Estrogen regulation in endometrial carcinoma Ishikawa cells LRP16 gene expression and its significance.docVIP

 PAGE \* MERGEFORMAT 10 Estrogen regulation in endometrial carcinoma Ishikawa cells LRP16 gene expression and its significance Author: Meng Yuan-guang, South Korea to the East, Huang Ke, Wu Zhiqiang, Zhao Yali, Mu Yi-ming, SONG Lei [Keywords:] LRP16 gene; estrogen; Ishikawa cells; E Expression regulation of LRP16 gene by 17β estradiol and its significance in human endometrial cancer Ishikawa cells [Abstract] AIM: To explore the regulation of LRP16 gene expression by 17β estradiol (E2) in ERα positive human endometrial cancer Ishikawa cell line, and to investigate the effect of LRP16 overexpression on the proliferative potential and invasive growth of Ishikawa and the possible molecular mechanism. METHODS: The LRP16 mRNA level in Ishiwaka cells was determined by Northern blot analysis. The relative luciferase activity was measured using Dualluciferase reporter assay system. The effect of LRP16 overexpression on Ishiwaka proliferation was examined by the Trypan Blue exclusion method. The invasiveness of Ishikawa was evaluated by using the Matrigelcoated Transwell assay. The protein levels in Ishikawa were determined by Western blot analysis. In addition, the Ecadherin mRNA level was also examined by Nothern blot. RESULTS: 17β E2 induced an increase in LRP16 mRNA levels in Ishikawa cells, whereas, its pure antagonist, ICI 182 780 reduced the LRP16 mRNA level. Ectopic ERα transfection in Ishikawa increased the expression of LRP16 gene. The significant increase of the relative luciferase activity in Ishikawa cells cotransfected by pGL3S5 and ERα plasmids was observed compared with that in the control cells transfected by pGL3S5 alone. Stimulative effect of LRP16 overexpression on the proliferation of Ishikawa cells was not observed in this study. Thirtypersent increase of the invasive capacity was observed in Ishikawa cells with LRP16 overexpression. The mRNA and protein levels of Ecadherin gene was nearly 3fold decreased in Ishikawa cells with the overexpre