有机硒通过下调AR及PSA抑制激素非依赖性前列腺癌细胞生长  

Organic selenium inhibits growth of hormone-independent prostate cancer cells by down-regulating AR and PSA

作　　者：杨丽娟[1,2] 葛贺[1] 安丽萍[3] 孙新 郭鹏 刘杰[1,2] 林珈羽[1] 刘艳波 YANG Li-juan;GE He;AN Li-ping;SUN Xin;GUO Peng;LIU Jie;LIN Jia-yu;LIU Yan-bo(Department of Pathophysiology,Medical College,School of Pharmacy,Beihua University,Jilin 132013,China;Jilin Provincial Key Laboratory of Molecular Geriatric Medicine,School of Pharmacy,Beihua University,Jilin 132013,China;Department of Microbiology and Biochemical Pharmacy,School of Pharmacy,Beihua University,Jilin 132013,China;Department of Nephropathy and Rheumatology,The Affiliated Hospital of Beihua University,Jilin 132011,China)

机构地区：[1]北华大学医学院病理生理学教研室,吉林吉林132013 [2]北华大学吉林省分子老年医学重点实验室,吉林吉林132013 [3]北华大学药学院微生物与生化药学教研室,吉林吉林132013 [4]北华大学附属医院肾病风湿病科,吉林吉林132011

出　　处：《中国病理生理杂志》2019年第11期1957-1965,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.813002226);吉林省科技厅自然科学研究计划项目(No.20190802011ZG);吉林省教育厅科学研究计划项目(No.JJKH20180329KJ; No.JJKH20170050KJ);吉林省卫生计生委科学研究计划项目(No.2018J095; No.2017J085)

摘　　要：目的:探讨甲基化硒酸(MSA)对人前列腺癌LNCaP细胞生长的抑制作用和甲基硒代半胱氨酸(MSC)延缓裸鼠去势后激素非依赖性前列腺癌的效应及相关机制。方法:将体外培养的LNCaP细胞与不同浓度的MSA共培养,磺酰罗丹明B(SRB)法检测MSA对细胞生长的抑制作用,流式细胞术检测细胞周期和凋亡,半定量RT-PCR及Western blot法检测雄激素受体(AR)及前列腺特异性抗原(PSA)的表达,免疫荧光和细胞化学染色法检测信号转导及转录激活因子3(STAT3)的表达。用BALB/c-nu/nu雄性裸鼠构建LNCaP细胞源性前列腺癌皮下移植瘤模型,待移植瘤直径达5 mm时,动物手术去势。将动物分为对照(mock)组(给予生理盐水200μL)和MSC组(给予5μg/gMSC),灌胃治疗,连续给药16周,监测肿瘤生长情况,ELISA法检测PSA的分泌规律,RT-PCR及Western blot检测肿瘤组织AR、PSA和STAT3的表达,TUNEL法检测肿瘤细胞的凋亡情况。结果:与mock组比较,MSA呈时间及剂量依赖性抑制LNCaP细胞生长,其抑制率可达65.32%±12.11%。MSA促进癌细胞凋亡,最大凋亡率为52.80%±2.87%,同时下调AR、PSA及STAT3的表达(P<0.05)。去势后MSC组的肿瘤复发时间延后,肿瘤生长速度减慢,同时下调前列腺癌移植瘤AR、PSA及STAT3表达(P<0.05)。结论:MSA呈时间及剂量依赖性抑制人前列腺癌LNCaP细胞生长,MSC延缓激素非依赖性前列腺癌发生,其机制与下调AR、PSA及STAT3表达有关。AIM: To investigate the growth-inhibiting effect of organic selenium methylseleninic acid(MSA) on human prostate cancer LNCaP cells and the retarding effect of methylselenocysteine(MSC) on the occurrence of subcutaneous hormone-independence prostate cancer(HIPC) xenografts in nude mice after castration, and to elucidate the underlying mechanisms. METHODS: After the LNCaP cells were treated with different concentrations of MSA, the growth of the cells was determined by sulforhodamine B(SRB) assay, and cell cycle and apoptosis were analyzed by flow cytometry. The expression of androgen receptor(AR) and prostate-specific antigen(PSA) at mRNA and protein levels was determined by RT-PCR and Western blot. The expression of signal transducer and activator of transcription 3(STAT3) was detected by immunofluorescence and immunohistochemical staining. LNCaP cells were cultured and digested with trypsin, and then the cells were inoculated into the back of the nude mice subcutaneously. When the palpable tumors had developed and reached 5 mm in diameter, the mice were divided randomly into 2 groups, and castration operation was performed. The mice in MSC group were given MSC at 5 μg/g by intragastric administration for 16 weeks, while the mice in mock group were treated with saline. The growth of the tumors was monitored and ELISA was applied to evaluate serum PSA levels. The mice were sacrificed at the end of the 16 th week. The tumors were excised, and the expression of AR, PSA and STAT3 at mRNA and protein levels in the xenograft was determined by RT-PCR and Western blot. TUNEL staining was applied to detect apoptotic cells. RESULTS: Compared with mock group, MSA inhibited the growth of LNCaP cells in a time-and dose-dependent manner and the inhibitory rate reached 65.32%±12.11%. MSA promoted the apoptosis of LNCaP cells and the maximum rate of apoptosis was 52.80%±2.87%. MSA inhibited AR, PSA and STAT3 expression. Compared with mock group, the recurrence of HIPC was delayed. The growth of xenografts in the mice treated

关 键 词：甲基化硒酸 甲基硒代半胱氨酸 前列腺癌 雄激素受体 前列腺特异性抗原 

分 类 号：R363.2[医药卫生—病理学] R737.25[医药卫生—基础医学]