Inhibition of cyclin D1 expression by cyclin D1 shRNAs in human oral squamous cell carcinoma cells is associated with increased cisplatin chemosensitivity
Inhibition of cyclin D1 expression by cyclin D1 shRNAs in human oral squamous cell carcinoma cells is associated with increased cisplatin chemosensitivity
复制标题
人口腔鳞状细胞癌细胞中细胞周期蛋白 D1 shRNA 抑制细胞周期蛋白 D1 表达与顺铂化疗敏感性增加相关
DOI：
10.1002/ijc.23964
复制
发表时间：
2009-01-15
影响因子：
6.4
通讯作者：
Zhang, Ping
中科院分区：
文献类型：
--
作者：
Zhou, Xiaojian;Zhang, Zhiyuan;Zhang, Ping
细胞周期蛋白D1是一种众所周知的细胞周期调节因子。最近，它的促生存功能已在几种肿瘤中被发现。由于细胞周期蛋白D1的表达增加是口腔鳞状细胞癌（OSCC）的常见事件，并已与顺铂耐药，我们调查，如果细胞周期蛋白D1抑制可以增加顺铂化疗敏感性的OSCC。制备5个细胞周期蛋白D1的shRNA，并选择3个用于后续实验。通过MTT测定法测定顺铂的IC 50值。观察顺铂诱导的细胞凋亡和细胞周期阻滞。建立肿瘤移植模型，检测体内细胞周期蛋白D1沉默后Tca/顺铂对顺铂的敏感性。核因子-κ B（NF-κ B）和细胞周期蛋白依赖性激酶4（CDK 4）在细胞周期蛋白D1介导的顺铂耐药中的作用也进行了研究。最有效的shRNA导致84.51%的cyclin D1蛋白水平的敲低。在用两种最有效的shRNAs转染后，顺铂的IC 50从5.88 μ g/ml降低到1.36 μ g/ml和24.47 μ g/ml，尽管细胞周期蛋白D1的过表达使OSCC细胞对顺铂治疗更具抗性（IC 50从6.43 μ g/ml增加到12.24 μ g/ml）。这种降低的IC 50与在细胞周期蛋白D1敲低细胞中顺铂诱导的NF-κ B活性下调相关，并且与CDK 4功能无关。体内肿瘤移植模型也证实了cyclin D1 shRNA在OSCC中的顺铂增敏作用。TUNEL检测证实了顺铂在细胞周期蛋白D1敲低细胞中诱导的凋亡增加。细胞周期蛋白D1可能是口腔鳞癌患者未来治疗的一个重要靶点。(c)2008威利利斯公司
Cyclin D1 is a well-known cell cycle regulator. Recently, its pro-survival function has been revealed in several tumors. Because increasing expression of cyclin D1 is a common event in oral squamous cell carcinoma (OSCC) and has been correlated with cisplatin resistance, we investigated if cyclin D1 inhibition could increase cisplatin chemosensitivity of OSCC. Five cyclin D1 shRNAs were prepared and 3 were selected for subsequent experiments. IC50 values for cisplatin were determined by an MTT assay. Cisplatin-induced apoptosis and cell cycle block were investigated. A tumor transplantation model was generated to examine the cisplatin sensitivity of Tca/cisplatin after in vivo cyclin D1 silencing. The role of nuclear factor-kappa B (NF-kappa B) and cyclin-dependent kinase 4 (CDK4) in cyclin D1-mediated cisplatin resistance was also examined. The most effective shRNA resulted in 84.51% knockdown of cyclin D1 protein level. After the transfection with the 2 most effective shRNAs, the cisplatin IC50 decreased from 5.88 mu g/ml to 1.36 mu g/ml and 24.47 mu g/ml, although overexpression of cyclin D1 rendered OSCC cells more resistant to cisplatin treatment (IC50 increased from 6.43 mu g/ml to 12.24 mu g/ml). This decreasing IC50 was correlated with the down-regulation of cisplatin-induced NF-kappa B activity in cyclin D1 knockdown cells, and was independent of CDK4 function. In vivo tumor transplantation models also confirmed a cisplatin-sensitizing effect of cyclin D1 shRNA in OSCC. A TUNEL assay validated the increase in apoptosis as induced by cisplatin in cyclin D1 knockdown cells. Cyclin D1 may be an important target for future therapy in patients with OSCC. (c) 2008 Wiley-Liss, Inc.