Sweeney Christopher, Li Lang, Shanmugam Rajasubramaniam, Bhat-Nakshatri Poornima, Jayaprakasan Vetrichelvan, Baldridge Lee Ann, Gardner Thomas, Smith Martin, Nakshatri Harikrishna, Cheng Liang
Department of Medicine, Indiana University, Indianapolis, Indiana 46202, USA.
Clin Cancer Res. 2004 Aug 15;10(16):5501-7. doi: 10.1158/1078-0432.CCR-0571-03.
The transcription factor nuclear factor-kappaB (NF-kappaB) promotes the production of angiogenic, antiapoptotic, and prometastatic factors that are involved in carcinogenesis.
Electromobility gel shift assays were used to evaluate NF-kappaB DNA binding in vitro. The functional relevance of NF-kappaB DNA binding was assessed by both cDNA array analyses and proliferation assays of prostate cancer cells with and without exposure to an NF-kappaB inhibitor, parthenolide. Immunohistochemistry staining for the p65 NF-kappaB subunit was used to determine the frequency and location of NF-kappaB in 97 prostatectomy specimens. The amount of staining was quantified on a 0-3+ scale.
An electromobility gel shift assay confirmed the presence of NFkappaB DNA binding in all four prostate cancer cell lines tested. The binding was inhibited by parthenolide, and this agent also decreased multiple gene transcripts under the control of NF-kappaB and inhibited proliferation of prostate cancer cells. The staining results revealed overexpression of p65 in the prostatic intraepithelial neoplasia and cancer compared with the benign epithelium. Specifically, there was a predominance of 1+ and 2+ with no 3+ staining in benign epithelium, whereas there was only 2+ and 3+ staining (30 and 70%, respectively) in the cancerous areas. These differences were statistically different. There was no correlation with tumor grade or stage.
NF-kappaB is constitutively activated in prostate cancer and functionally relevant in vitro. Immunohistochemistry of human prostatectomy specimens demonstrated overexpression of the active subunit of NF-kappaB, p65, and that this occurs at an early stage in the genesis of prostate cancer. This work supports the rationale for targeting NF-kappaB for the prevention and/or treatment of prostate cancer.
转录因子核因子-κB（NF-κB）促进血管生成、抗凋亡和促转移因子的产生，这些因子参与致癌过程。
采用电泳迁移率凝胶移位分析（EMSA）在体外评估NF-κB与DNA的结合。通过cDNA阵列分析以及对暴露和未暴露于NF-κB抑制剂小白菊内酯的前列腺癌细胞进行增殖分析，评估NF-κB与DNA结合的功能相关性。使用p65 NF-κB亚基的免疫组织化学染色来确定97例前列腺切除标本中NF-κB的频率和位置。染色量按0-3+级进行量化。
电泳迁移率凝胶移位分析证实，在所测试的所有四种前列腺癌细胞系中均存在NF-κB与DNA的结合。小白菊内酯可抑制这种结合，并且该药物还可降低NF-κB控制下的多种基因转录本，并抑制前列腺癌细胞的增殖。染色结果显示，与良性上皮相比，前列腺上皮内瘤变和癌组织中p65表达上调。具体而言，良性上皮中主要为1+和2+染色，无3+染色，而癌组织中仅2+和3+染色（分别为30%和70%）。这些差异具有统计学意义。与肿瘤分级或分期无关。
NF-κB在前列腺癌中持续激活且在体外具有功能相关性。人前列腺切除标本的免疫组织化学显示，NF-κB的活性亚基p65表达上调，且这发生在前列腺癌发生的早期阶段。这项工作支持了将NF-κB作为预防和/或治疗前列腺癌靶点的理论依据。
