淫羊藿苷对前列腺癌原位移植瘤模型小鼠雄激素受体信号通路的影响
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摘要
目的:探讨淫羊藿苷对前列腺癌原位移植瘤模型SCID小鼠雄激素受体信号通路的影响机制。方法:雄性SCID小鼠64只,均采用前列腺腺体背外侧包膜内注射人前列腺癌细胞株(LNCaP)悬液的方法构建前列腺癌原位移植瘤模型,然后分为移植瘤组、10 mg·kg~(-1)组、40 mg·kg~(-1)组和80 mg·kg~(-1)组,除移植瘤组灌生理盐水对照外其它3组灌胃给予相应剂量的淫羊藿苷治疗5周。采用western blotting检测前列腺癌特异性抗原(PSA)和磷酸化AR(p-AR)表达,采用RT-PCR检测治疗前后前列腺瘤体雄激素受体(AR)和张力蛋白同源第10号染色体缺失的磷酸酶基因(PTEN)表达,采用流式细胞学方法检测前列腺体肿瘤瘤体LNCaP前列腺癌细胞在肿瘤瘤体增殖周期。结果:40 mg·kg~(-1)组和80 mg·kg~(-1)组治疗后AR mRNA为(0.25±0.02,0.27±0.03)、pAR为(1.45±0.22,1.64±0.24),PSA为(0.31±0.02,0.38±0.05),两组PSA、p-AR和AR mRN治疗后相对表达量均低表达,而PTEN mRNA高表达(0.91±0.07,0.95±0.09),与治疗前和移植瘤组比较差异均有统计学意义(P <0.05)。两组治疗后抑瘤率分别为(41.59±4.51)%和(42.76±5.13)%,瘤质量为(86.34±9.07,84.73±7.58)mg、瘤体积为(11.83±0. 84,10.27±1.14)mm2,均较同组治疗前和移植瘤组比较明显降低(P <0.05);两组治疗后G0/G1期比例降低[两组G0/G1分别为(34.97±4.52,35.03±3.97)%、且S期比例明显提高[两组S期比例分别为(39.59±5.03,40.27±4.82)%],与同组治疗前和移植瘤组比较差异均均有统计学意义(P <0.05)。结论:淫羊藿苷抑制LNCaP增殖的机制应与其抑制雄激素受体信号通路中AR的磷酸化并增强PTEN表达而将癌细胞增阻滞于S期有关。
Objective: To investigate the effects of icariin on androgen receptor signaling pathway in SCID mice model of orthotopic prostate cancer transplantation. Methods: Sixty-four male SCID mice were injected with human LNCaP prostate cancer cell suspension into the dorsolateral capsule of prostate gland to establish orthotopic prostate cancer transplantation model. The mice were divided into transplantation tumor group, 10 mg·kg~(-1) group, 40 mg·kg~(-1) group and 80 mg·kg~(-1) group. Intragastric administration of icariin was given for 5 weeks. The expression of androgen receptor(AR)and tensin homologous phosphatase gene(PTEN) deleted on chromosome 10 was detected by RT-PCR. Western blotting was used to detect the specificity of prostate cancer antigen(Prostate specific antigen, PSA) and phosphorylated AR(pAR), method of flow cytometry was used for the detection of LNCaP prostate cancer cell cycle. Results: After treatment,AR mRNA of 40 mg·kg~(-1) group and 80 mg·kg~(-1) group(0.25 ± 0.02, 0.27 ± 0.03), p-AR(1.45 ± 0.22, 1.64 ± 0.24), PSA(0.31 ± 0.02, 0.38 ± 0.05), and relative expression of PSA, p-AR and AR mRN were all low, while PTEN mRNA was high(0.91 ± 0.07, 0.95 ± 0.09). The difference between the two groups was statistically significant(P < 0.05). There was significant difference between pre transplant group and transplanted tumor group(P < 0.05). After treatment, the tumor inhibition rates were(41.59 ± 4.51)% and(42.76 ± 5.13)%, the tumor mass was(86.34 ± 9.07, 84.73 ± 7.58) mg, and the tumor volume was(11.83 ± 0.84, 10.27 ± 1.14) mm2, All of them were significantly lower than those of the same group before treatment and the xenograft group(P < 0.05). After treatment, the proportion of G0/G1 phase decreased in both groups [34.97 ± 4.52, 35.03 ± 3.97]%, and the proportion of S phase increased significantly [39.59 ± 5.03, 40.27 ±4.82%], and the difference was statistically significant before treatment and in the transplanted tumor group(P < 0.05).Conclusion: The mechanism of Icariin inhibiting the proliferation of LNCaP cells is related to its inhibition of AR phosphorylation in androgen receptor signaling pathway and enhancement of PTEN expression.
