RNA干扰抑制CD59对非小细胞肺癌GLC-P细胞株增殖的影响

Effect of RNA interference of CD59 gene on proliferation of non-small cell lung cancer cell line GLC-P in vitro

目的运用RNAi技术抑制CD59基因的表达,观察其对非小细胞肺癌细胞株GLC-P增值、凋亡的影响。方法合成可抑制CD59基因的片段,并运用RNA干扰技术,构建重组质粒,通过脂质体转染法将重组质粒转染至非小细胞肺癌细胞株GLC-P,并筛选出稳定表达细胞后,采用PCR、MTT、ELISA法检测该稳定表达细胞株的增值、凋亡所受到的影响。结果干扰后的GLC-P细胞株中CD59 m RNA表达量较未干扰组显著降低(P<0.05),同时MTT、ELISA实验检测显示肺癌GLC-P细胞增殖能力降低,可诱导肺癌细胞的凋亡。结论肺癌患者肿瘤细胞中存在CD59高表达,抑制CD59基因表达的同时可抑制GLC-P细胞的增殖能力并诱导细胞凋亡,为非小细胞肺癌的靶向治疗提供了新靶点、新思路。

Objective To investigate the effect of CD59 gene inhibition mediated by RNA interference on the proliferation and apoptosis of non-small cell lung cancer（NSCLC） GLC-P cells in vitro. Methods Recombinant plasmids for RNA interference of CD59 gene were constructed and transfected into GLC- P cells via lipofectamine 2000. The stably transfected cells were examined with real- time RT- PCR, MTT assay and enzyme- linked immunosorbent assay to investigate the changes in cell proliferation and apoptosis. Results Compared with the control cells, the cells transfected with CD59- si RNA showed significantly decreased expression levels of CD59 m RNA（P〈0.05） and significantly inhibited cell proliferation. Conclusions CD59 gene is highly expressed in NSCLC and RNA interference-mediated CD59 silencing can strongly inhibit the proliferation and induce apoptosis in GLC-P cells, which shed light on a potentially new target for targeted gene therapy of NSCLC.