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背景与目的:30%NSCLC存在HER2/neu过表达,与肿瘤发生、转移、肿瘤血管形成、抗凋亡及化疗耐药等有关。本文通过RNA干涉抑制肺腺癌细胞SPC-A-1 HER2/neu过表达,观察肿瘤细胞周期、增殖及集落形成能力的变化。方法:构建针对HER2/neu的siRNA重组质粒,稳定转染SPC-A-1,通过RT-PCR和W esternB lot检测HER2/neu表达;FCM分析细胞周期;MTT法观察细胞增殖,绘制细胞生长曲线;平板集落形成实验检测肿瘤细胞的集落形成能力。结果:成功构建了HER2/neu的siRNA重组质粒,稳定转染靶细胞后可显著降低HER2/neu表达;与亲代细胞相比,G0/G1期细胞增加17.1%,S期细胞减少12.8%;细胞生长速度减慢,集落S1抑制率达到49.0%。结论:针对HER2/neu的siRNA可以显著降低肺腺癌细胞过表达HER2/neu,肿瘤细胞阻滞于G0/G1期,造成细胞增殖减慢,集落形成能力明显下降。因此,siRNA对过表达HER2/neu肺癌的治疗具有潜在应用价值。
BACKGROUND & AIM: HER2 / neu overexpression is present in 30% of NSCLC and is associated with tumorigenesis, metastasis, tumor angiogenesis, anti-apoptosis and chemoresistance. In this paper, RNA interference was used to inhibit the overexpression of SPC-A-1 HER2 / neu in lung adenocarcinoma cells. The cell cycle, proliferation and colony formation ability were observed. Methods: The recombinant plasmid targeting HER2 / neu was constructed and stably transfected into SPC-A-1. The expression of HER2 / neu was detected by RT-PCR and Western blot. The cell cycle was analyzed by FCM. The cell proliferation was observed by MTT assay. ; Colony formation assay test colony formation ability of tumor cells. Results: The siRNA recombinant plasmid of HER2 / neu was successfully constructed, and HER2 / neu expression was significantly reduced after stable transfection of target cells. Compared with parental cells, cells in G0 / G1 phase increased by 17.1% and cells in S phase decreased by 12.8% Growth slowed down, colonies S1 inhibition rate reached 49.0%. Conclusion: siRNA targeting HER2 / neu can significantly reduce the overexpression of HER2 / neu in lung adenocarcinoma cells. The arrest of tumor cells in G0 / G1 phase resulted in slowed cell proliferation and decreased colony-forming ability. Therefore, siRNA has potential value in the treatment of overexpressing HER2 / neu lung cancer.