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雌激素调控子宫内膜癌Ishikawa细胞中LRP16基因表达

2013-07-02 01:11 作者：孟元光，韩为东，黄柯，伍志强，赵亚力，母义明，宋磊

【关键词】 LRP16基因；雌激素；Ishikawa细胞；E
　　Expression regulation of LRP16 gene by 17βestradiol and its significance in human endometrial cancer Ishikawa cells
　　【Abstract】 AIM: To explore the regulation of LRP16 gene expression by 17βestradiol (E2) in ERαpositive 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: 17βE2 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 overexpression of LRP16, while the protein levels of MMP2, MMP9 and CD44 were not different between LRP16overexpressed Ishikawa cells and the control cells. CONCLUSION: Estrogen upregulated the LRP16 mRNA level by activation of ERα and the LRP16 expression was dependent on the estrogen in ERαpositive endometrial cancer cells. Upregulation of LRP16 did not promote the proliferation of the endometrial cancer cells, but increased its invasive potential possibly by suppressing the Ecadherin expression level.
　　【Keywords】 LRP16; estrogen; Ishikawa; Ecadherin; endometrial carcinoma
　　【摘要】 目的： 探讨子宫内膜癌Ishikawa细胞中雌激素（E2）对LRP16基因表达的调控作用，LRP16基因过表达对Ishikawa细胞增殖及侵袭生长能力的影响以及可能的分子机制. 方法： 采用Northern blot方法检测细胞中LRP16基因mRNA表达水