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目的观察高度癌细胞表达1(Hec1)基因对肿瘤细胞分裂周期及其肿瘤形成的影响。方法利用RT-PCR、免疫荧光和Western blot从m RNA和蛋白质表达水平分析Hec1在小鼠实体瘤细胞和正常分化细胞中表达差异;应用RNAi技术设计靶向Hec1的sh RNA序列,在体外和体内沉默S180细胞中的Hec1基因,观察体外培养S180细胞和接种S180细胞小鼠肿瘤细胞内Hec1基因表达变化。结果 RT-PCR分析表明Hec1基因的m RNA在正常组织中不表达或极少表达,而在肿瘤组织和肿瘤细胞中高表达,免疫荧光和蛋白印迹表明Hec1蛋白在体外培养的S180细胞和实体肿瘤中高度表达,在正常组织中无表达;Hec1基因sh RNA干扰后,显著抑制了S180细胞中Hec1基因的表达,sh RNA干扰对小鼠肿瘤抑制率为63%,显著抑制S180细胞的分裂增殖。结论综上所述,Hec1在肿瘤细胞的分裂与增殖中起着重要的作用。
Objective To observe the effect of highly expressed Hec1 gene on tumor cell division cycle and tumor formation. Methods The expression of Hec1 in mouse solid tumor cells and normal differentiated cells was analyzed by RT-PCR, immunofluorescence and Western blot. The sh RNA sequence targeting Hec1 was designed by RNAi technique. The expression of Hec1 in vitro and in vivo Hec1 gene was silenced in S180 cells to observe the changes of Hec1 gene expression in S180 cells and S180 cells. Results RT-PCR analysis showed that the m RNA of Hec1 gene was not expressed or rarely expressed in normal tissues, but highly expressed in tumor tissues and tumor cells. Immunofluorescence and Western blotting showed that Hec1 protein was highly expressed in S180 cells and solid tumors Highly expressed and no expression in normal tissues. After Hec1 gene sh RNA interference, the expression of Hec1 gene was significantly inhibited in S180 cells. Sh RNA interference inhibited tumor growth in mice by 63%, significantly inhibiting the proliferation and proliferation of S180 cells. Conclusion In summary, Hec1 plays an important role in tumor cell division and proliferation.