Department of Gynecology, the First Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning, PR China.
Oncol Rep. 2012 May;27(5):1461-6. doi: 10.3892/or.2012.1655. Epub 2012 Jan 26.
Adrenomedullin (AM), a potent vasodilator peptide, is present in various types of tumors. Here, we constructed short hairpin RNA (shRNA) in order to target the AM gene in vitro using RNA interference (RNAi) technology. HO8910 ovarian cancer cells were transfected, and the effects of AM on proliferation and chemosensitivity of the cells were examined. RT-PCR, real-time PCR and western blot analysis were performed to detect the AM gene and protein expression. The MTT assay was used to observe the effect of AM on proliferation and chemosensitivity of the cells. Also, the protein levels of bcl-2 and the extracellular regulated protein kinase (ERK) were evaluated by western blot analysis. We found that silencing of the AM gene inhibited the proliferation and increased the chemosensitivity of HO8910 cells, reduced the expression of AM mRNA and protein as well as downregulated bcl-2 and p-ERK expression. We, therefore, conclude that silencing of the AM gene in HO8910 ovarian cancer cells inhibited the proliferation and increased the chemosensitivity of the cells through downregulation of ERK and bcl-2 expression. Thus, anti-AM treatment together with suppression of ERK and bcl-2 expression provides a novel research approach for ovarian cancer.
肾上腺髓质素 (AM) 是一种有效的血管舒张肽,存在于多种类型的肿瘤中。在这里,我们构建了短发夹 RNA (shRNA),以便使用 RNA 干扰 (RNAi) 技术在体外靶向 AM 基因。转染 HO8910 卵巢癌细胞,并检测 AM 对细胞增殖和化疗敏感性的影响。进行 RT-PCR、实时 PCR 和 Western blot 分析以检测 AM 基因和蛋白表达。MTT 测定用于观察 AM 对细胞增殖和化疗敏感性的影响。此外,通过 Western blot 分析评估 bcl-2 和细胞外调节蛋白激酶 (ERK) 的蛋白水平。我们发现,沉默 AM 基因抑制了 HO8910 细胞的增殖并增加了细胞的化疗敏感性,降低了 AM mRNA 和蛋白的表达,并下调了 bcl-2 和 p-ERK 的表达。因此,我们得出结论,沉默 HO8910 卵巢癌细胞中的 AM 基因通过下调 ERK 和 bcl-2 的表达抑制了细胞的增殖并增加了细胞的化疗敏感性。因此,抗 AM 治疗联合抑制 ERK 和 bcl-2 的表达为卵巢癌提供了一种新的研究方法。