Hastings Randolph H, Montgrain Philippe R, Quintana Rick, Rascon Yvette, Deftos Leonard J, Healy Erin
Department of Anesthesiology, Veterans Affairs San Diego Healthcare System, San Diego, CA 92161, USA.
Am J Physiol Lung Cell Mol Physiol. 2009 Oct;297(4):L578-85. doi: 10.1152/ajplung.90560.2008. Epub 2009 Jul 24.
Parathyroid hormone-related protein (PTHrP), a paraneoplastic protein expressed by two-thirds of human non-small cell lung cancers, has been reported to slow progression of lung carcinomas in mouse models and to lengthen survival of patients with lung cancer. This study investigated the effects of ectopic expression of PTHrP on proliferation and cell cycle progression of two human lung adenocarcinoma cell lines that are normally PTHrP negative. Stable transfection with PTHrP decreased H1944 cell DNA synthesis, measured by thymidine incorporation, bromodeoxyuridine uptake, and MTT proliferation assay. A substantial fraction of PTHrP-positive cells was arrested in or slowly progressing through G1. Cyclin D2 and cyclin A2 protein levels were 60-70% lower in PTHrP-expressing cells compared with control cells (P < 0.05, N = 3 independent clones per group), while expression of p27(Kip1), a cyclin-dependent kinase inhibitor, was increased by 35 +/- 9% (mean +/- SE, P < 0.05) in the presence of PTHrP. Expression of other cyclins, including cyclins D1 and D3, and cyclin-dependent kinases was unaffected by PTHrP. PTHrP did not alter the phosphorylation state of Rb, but decreased cyclin-dependent kinase (CDK) 2-cyclin A2 complex formation. Ectopic expression of PTHrP stimulated ERK phosphorylation. In MV522 cells, PTHrP had similar effects on DNA synthesis, cyclin A2 expression, pRb levels, CDK2-cyclin A2 association, and ERK activation. In summary, PTHrP appears to slow progression of lung cancer cells into S phase, possibly by decreasing activation of CDK2. Slower cancer cell proliferation could contribute to slower tumor progression and increased survival of patients with PTHrP-positive lung cancer.
甲状旁腺激素相关蛋白(PTHrP)是一种由三分之二的人类非小细胞肺癌表达的副肿瘤蛋白,据报道,在小鼠模型中它可减缓肺癌的进展,并延长肺癌患者的生存期。本研究调查了PTHrP异位表达对两种正常情况下PTHrP阴性的人肺腺癌细胞系增殖和细胞周期进程的影响。通过胸苷掺入、溴脱氧尿苷摄取和MTT增殖试验测量,用PTHrP进行稳定转染可降低H1944细胞的DNA合成。相当一部分PTHrP阳性细胞停滞在G1期或在G1期缓慢进展。与对照细胞相比,表达PTHrP的细胞中细胞周期蛋白D2和细胞周期蛋白A2的蛋白水平降低了60 - 70%(P < 0.05,每组N = 3个独立克隆),而细胞周期蛋白依赖性激酶抑制剂p27(Kip1)的表达在有PTHrP存在时增加了35 +/- 9%(平均值 +/- 标准误,P < 0.05)。包括细胞周期蛋白D1和D3以及细胞周期蛋白依赖性激酶在内的其他细胞周期蛋白的表达不受PTHrP影响。PTHrP没有改变Rb的磷酸化状态,但减少了细胞周期蛋白依赖性激酶(CDK)2 - 细胞周期蛋白A2复合物的形成。PTHrP的异位表达刺激了ERK磷酸化。在MV522细胞中,PTHrP对DNA合成、细胞周期蛋白A2表达、pRb水平、CDK2 - 细胞周期蛋白A2结合以及ERK激活具有相似的作用。总之,PTHrP似乎通过降低CDK2的激活来减缓肺癌细胞进入S期的进程。癌细胞增殖减慢可能有助于减缓肿瘤进展,并提高PTHrP阳性肺癌患者的生存率。