摘要
目的观察缺氧条件下前列腺间质细胞WPMY-1体外培养细胞膜表面生长类激素受体表达的变化。方法 WPMY-1在缺氧或有氧环境中体外培养。接种后4,8,12,24,48 h分别用RT-PCR和免疫组化法检测其细胞膜表面生长类激素受体[表皮生长因子受体(epidermal growth factor receptor,EGFR),成纤维细胞生长因子受体(fibroblast growthfactor receptor,FGFR),转化生长因子受体(transforming growth factor receptor,TGFR),胰岛素样生长因子受体(insulin-likegrowth factor receptor,IGFR),血管内皮生长因子受体(vascular endothelial growth factor receptor,VEGFR)]基因和蛋白表达。结果较有氧环境,缺氧条件下WPMY-1细胞膜表面各类生长因子受体表达均增加,以EGFR为例,缺氧培养4 h后EGFR mRNA为0.34±0.04,略高于有氧条件下的0.29±0.01,但两者比较差异无统计学意义(P=0.149);此后观察时间点缺氧条件下细胞EGFR mRNA表达均显著高于有氧条件下。免疫组化显示缺氧条件培养4 h后细胞膜表面EGFR阳性表达为(7.11±0.31)/200个细胞,显著高于有氧条件的(6.00±0.29)/200个细胞(P=0.011),此后观察时间点除12 h差异无统计学意义外(P=0.166),缺氧条件下细胞EGFR蛋白表达均显著高于有氧条件下。结论缺氧可上调前列腺间质细胞膜表面上述各类生长因子受体表达。
Objective To observe the different expression of somatomedin-receptor in cell membrane of prostate stromal cell at anoxic or normoxic condition.Methods Prostate stromal cell line WPMY-1 was cultured in vitro.At 4,8,12,24,48 h after cell had been seeded,the gene and protein expression of epidermal growth factor receptor（EGFR）,fibroblast growth factor receptor（FGFR）,transforming growth factor receptor（TGFR）,insulin-like growth factor receptor（IGFR） and vascular endothelial growth factor receptor（VEGFR） in prostate stromal cell were tested by RT-PCR and immunohistochemistry methods,respectively.Results The expression of EGFR/FGFR/TGFR/IGFR/VEGFR significantly increased in anoxic prostate stromal cell as compared to normoxic prostate stromal cell.EGFR mRNA was 0.34±0.04 in anoxic prostate stromal cell tested by RT-PCR and was higher than that in normoxic prostate stromal cell（0.29±0.01）,but there was no significant difference between them（P=0.149）.At 8h,EGFR mRNA was 0.47±0.02 in anoxic prostate stromal cell,and was significantly higher than 0.37±0.01 in normoxic prostate stromal cell（P=0.01）;At 12,24,48 h,EGFR mRNA in anoxic prostate stromal cell were all significantly higher than those in normoxic prostate stromal cell.There were 7.11±0.31 EGFR positive expression cells per 200 cells in anoxic condition at 4 h after seeded by immunohistochemistry,significantly higher than 6.00±0.29 in normoxic condition（P=0.011）.Except for 12 h（P=0.166）,the difference between in anoxic condition and normoxic condition was significant（all P0.05）.Conclusion Anoxic prostate stromal cell can upregulate the expression of somatomedin-receptor.
出处
《中华全科医学》
2012年第5期667-668,707,F0003,共4页
Chinese Journal of General Practice
基金
广东省自然科学基金重点项目(8251001002000001)
关键词
缺氧
生长调节素
前列腺间质细胞
细胞培养
Hypoxia conditions
Somatomedin
Prostate stromal cell
Cell culture
