敲低蛋白磷酸酶2A的癌性抑制因子表达对下咽癌细胞增殖、迁移的影响

Effect of CIP2A knockdown on proliferation and migration of hypopharyngeal cancer cells

目的通过shRNA敲低下咽癌FaDu细胞(简称FaDu细胞)蛋白磷酸酶2A(protein phosphatase 2A,PP2A)的癌性抑制因子(cancerous inhibitor of protein phosphatase 2A,CIP2A)的表达,了解其在下咽癌发生发展中的作用。方法设计特异性shRNA序列,包装慢病毒并转染FaDu细胞,特异性敲低CIP2A的表达,分别通过RT-PCR和Western blot检测CIP2A mRNA和CIP2A蛋白的表达情况。结果(1)shRNA敲低FaDu细胞中CIP2A后,实验组CIP2AmRNA和CIP2A蛋白的表达量均较空白组显著降低。(2)细胞克隆和CCK8实验结果显示,实验组细胞较空白组细胞增殖能力明显下降(t=50.86,P<0.01;t=12.406,P<0.001)。细胞划痕实验结果显示,实验组细胞横向迁移能力较空白组细胞明显下降。Transwell实验结果显示,实验组细胞纵向迁移能力较空白组细胞明显下降(t=40.038,P<0.01;t=12.247,P<0.001)。结论敲低FaDu细胞CIP2A后细胞增殖和迁移能力均下降,表明CIP2A参与调控下咽癌细胞的生物学行为,可作为潜在的抗癌靶点。

OBJECTIVE The expression of cancerous inhibitor of protein phosphatase 2A(CIP2A)in hypopharyngeal carcinoma FaDu cells(FaDu cells)was reduced by shRNA to understand its role in the occurrence and development of hypopharyngeal carcinoma.METHODS Specific shRNA sequence was designed,lentivirus was packaged and transfected into hypopharyngeal carcinoma FaDu cells,and CIP2A expression was specifically knocked down.The expression of CIP2A was detected by RT-PCR and Western blot.RESULTS 1.After shRNA knocked down CIP2A in FaDu cells,the CIP2A mRNA expression in the experimental group(CIP2A knocked down group)was significantly lower than that in the blank group,and the CIP2A protein expression in the experimental group was also significantly lower than that in the blank group.2.Cell cloning and CCK8 experiments showed that the cell proliferation ability of the experimental group was significantly decreased compared with that of the blank group(t=50.86,P<0.01;t=12.406,P<0.001);The results of cell scratch test showed that the transverse migration ability of the experimental group was significantly decreased compared with the blank group,and the longitudinal migration ability of the experimental group was significantly decreased compared with the blank group by Transwell test(t=40.038,P<0.01;t=12.247,P<0.001).CONCLUSION After knockdown CIP2A expression in hypopharyngeal carcinoma FaDu cells,the proliferation and migration ability of hypopharyngeal cancer cells decreased,suggesting that CIP2A is involved in regulating the biological behavior of hypopharyngeal cancer cells and can be used as a potential anticancer target.