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目的探讨血红素加氧酶-1(HO-1)在人前列腺癌组织中的表达,以及HO-1活性对体外培养的前列腺癌细胞(PC3)的影响。方法 收集我院2000年1月至2007年4月前列腺癌根治性手术标本30例,前列腺增生开放手术大体标本30例。行RT-PCR检测HO-1的mRNA表达,免疫组化检测HO-1的表达。应用ZnPP、CoPP干预体外培养的PC3细胞,24h后,RT-PCR检测HO-1 mRNA的表达,MTT法酶标检测仪检测细胞A570nm值,流式细胞术检测细胞凋亡。结果人前列腺癌组织HO-1的mRNA表达明显高于良性增生前列腺组织;免疫组化前列腺癌中HO-1表达阳性率为76.9%,良性前列腺组织仅少许有微弱表达。应用ZnPP、CoPP干预PC3细胞后,RT-PCR可见ZnPP抑制PC3细胞HO-1表达,且随浓度增加抑制作用增加;CoPP可诱导HO-1表达,随浓度增加表达增强。MTT检测显示,ZnPP组PC3细胞A值随浓度增加而减少,而CoPP组随浓度增加A值增加,呈现浓度依赖性(P<0.05)。流式细胞技术结果显示,ZnPP可诱导PC3细胞凋亡,CoPP可保护细胞减少凋亡。结论人前列腺癌组织中HO-1基因表达及蛋白水平均显著高于良性前列腺组织;体外细胞实验证实,ZnPP可抑制前列腺癌PC3细胞系HO-1活性,抑制其增殖,诱导其凋亡。而CoPP可诱导前列腺癌PC3细胞系HO-1表达,抑制其凋亡,诱导其增殖。
Objective To investigate the expression of heme oxygenase-1 (HO-1) in human prostate cancer and the effect of HO-1 activity on prostate cancer cells (PC3) cultured in vitro. Methods Thirty cases of radical prostatectomy specimens and 30 specimens of benign prostatic hyperplasia were collected from January 2000 to April 2007 in our hospital. The mRNA expression of HO-1 was detected by RT-PCR and the expression of HO-1 was detected by immunohistochemistry. The PC3 cells cultured in vitro were treated with ZnPP and CoPP, and the expression of HO-1 mRNA was detected by RT-PCR 24h after treatment. The A570nm was detected by MTT assay and the apoptosis was detected by flow cytometry. Results The mRNA expression of HO-1 in human prostate cancer tissue was significantly higher than that in benign prostatic hyperplasia. The positive rate of HO-1 in immunohistochemical prostate cancer was 76.9%, and only a little weak expression in benign prostatic tissue. After ZnPP and CoPP were used to intervene in PC3 cells, RT-PCR showed that ZnPP inhibited the expression of HO-1 in PC3 cells and increased with the increase of concentration. CoPP induced the expression of HO-1 and increased with the increase of concentration. MTT assay showed that the A value of PC3 cells in ZnPP group decreased with increasing concentration, while that of CoPP group increased in a concentration-dependent manner (P <0.05). Flow cytometry showed that ZnPP induced PC3 cell apoptosis and CoPP could protect cells from apoptosis. Conclusions HO-1 gene expression and protein level in human prostate cancer tissues were significantly higher than those in benign prostatic tissues. In vitro experiments confirmed that ZnPP could inhibit HO-1 activity, inhibit proliferation and induce apoptosis of PC-3 cells. However, CoPP can induce the expression of HO-1 in prostate cancer PC3 cell line, inhibit its apoptosis and induce its proliferation.