Objective: To investigate the effects of icariin on androgen receptor signaling pathway in SCID mice model of orthotopic prostate cancer transplantation. Methods: Sixty-four male SCID mice were injected with human LNCaP prostate cancer cell suspension into the dorsolateral capsule of prostate gland to establish orthotopic prostate cancer transplantation model. The mice were divided into transplantation tumor group, 10 mg·kg~(-1) group, 40 mg·kg~(-1) group and 80 mg·kg~(-1) group. Intragastric administration of icariin was given for 5 weeks. The expression of androgen receptor(AR)and tensin homologous phosphatase gene(PTEN) deleted on chromosome 10 was detected by RT-PCR. Western blotting was used to detect the specificity of prostate cancer antigen(Prostate specific antigen, PSA) and phosphorylated AR(pAR), method of flow cytometry was used for the detection of LNCaP prostate cancer cell cycle. Results: After treatment,AR mRNA of 40 mg·kg~(-1) group and 80 mg·kg~(-1) group(0.25 ± 0.02, 0.27 ± 0.03), p-AR(1.45 ± 0.22, 1.64 ± 0.24), PSA(0.31 ± 0.02, 0.38 ± 0.05), and relative expression of PSA, p-AR and AR mRN were all low, while PTEN mRNA was high(0.91 ± 0.07, 0.95 ± 0.09). The difference between the two groups was statistically significant(P < 0.05). There was significant difference between pre transplant group and transplanted tumor group(P < 0.05). After treatment, the tumor inhibition rates were(41.59 ± 4.51)% and(42.76 ± 5.13)%, the tumor mass was(86.34 ± 9.07, 84.73 ± 7.58) mg, and the tumor volume was(11.83 ± 0.84, 10.27 ± 1.14) mm2, All of them were significantly lower than those of the same group before treatment and the xenograft group(P < 0.05). After treatment, the proportion of G0/G1 phase decreased in both groups [34.97 ± 4.52, 35.03 ± 3.97]%, and the proportion of S phase increased significantly [39.59 ± 5.03, 40.27 ±4.82%], and the difference was statistically significant before treatment and in the transplanted tumor group(P < 0.05).Conclusion: The mechanism of Icariin inhibiting the proliferation of LNCaP cells is related to its inhibition of AR phosphorylation in androgen receptor signaling pathway and enhancement of PTEN expression.
引文
1韩苏军,张思维,陈万青,等.中国前列腺癌发病现状和流行趋势分析.临床肿瘤学杂志,2013,18(4):330-334.
2刘璐,徐莉春.雄激素受体信号通路介导抗雄激素效应.中国公共卫生,2010,26(12):1598-1599.
3 Kim W,Ryan C J.Androgen receptor directed therapies in castration resistant metastatic prostate cancer.Curr Treat Option On,2012,13(2):189-200.
4孙健玮,王剑松,刘子超,等.前列腺癌PTEN蛋白与雄激素受体的表达.中国医科大学学报,2014,43(4):316-319.
