Evaluation of the chemosensitivity of head and neck cancer cells based on the diverse function of mutated-p53.
Evaluation of the chemosensitivity of head and neck cancer cells based on the diverse function of mutated-p53.
复制标题
基于突变p53的多样化功能评估头颈癌细胞的化疗敏感性。
DOI：
10.3892/ijo.22.2.383
复制
发表时间：
2003
影响因子：
5.2
通讯作者：
T. Fujibayashi
中科院分区：
文献类型：
--
作者：
Y. Shinagawa;H. Kawamata;F. Omotehara;K. Nakashiro;M. Hoque;T. Furihata;H. Horiuchi;Yataka Imai;T. Fujimori;T. Fujibayashi
p53基因的治疗前评估对头颈部恶性肿瘤的治疗具有重要意义。而对p53基因的分析一般采用免疫组化、聚合酶链反应-单链构象多态性（PCR-SSCP）和直接测序等方法。现在需要哺乳动物细胞中p53转录活性的功能分析系统。我们开发了一个癌细胞中p53转录活性的功能分析系统。我们使用了两个人头颈癌细胞系，HSG（Asn 30 Ser）和TYS（Asp 281 His），并携带突变的p53基因，和人骨肉瘤细胞系，Saos-2作为对照。我们用含有p53靶基因（p21 waf 1、BAX、MDM 2、p53 AIP 1或p53 AIP 1A）启动子序列的荧光素酶报告质粒转染这些细胞。用化疗药物处理细胞后，测量荧光素酶活性的变化。在HSG细胞中，没有一个靶基因启动子被化疗药物激活。在TYS细胞中，p21 waf 1启动子被化疗药物显著激活，而Bax和p53 AIP 1启动子未被激活。TYS细胞中的这种类型的突变p53防止DNA损伤导致的细胞死亡，并且可能通过DNA损伤治疗积累遗传改变并加速细胞的恶性进展。因此，分析突变型p53的不同功能可能有助于确定治疗策略，特别是头颈部癌患者的化疗和放疗策略。
Pre-therapeutic evaluation of p53 gene is very important for treating patients with head and neck cancer. However, the analysis for p53 gene has generally been done by immunohistochemistry, polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) and direct sequencing. Functional analysis system for p53 transcriptional activity in mammalian cells is now required. We developed a functional analysis system for p53 transcriptional activity in cancer cells. We used two human head and neck cancer cell lines harboring mutated p53 gene, HSG (Asn30Ser) and TYS (Asp281His), and a human osteosarcoma cell line, Saos-2 as a control. We transfected these cells with luciferase reporter plasmids containing promoter sequence of p53 target genes (p21waf1, BAX, MDM2, p53AIP1 or PUMA). After treating the cells with chemotherapeutic drugs, alteration of the luciferase activity was measured. In HSG cells, none of the target gene promoters was activated by treatment with chemotherapeutic drugs. In TYS cells, p21waf1 promoter was markedly activated by treatment with chemotherapeutic drugs, but Bax and p53AIP1 promoter was not activated. This type of mutated-p53 in TYS cells prevents cell death from DNA damage, and probably accumulates genetic alterations and accelerates the malignant progression of the cells by DNA damaging therapy. Thus, analysis for the diverse function of mutated-p53 may help to determine the therapeutic strategy, especially for chemotherapy and radiation in the individual patients with head and neck cancer.