Study on differentiation and apoptosis-inducing therapy for head and neck cancer by vesnarinone

1.结论：1.用维司力农处理人口腔鳞癌细胞系（BHY，HN：p53野生型（exon 4 -9））和人涎腺癌细胞系（HSG，HSG-AZA 1，HSG-AZA 3：p53野生型; TYS：p53密码子281^&lt;Asp→Hls&gt;），所有细胞均出现生长抑制和G1期阻滞，其中p21^<Waf1>，p27^<Kip1>和TSC-22基因表达上调，p53基因表达下调.当维司力农口服给药于携带TYS肿瘤的裸小鼠时，肿瘤生长以及细胞分化为角质形成细胞和腺泡细胞受到显着抑制。3.维司力农对人口腔鳞状细胞癌或人涎腺腺样囊性癌有很好的治疗作用.我们从用维司力农处理的TYS细胞中分离并鉴定了人TSC-22基因。(1)TSC-22基因在人涎腺癌细胞中的下调，在体外和裸鼠中生长，引起肿瘤生长的显着加速和基因的上调抑制显着锚定非依赖性生长。(2)TSC-22蛋白在TYS细胞中的过表达导致抗癌药物（5-FU和CDDP）或辐射诱导的细胞凋亡增强。(3)TSC-22基因由3个外显子组成。转录起始位点位于TATA盒样序列下游7 bp和29 bp处。TSC-22启动子区含有转录因子如NF-kB、MyoD、AP-1、PU、C/EBP、p53、c-Myb、Sp1、NF 1或Smad的推定结合位点。转录因子Sp1和Sp3与TYS细胞中p21 Waf 1启动子中存在的维司力酮应答元件（Sp1-1和Sp1-2位点）结合. Vesarinone诱导TYS细胞中的组蛋白高乙酰化。

1. When human oral squamous cell carcinoma cell lines (BHY, HN: p53 wild type (exon4-9)) and human salivary cancer cell lines (HSG, HSG-AZA1, HSG-AZA3: p53 wild type; TYS : p53 codon 281^<Asp→Hls>) were treated with vesnarinone, inhibition of cell growth and induction of G1 arrest occurred in all of the treated cells, where up-regulation of p21^<Waf1>, p27^<Kip1> and TSC-22 genes as well as down-regulation of p53 gene were detected.2. When vesnarinone was administered per os into the nude mice bearing TYS tumors, significant inhibition of the tumor growth as well as cellular differentiation into keratinocyte and acinar cells occurred.3. Vesnarinone was very therapeutically effective for human oral squamous cell carcinoma or for human salivary adenoid cystic carcinoma.4. We isolated and characterized human TSC-22 gene from the TYS cells treated with vesnarinone.(1) Down-regulation of TSC-22 gene in human salivary cancer cells, grown in vitro and in nude mice, caused the marked acceleration of tumor growth and up-regulation of the gene inhibited significantly anchorage-independent growth.(2) Overexpression of TSC-22 protein in TYS cells resulted in the augmented induction of apotosis by anti-cancer drugs (5-FU and CDDP) or radiation.(3) TSC-22 gene was comprised of 3 exons. Transcription initiatin sites were located at 7bp and 29bp downstream from TATA box like sequence. TSC-22 promoter region contained putative binding sites for transcription factor such as NF-kB, MyoD, AP-1, PU, C/EBP, p53, c-Myb, Sp1, NF1 or Smad.5. Sp1 and Sp3 transcription factors bound to the vesnarinone-responsive element (Sp1-1 and Sp1-2 sites) present in p21Waf1 promoter in TYS cells.6. Vesnarinone induced the histone hyperacetylation in TYS cells.