5 Yoshimoto M,Ding K,Sweet J M,et al.PTEN losses exhibit heterogeneity in multifocal prostatic adenocarcinoma and are associated with higher Gleason grade.Mod Pathol,2013,26(3):435-447.
6奚春.多种血清肿瘤标志物联合前列腺特异度抗原检测在前列腺癌诊断中的价值分析.中国卫生检验杂志,2017,14(24):3585-3587.
7张温花,张文超,于远东,等.淫羊藿苷对前列腺癌细胞株活力、迁移与侵袭的作用.中国病理生理杂志,2017,33(6):1017-1020.
8饶红,武福云,陈德森,等.淫羊藿苷对BALB/c-nu裸鼠前列腺癌组织磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路及E-钙黏蛋白的影响.中国中医急症,2018,27(5):789-792,796.
9王金秋,李志,何丹,等.SCID小鼠原位接种LNCaP前列腺癌模型的建立术.北京农业职业学院学报,2012,26(5):19-22.
10张洪涛,张静宇,张炳团,等.BC369对鼠源肾癌(REN-ca)抑瘤效应的研究.实用癌症杂志,2014,29(9):1053-1054.
11方芳,赵臣,郝峰,等.CD147对前列腺癌细胞雄激素受体磷酸化的影响及其作用机制.山东医药,2017,57(28):9-11.
12翟葳,冯极灵,席志超,等.Guttiferone K诱导人前列腺癌LNCaP细胞G_(0/1)期阻滞的作用研究.世界科学技术-中医药现代化,2017,19(2):235-240.
13胡蕾蕾,段小群,卢曦,等.淫羊藿素对雌激素依赖性乳腺癌MCF-7细胞作用的影响.中国实验方剂学杂志,2012,18(14):155-158.
14中国科学院中国植物志编辑委员会.淫羊藿M:中国植物志第29卷.北京:科学出版社,2004:296-298.
15赖丽娟,谢佳丽,黄志华,等.淫羊藿素的抗肿瘤作用及机制研究进展.中药药理与临床,2016,12(6):235-238.
16陈鹏,刘文和,颜林淋,等.淫羊藿黄酮对去势大鼠骨生物力学性能的影响.世界科学技术-中医药现代化,2014,16(8):1814-1818.
17曹爱娇,邵瑞,王彧,等.中药通过调节雄激素受体信号通路治疗前列腺癌的研究进展.中国临床药理学杂志,2017,33(13):1263-1266.
18陈少秀,饶红,陈德森,等.淫羊藿苷对BALB/c-nu裸小鼠雄激素依赖性前列腺癌雄激素受体信号转导通路的影响.长春中医药大学学报,2018,34(3):436-438.
19王应熊.肿瘤与遗传.分子与细胞,北京,人民卫生出版社,2016:23-33.
20詹启敏,陈杰.细胞周期与肿瘤转化医学.中国肿瘤临床,2014,41(1):1-7.
21 Dehm S M,Tindall D J.Molecular regulation of androgen action in prostate cancer.J Cell Biochem,2006,99(2):333-344.
22蒋宏毅,赵晓昆,钟朝晖,等.PTEN/MMAC1/TEP1、TGF-β1在前列腺癌及前列腺增生中的表达及其意义.中国现代医学杂志,2008,18(9):1221-1225.
23孙健玮,王剑松.PTEN基因与前列腺癌.昆明医科大学学报,2013,12(11):139-142.
24饶红,武福云,何华琼,等.淫羊藿苷对前列腺癌原位移植瘤SCID小鼠脂肪酸合成酶和LNCaP细胞生物学行为的影响.实用药物与临床,2018,21(5):499-501.
2刘璐,徐莉春.雄激素受体信号通路介导抗雄激素效应.中国公共卫生,2010,26(12):1598-1599.
3 Kim W,Ryan C J.Androgen receptor directed therapies in castration resistant metastatic prostate cancer.Curr Treat Option On,2012,13(2):189-200.
