Inhibitory effects on tumor invasion and metastasis induced by transfection of anti-sense E1AF,an ets-oncogene family transcription factor.

E1AF是一个新发现的人类Ets家族转录因子。我们已经报道，E1AF可以上调基质金属蛋白酶(MMPs)基因的转录，并赋予人癌细胞侵袭性表型。HSC3是一种口腔鳞状细胞癌来源的细胞系，其E1AF和基质金属蛋白酶-1、-9基因的高表达与口腔鳞癌的侵袭潜能有关。我们构建了E1AF反义表达载体，用该载体转染HSC3细胞，获得了表达E1AF反义RNA的HSC3AS细胞，HSC3AS的m、-3和-9呈下降趋势。此外，HSC3AS在体外三维RAFT培养和体内移植到裸鼠体内的侵袭能力较低。这些结果表明，反义E1AF通过下调基质金属蛋白酶基因来抑制肿瘤的侵袭。因此，E1AF被认为与癌细胞的恶性潜能高度相关，然而，在体内…中E1AF的表达与癌细胞恶性程度的关系却知之甚少更多的肿瘤。采用逆转录聚合酶链式反应、Southern杂交和原位杂交技术对27例口腔鳞状细胞癌标本进行检测，并与临床病理参数进行比较。27例患者中15例出现E1AF。17例侵袭性鳞癌S中有13例表达E1AFmRAN，而未表达E1AFmRAN的鳞癌S多呈膨胀性生长。在有淋巴结转移的病例中，E1AF阳性口腔鳞癌的患病率增加。这些结果表明，E1AF可能通过促进肿瘤细胞的侵袭潜能而参与肿瘤的发生。Waf1/Cip1&gt是细胞周期、终末分化和凋亡的关键调控蛋白之一。其启动子被野生型P53蛋白反式激活，并且不依赖于P53。我们证明了E1AF，一个ETS相关的转录因子，通过与两个先前发现的p53反应元件附近的ETS结合位点相互作用，激活了人的p21^；Waf1/Cip1&gt；启动子。Northern印迹分析显示顺铂诱导的SiHA细胞p21^-lt；Waf1/Cip1&gt；和E1AF表达上调。瞬时表达分析表明，E1AF可在SiHA细胞中激活p21^&lt；Waf1/Cip1&gt；启动子驱动的荧光素酶报告基因。在p53缺失的Saos2骨肉瘤细胞中，p21^&lt；Waf1/Cip1&gt；启动子活性也增加，但当Ets结合位点被删除时，活性显著降低。这些结果表明，E1AF正向调控p21^&lt；Waf1/Cip1&gt；启动子转录，以响应基因毒性胁迫。较少

E1AF is a newly identified human ets-family transcription factor. We have reported that E1AF can up-requlate transcription of matrix metalloproteinase (MMP) genes and confers invasive phenotype on human cancer cells. HSC3 is an oral squamous cell carcinoma-derived cell line, and it manifests high levels of E1AF,and MMP-1 and -9 gene expression that are associated with invasive potential. We reconstructed an E1AF antisense expression vector, transfected HSC3 cells with the vector and obtained HSC3AS cells which express E1AF antisense RNA.HSC3AS showed decreasing m, -3 and -9. moreover, HSC3AS showed lower invasive potential in in vitro three-dimensional raft culture and in vivo implantation into nude mice. These results imply that transfection of antisense E1AF inhibits tumor invasion by down-regulating MMP genes.Thus, E1AF is thought to be highly correlated with malignant potentials of cancer cells, however, little is known about E1AF expression and cancer cell malignancies in in vivo … More tumors. We examined 27 oral SCC speciment using RT-PCR,Southern blot hybtidization and in situ hybridization (ISH) and compared to the clinico-pathological parameters. Among the 27 patients, E1AF was detected in 15 cases. E1AF mRAN was detected in 13 of 17 invasive SCC_s, whereas the majority of SCC_s not expressing E1AF showed an expansive growth pattern. Increased prevalence of E1AF-positive oral SCC was observed in cases with nodal metastasis. These results indicate that E1AF may be involved in cancer cell malignancies through its ability to promote invasive potentialp21^<Waf1/Cip1> is one of the key requlatory proteins in cell cycle, terminal differentiation and apoptosis. its promoter was shown to be transactivated by the wild-type p53 protein as well as in a p53-independent manner. We demonstrate that E1AF,an ets-related transcription factor, activates the human p21^<Waf1/Cip1> promoter by interacting with the ets-binding sites located close to the two previously identified p53-responsive elements. Northern blot analysis revealed that p21^<Waf1/Cip1> and E1AF were correlatively uprequlated in response to cisplatin treatment in SiHa cells. Transient expression assays demonstrated that E1AF can activate the p21^<Waf1/Cip1> promoter-driven luciferase reporter gene in SiHa cells. The p21^<Waf1/Cip1> promoter activity was also increased in p53-null Saos2 osteosarcoma cells, but was markedly reduced when the etsbinding sites were deleted. These results indicate that E1AF positively requlates transcription from the p21^<Waf1/Cip1> promoter in response to genotoxic stresses. Less