially knock-down HOXA2 in PLC/PRF/5 and MHCC-97L cells. HOXA2 knockdown inhibited cellular growth and soft-agar colony formation in PLC/PRF/5 and MHCC-97L cells. And Fowcytometry analysis revealed that HOXA2 knock-down by RNA interference could result in G1 arrest and S decrease and promoted cellular apoptosis. ConclusionHOXA2 gene has an important role in cell growth of hepatoma cells.

关 键 词：RNA干扰 同源异型盒基因 肝癌 细胞周期 细胞凋亡 

分 类 号：Q291[生物学—细胞生物学]