4孙健玮,王剑松,刘子超,等.前列腺癌PTEN蛋白与雄激素受体的表达.中国医科大学学报,2014,43(4):316-319.
5 Yoshimoto M,Ding K,Sweet J M,et al.PTEN losses exhibit heterogeneity in multifocal prostatic adenocarcinoma and are associated with higher Gleason grade.Mod Pathol,2013,26(3):435-447.
6奚春.多种血清肿瘤标志物联合前列腺特异度抗原检测在前列腺癌诊断中的价值分析.中国卫生检验杂志,2017,14(24):3585-3587.
7张温花,张文超,于远东,等.淫羊藿苷对前列腺癌细胞株活力、迁移与侵袭的作用.中国病理生理杂志,2017,33(6):1017-1020.
8饶红,武福云,陈德森,等.淫羊藿苷对BALB/c-nu裸鼠前列腺癌组织磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路及E-钙黏蛋白的影响.中国中医急症,2018,27(5):789-792,796.
9王金秋,李志,何丹,等.SCID小鼠原位接种LNCaP前列腺癌模型的建立术.北京农业职业学院学报,2012,26(5):19-22.
10张洪涛,张静宇,张炳团,等.BC369对鼠源肾癌(REN-ca)抑瘤效应的研究.实用癌症杂志,2014,29(9):1053-1054.
11方芳,赵臣,郝峰,等.CD147对前列腺癌细胞雄激素受体磷酸化的影响及其作用机制.山东医药,2017,57(28):9-11.
12翟葳,冯极灵,席志超,等.Guttiferone K诱导人前列腺癌LNCaP细胞G_(0/1)期阻滞的作用研究.世界科学技术-中医药现代化,2017,19(2):235-240.
13胡蕾蕾,段小群,卢曦,等.淫羊藿素对雌激素依赖性乳腺癌MCF-7细胞作用的影响.中国实验方剂学杂志,2012,18(14):155-158.
14中国科学院中国植物志编辑委员会.淫羊藿M:中国植物志第29卷.北京:科学出版社,2004:296-298.
15赖丽娟,谢佳丽,黄志华,等.淫羊藿素的抗肿瘤作用及机制研究进展.中药药理与临床,2016,12(6):235-238.
16陈鹏,刘文和,颜林淋,等.淫羊藿黄酮对去势大鼠骨生物力学性能的影响.世界科学技术-中医药现代化,2014,16(8):1814-1818.
17曹爱娇,邵瑞,王彧,等.中药通过调节雄激素受体信号通路治疗前列腺癌的研究进展.中国临床药理学杂志,2017,33(13):1263-1266.
18陈少秀,饶红,陈德森,等.淫羊藿苷对BALB/c-nu裸小鼠雄激素依赖性前列腺癌雄激素受体信号转导通路的影响.长春中医药大学学报,2018,34(3):436-438.
19王应熊.肿瘤与遗传.分子与细胞,北京,人民卫生出版社,2016:23-33.
20詹启敏,陈杰.细胞周期与肿瘤转化医学.中国肿瘤临床,2014,41(1):1-7.
21 Dehm S M,Tindall D J.Molecular regulation of androgen action in prostate cancer.J Cell Biochem,2006,99(2):333-344.
22蒋宏毅,赵晓昆,钟朝晖,等.PTEN/MMAC1/TEP1、TGF-β1在前列腺癌及前列腺增生中的表达及其意义.中国现代医学杂志,2008,18(9):1221-1225.
23孙健玮,王剑松.PTEN基因与前列腺癌.昆明医科大学学报,2013,12(11):139-142.
24饶红,武福云,何华琼,等.淫羊藿苷对前列腺癌原位移植瘤SCID小鼠脂肪酸合成酶和LNCaP细胞生物学行为的影响.实用药物与临床,2018,21(5):499-501.