CT120反义寡核苷酸抑制肺腺癌细胞A549的生长

Antisense oligonecleotides of CT120 inhibit the growth of lung adenocarcinoma cell line A549

目的探讨CT120基因与肺腺癌细胞A549生长、分化的关系。方法构建含有CT120基因的反义表达载体(命名为pcDNA3.1-CT120),并将该载体转染肺腺癌细胞A549,应用G418筛选,获得了稳定表达CT120反义基因的克隆(命名为pcDNA3.1-CT120-A549)。RT-PCR和Western blot检测CT120表达。流式细胞仪和软琼脂集落形成实验分析细胞生长,同时用RT-PCR检测P53、CyclinD1和CDK4的表达。结果成功构建了含有CT120基因的反义表达载体,而且该载体能够有效抑制CT120在肺腺癌细胞A549中的表达。抑制CT120基因的表达能够抑制细胞的生长,促进细胞凋亡。在稳定表达CT120反义基因的克隆中P53表达明显上调,CyclinD1和CDK4表达明显下调。结论反义寡核苷酸抑制CT120的表达有可能是肺癌基因治疗的一个新的靶点。

Objective To clarify the relationship between CT120, a novel human plasma membrane-associated gene, and the proliferation of lung adenocarcinoma cell line A549. Methods Expression vector（ pcDNA3. 1 ） containing antisense oligonucleotides of CT120 was constructed and transfected into the adenocarcinoma cell line A549. RT-PCR and Western blot detected the expression of CT120. Meantime, flow cytometry and soft agarose colony formation were applied for cell proliferation, and P53, CyclinD1 and CDK4 were detected by RT-PCR. Results pcDNA3. 1 containing antisense oligonucleotides of CT120 was successfully constructed and inhibited the expression of CT120 effectively by RT-PCR and Western blot. The expression of P53 was up-regulated and the expression of CyclinD1 and CDK4 were down-regulated. Conclusion The down-regulatio gonucleotides technique may be a potential drug target for treatment of lung n of CT120 expression by antisense olicancer.