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目的:观察在缺氧条件下前列腺上皮细胞体外培养生长类激素表达的变化。方法:前列腺上皮细胞株RWPE-1体外培养。在缺氧或有氧培养环境中分别于接种后4、8、12、24、48 h检测其生长类激素[表皮生长因子(EGF),碱性成纤维细胞生长因子(bFGF),转化生长因子β(TGF-β),胰岛素样生长因子-1(IGF-1),血管内皮生长因子(VEGF)]基因和蛋白表达。基因表达采用RT-PCR法检测,蛋白表达采用ELISA法检测。结果:RT-PCR结果显示,随时间延长各类生长因子基因表达均上调,缺氧条件较有氧条件上调尤为明显。以bFGF mRNA表达为例,接种后8 h缺氧条件和有氧条件下bFGF mRNA分别为0.14±0.01和0.12±0.01,差异有统计学意义(P=0.01),至48 h时分别为0.29±0.01和0.14±0.01(P<0.001)。ELISA结果显示,上皮细胞TGF-β表达在缺氧条件下亦显著增加,接种后4 h缺氧和有氧条件下分别为0.32±0.01和0.26±0.01,差异有统计学意义(P=0.017),至48 h时分别为1.56±0.13和0.87±0.06(P<0.001)。其他4种生长因子的蛋白表达在有氧和无氧条件下没有显著性差异。结论:缺氧可刺激前列腺上皮细胞上述各类生长因子基因表达的上调,但仅TGF-β分泌增加。
OBJECTIVE: To observe the changes of the growth hormone secreted by prostate epithelial cells cultured in vitro under hypoxic conditions. Methods: Prostate epithelial cell line RWPE-1 was cultured in vitro. The growth hormone (EGF, bFGF, transforming growth factor β (TGF-β), insulin-like growth factor-1 (IGF-1) and vascular endothelial growth factor (VEGF) The gene expression was detected by RT-PCR and the protein expression was detected by ELISA. Results: The results of RT-PCR showed that the expression of all kinds of growth factors were up-regulated over time, especially in hypoxia. Taking bFGF mRNA expression as an example, the bFGF mRNA was 0.14 ± 0.01 and 0.12 ± 0.01 respectively at 8 h after inoculation (P = 0.01) and 0.29 ± 48 h after 48 h 0.01 and 0.14 ± 0.01 (P <0.001). The results of ELISA showed that the expression of TGF-βin epithelial cells was also significantly increased under hypoxia conditions. The hypoxia and aerobic conditions at 4 h after inoculation were 0.32 ± 0.01 and 0.26 ± 0.01, respectively, with significant difference (P = 0.017) , 1.56 ± 0.13 and 0.87 ± 0.06, respectively, at 48 h (P <0.001). The other four kinds of growth factor protein expression under aerobic and anaerobic conditions there was no significant difference. CONCLUSION: Hypoxia can stimulate the up-regulation of the above-mentioned growth factor gene expression in prostate epithelial cells, but only increase the secretion of TGF-β.