抑制Plk1表达对食管癌细胞化疗敏感性的影响

The Effect of Sirna-mediated Plk1 Depletion on Chemo-sensitivity in Esophageal Cancer Cells

研究食管癌细胞中Plk1表达被抑制后对化疗药物敏感性的影响,为进一步研究开发基于Plk1食管癌分子靶向治疗奠定基础。化学合成法合成特异性的短链Plk1siRNA,脂质体转染法将短链siRNA分别导入3代表性的食管癌细胞系;采用RT-PCR和蛋白质印迹法确认siRNA对Plk1表达的抑制效果;MTT分析观察抑制Plk1表达对食管癌细胞化疗药物敏感性的影响。半定量RT-PCR和蛋白质印迹分析结果表明,合成的特异性短链Plk1siRNA在mRNA和蛋白质水平上均能高效、特异性地抑制所使用的3代表性食管癌细胞系中Plk1的表达,抑制程度约90%。MTT分析结果表明,siRNA抑制Plk1表达后,能显著提高3食管癌细胞系对化疗药物阿霉素和顺铂的敏感性。由此证实,siRNA介导的Plk1表达抑制能显著提高食管癌细胞对化疗药物阿霉素和顺铂的敏感性。

It aimed at investigate the effect of siRNA-mediated Plk1 depletion on chemo-sensitivity in esophageal cancer cells. Small interfering RNA（siRNA）was used to specifically deplete Plk1 in esophageal cancer cells. RT-PCR and immunoblotting analysis were used to verify the inhibitory effect of siRNA against Plk1. MTT assay was conducted to determine the effects of Plk1 depletion on the chemo-sensitivity of esophageal cancer cells. Synthetic siRNA duplexes against Plk1 were introduced into three esophageal cancer cell lines,which subsequently resulted in a significant inhibition in Plk1 expression in the cells. MTT assay revealed that the targeted depletion of Plk1 significantly enhanced the chemo-sensitivity to chemotherapeutic agents including adriamycin and eisplatin in all three esophageal cancer cell lines studied. It proved that siRNA-mediated Plk1 depletion can significantly enhanced the ehemo-scnsitivity of esophageal cancer cells to chemotherapeutic agents such as adriamycin and eisplatin.