双炔失碳酯α、β单体体外抗前列腺癌作用比较

Comparison of the Different Effects between Two Epimerides of Anordrin α and β Monomer on Human Prostatic Cancer in Vitro

关键词:  双炔失碳酯  差相异构体  人前列腺癌细胞  激素依赖性细胞  非激素依赖性细胞

Key words:  anordrin  epimerides  human prostatic cancer  androgen-dependent cells  androgen-independent cells

摘要:

目的 比较双炔失碳酯(ANO)的两个差向异构体a、b-ANO体外抗前列腺癌作用的差别。方法 氧化铝柱层析法分离纯化a、b单体，薄层分析法定性检测所分离的产物；在常规培养液中，以4～32 mmol·L-1的a、b-ANO分别作用于人前列腺癌非激素依赖型细胞DU145、PC-3和激素依赖型细胞L1A、LNCaP、RV1，作用时间分别为3 d，5 d，SRB法测细胞生存率；在去除激素培养液中，以不同浓度的a、b-ANO分别作用于激素依赖型细胞，同时加入雄激素拟似剂(R1881)，SRB法测细胞生存率；以32 mmol·L-1的a、b-ANO分别作用于LNCaP、DU145细胞，相差显微镜观察细胞形态和数量的变化。结果 对于非激素依赖型和激素依赖型细胞，a-ANO作用均强于b-ANO，且二者作用差距具有时间和剂量依赖性；a-ANO对抗雄激素的促细胞生长作用强于b-ANO，具有时间和剂量依赖性；相差显微镜观察结果表明，a-ANO作用后，细胞形态及数量变化较之b-ANO更为显著。结论 a-ANO体外抗前列腺癌作用强于b-ANO。

Abstract:

OBJECTIVE To investigate the different effects between two epimerides of anordrin(ANO) α and β monomer on human prostatic cancer. METHODS The monomers of α-ANO and β-ANO were purified and separated by alumina column chromatography, and were detected qualitatively by thin-layer chromatography；androgen-dependent and androgen-independent human prostatic cancer cells (androgen-dependent cells L1A, LNCaP, CWR22-RV1 and androgen-independent cells DU145, PC-3 ) were cultured with different concentrations (8, 16, 24, 32, 48 μmol·L-1) of α , β-ANO for 3 and 5 d respectively, SRB staining assay was used to detect cells survival; in androgen-depleted medium，L1A, LNCaP and CWR22-RV1 cells were cultured with different concentrations of α, β-ANO in the presence or absence of R1881 (a kind of androgen stimulant) for 3 and 5 d respectively, and SRB staining assay was used to detect cells survival；contrast phase microscope was used to observe the change of cellular morphology after LNCaP, DU145 cells being exposed to 32 μmol·L-1 α, β-ANO respectively. RESULTS The inhibition effect of α-ANO on human prostatic carcinoma cells was stronger than that of β-ANO in time and dose dependent manner; both α-ANO and β-ANO could antagonize the proliferation effect of R1881 and the effect of α-ANO was more stronger than of β-ANO in time and dose dependent manner; observed under contrast phase microscope, cells morphology change more significant after treatment with α-ANO than those treatment with β-ANO. CONCLUSION The suppressive effect of α-ANO on prostatic cancer cells line is stronger than that of β-ANO.