Alpha-2A肾上腺素能受体在去势抵抗性前列腺癌患者外周血的表达及临床意义

Expression and clinical significance of alpha-2a adrenergic receptor in peripheral blood of patients with castration-resistant prostate cancer

目的探讨Alpha-2A肾上腺素能受体(ADRA2A)在去势抵抗性前列腺癌患者外周血的表达及临床意义。方法采用蛋白印迹法和实时定量反转录聚合酶链反应, 检测经长期恩杂鲁胺(ENZ)药物处理的耐药人前列腺癌细胞LNCaP-ENZ和本身抵抗ENZ的人前列腺癌细胞22Rv1的ADRA2A表达量变化;瞬时转染ADRA2A后测定上述耐药细胞存活率;采用酶联免疫吸附试(ELISA)验测定来源于江南大学附属医院的20例正常人(N)、20例前列腺增生(BPH)、30例原发性前列腺癌(LPCa)及10例去势抵抗性前列腺癌(CRPC)患者的外周血中ADRA2A含量变化;GEPIA数据库中针对PCa患者数据分析ADRA2A对生存能力的影响。多样本比较采用单因素方差分析, 两样本比较采用t检验。结果本研究结果显示在mRNA水平, 耐药ENZ的LNCaP-ENZ细胞ADRA2A表达量(14.44±0.629)高于正常前列腺癌LNCaP细胞(1±0), 差异有统计学意义(t=21.370, P<0.05);在蛋白水平, LNCaP-ENZ细胞ADRA2A表达量(4.243±0.217)高于正常前列腺癌LNCaP细胞(1±0), 差异有统计学意义(t=14.970, P<0.05);LNCaP-ENZ细胞中敲低ADRA2A, si-ADRA2A组细胞存活率[(60.650±2.451)%]明显低于对照组[(100±0)%]及si-neg组[(100.300±0.633)%], 差异有统计学意义(F=243.800, P<0.05);ELISA结果显示LPCa组(3.084±0.196)和CRPC组表达量(4.364±0.306)均显著高于BPH组(2.203±0.202)和N组(2.192±0.227), 差异有统计学意义(F=13.640, P<0.05), CRPC组比较LPCa组, ADRA2A表达量显著升高, 差异有统计学意义(P<0.05);GEPIA分析发现在PCa中, ADRA2A高表达使患者生存率降低, 但其差异无统计学意义(P>0.05)。结论 ADRA2A在CRPC患者中表达量最高, 有助于CRPC患者诊断和监测。

Objective To investigate the expression and clinical significance of alpha-2a adrenergic receptor(ADRA2A)in peripheral blood of patients with castration-resistant prostate cancer.Methods Western blotting and real-time quantitative reverse transcription polymerase chain reaction were used to detect the expression of ADRA2A in long-term enzalutamide(ENZ)treated resistant human prostate cancer cells LNCaP-ENZ and ENZ self-resistant human prostate cancer cells 22Rv1.The survival rate of these drug-resistant cells was measured after transient transfection of ADRA2A.The enzyme linked immunosorbent assay(ELISA)was used to detect the ADRA2A content changes in 20 cases of normal controls(N),20 cases of benign prostatic hyperplasia(BPH),30 cases of localized prostate cancer(LPCa)and 10 cases of castration-resistant prostate cancer(CRPC)patients in peripheral blood collected from the affiliated hospital of Jiangnan University.The impact of ADRA2A on survival ability of PCa patients in GEPIA database was analyzed.One-way analysis of variance was used for comparison of multi-samples,and T-test was used for comparison of two samples.Results This study found that as compared with LNCaP cells(1±0),the expression of ADRA2A mRNA in ENZ resistant cells(LNCaP-ENZ)(14.44±0.629)was increased and the difference was statistically significant(t=21.370,P<0.05).As compared with LNCaP cells(1±0),the expression of ADRA2A protein in LNCaP-ENZ(4.243±0.217)was increased and the difference was statistically significant(t=14.970,P<0.05).When ADRA2A was knocked down in LNCaP-ENZ,the cell survival rate in si-ADRA2A group[(60.650±2.451)%]was significantly lower than that in the control group[(100±0)%]and si-neg group[(100.300±0.633)%],and the difference was statistically significant(F=243.800,P<0.05),but there was no significant difference between the control group and si-neg group(P>0.05).The ELISA results showed that the expression of ADRA2A in LPCa group(3.084±0.196)and CRPC group(4.364±0.306)was significantly higher than that in BPH group(2.203±0.202)and normal control group(2.192±0.227),and the difference was statistically significant(F=13.640,P<0.05).As compared with LPCa group,the expression of ADRA2A in CRPC group was significantly increased,and the difference was statistically significant(P<0.05).GEPIA analysis showed that the high expression of ADRA2A reduced the survival rate of PCa patients,but the difference was no statistically significant(P>0.05).Conclusion ADRA2A has the highest expression level in CRPC patients,which is expected to become a new biomarker for the diagnosis and monitoring of CRPC patients.