Sauter E R, Herlyn M, Liu S C, Litwin S, Ridge J A
Department of Surgery, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Clin Cancer Res. 2000 Feb;6(2):654-60.
Local recurrence of squamous cell cancer (SCC) causes high morbidity and is often readily accessible, making such patients potential candidates for gene therapy. Cyclin D1 (CD1), critical in the G1-S transition in the cell cycle, is amplified in 20-50% and overexpressed in up to 80% of head and neck SCC. Our earlier studies indicated that CD1 expression increased with progression from low grade to high grade dysplasia, and that treatment of established tumors with antisense cyclin D1 (AS-cyclin D1) led to tumor regression during a one week evaluation period. We hypothesized that: 1) CD1 expression increases with disease progression to advanced SCC, and 2) AS-cyclin D1 therapy would lead to prolonged tumor regression in a xenograft model of human SCC. CD1 expression, evaluated by immunostain in 30 stage III/IV head and neck SCC, increased in the basal layer from normal-dysplasia (P = 0.06) and from dysplasia-carcinoma (P = 0.004). In the germinative layer CD1 expression increased from dysplasia-carcinoma (P = 0.002) but not from normal-dysplasia. Western blotting of eight SCC and two transformed keratinocyte cell lines demonstrated CD1 overexpression in 8/10 (80%) lines. An 11th cell line (A431) had previously been shown to overexpress cyclin D1. 8/9 (89%) cell lines overexpressing CD1 formed tumors in immunodeficient mice, whereas 0/2 cell lines without CD1 overexpression formed a tumor. Three established SCCs, one fast growing, one with moderate growth rate (with CD1 overexpression) and one slow growing (without increased CD1), shrank significantly for 2-4 weeks after AS-cyclin D1 treatment, while tumors transduced with control vector grew. Cyclin D1 expression increases in frequency with disease progression, and antisense cyclin D1 was effective in a xenograft model of human cancer, independent of tumor growth rate.
鳞状细胞癌（SCC）的局部复发会导致高发病率，且通常易于治疗，这使得此类患者成为基因治疗的潜在候选对象。细胞周期蛋白D1（CD1）在细胞周期的G1-S转换中起关键作用，在20%-50%的头颈部SCC中发生扩增，在高达80%的头颈部SCC中过表达。我们早期的研究表明，CD1表达随着从低级别发育异常到高级别发育异常的进展而增加，并且用反义细胞周期蛋白D1（AS-细胞周期蛋白D1）治疗已形成的肿瘤在一周的评估期内导致肿瘤消退。我们假设：1）CD1表达随着疾病进展至晚期SCC而增加；2）AS-细胞周期蛋白D1治疗将导致人SCC异种移植模型中的肿瘤长期消退。通过免疫染色评估30例III/IV期头颈部SCC中的CD1表达，其在基底细胞层中从正常-发育异常（P = 0.06）以及从发育异常-癌（P = 0.004）增加。在生发层中，CD1表达从发育异常-癌增加（P = 0.002），但从正常-发育异常未增加。对8种SCC和2种转化的角质形成细胞系进行蛋白质免疫印迹分析显示，8/10（80%）的细胞系中CD1过表达。第11种细胞系（A431）先前已被证明过表达细胞周期蛋白D1。8/9（89%）过表达CD1的细胞系在免疫缺陷小鼠中形成肿瘤，而2种未过表达CD1的细胞系中0/2形成肿瘤。3个已形成的SCC，1个生长迅速，1个生长速度中等（有CD1过表达），1个生长缓慢（无CD1增加），在AS-细胞周期蛋白D1治疗后2-4周显著缩小，而用对照载体转导的肿瘤生长。细胞周期蛋白D1表达频率随疾病进展增加，并且反义细胞周期蛋白D1在人癌异种移植模型中有效，与肿瘤生长速度无关。
