Sato M, Kawamata H, Harada K, Nakashiro K, Ikeda Y, Gohda H, Yoshida H, Nishida T, Ono K, Kinoshita M, Adachi M
Second Department of Oral and Maxillofacial Surgery, Tokushima University School of Dentistry, Japan.
Cancer Lett. 1997 Jan 30;112(2):181-9. doi: 10.1016/s0304-3835(96)04581-8.
It has been found by PCR-SSCP analysis and direct DNA sequencing that a human salivary adenosquamous carcinoma-forming cell line, TYS, has a mutant p53 gene at codon 281Asp-->His. When TYS cells were treated with a differentiation-inducing agent, vesnarinone, cellular proliferation was significantly inhibited on the basis of MTT assay. In addition, it has been found by Northern blotting and/or immunoblotting that expression of p21WAF1 and transforming growth factor-beta (TGF-beta) is up-regulated by treating TYS cells with vesnarinone. TGF-beta 1 alone also induced p21WAF1 expression in TYS cells. Moreover, it has been shown by ELISA that the treatment of TYS cells with vesnarinone results in the enhanced generation of latent TGF-beta 1. The expression of TGF-beta receptor (T beta R), including T beta R-I, T beta R-II and T beta R-III, on TYS cells was detected by affinity cross-linking using 125I-TGF-beta 1 and addition of active TGF-beta 1 into serum-free culture medium inhibited the growth of TYS cells in a concentration-dependent manner. These findings suggest that vesnarinone might directly induce expression of p21WAF1 gene in TYS cells, the product of which may be associated with the inhibition of cell growth and induce differentiation.
通过聚合酶链反应-单链构象多态性分析（PCR-SSCP）和直接DNA测序发现，人涎腺腺鳞癌形成细胞系TYS在密码子281处存在p53基因突变（天冬氨酸Asp→组氨酸His）。当用分化诱导剂维斯纳林酮处理TYS细胞时，基于MTT法检测，细胞增殖受到显著抑制。此外，通过Northern印迹法和/或免疫印迹法发现，用维斯纳林酮处理TYS细胞可上调p21WAF1和转化生长因子-β（TGF-β）的表达。单独的TGF-β1也可诱导TYS细胞中p21WAF1的表达。此外，通过酶联免疫吸附测定（ELISA）表明，用维斯纳林酮处理TYS细胞可导致潜伏性TGF-β1的生成增加。使用125I-TGF-β1通过亲和交联检测TYS细胞上TGF-β受体（TβR）的表达，包括TβR-I、TβR-II和TβR-III，并且向无血清培养基中添加活性TGF-β1可浓度依赖性地抑制TYS细胞的生长。这些发现表明，维斯纳林酮可能直接诱导TYS细胞中p21WAF1基因的表达，其产物可能与细胞生长抑制和诱导分化有关。
