甲硫哒嗪联合孕激素抑制子宫内膜癌细胞增殖的作用机制

Mechanism of thioridazine plus medroxyprogesterone in the treatment of endometrial cancer

目的明确甲硫哒嗪（THIO）联合醋酸甲羟孕酮（MPA）在体外对子宫内膜癌细胞Ishikawa（ISK）及KLE增殖与凋亡的影响及其作用机制。方法通过CCK-8法观察THIO联合MPA对ISK和KLE增殖的抑制作用，选择药物作用的最佳浓度及时间；实时定量PCR和Western印迹方法检测用药前后PRB、DRD2以及P13K／AKT信号通路中p-AKT／AKT的变化；流式细胞术比较THIO联合MPA与单一MPA对细胞凋亡的差异。结果与单一MPA相比，THIO联合MPA明显抑制ISK（52．5％±2．6％比37．3％±0．3％，P〈0．05）和KLE（97．7％±0．7％比50．0％±0．4％，P〈0．01）的增殖；促进ISK（34．O％±1．4％比50．5％±2．4％，P〈0．01）和KLE（5．5％±3．6％比11．3％±0．7％，P〈0，01）的凋亡；显著提高ISK（5．4±0．5比17．8±3．7，P〈0．05）和KLE（0．5±0．1比7．9±1．6，P〈0．05）的PRB表达、DRD2表达（1．8±0．2比7．5±0．3，P〈0．05；1．4±0．1比11．4±2．1，P〈0．01）；同时降低P13K／AKT信号通路中p-AKT／AKT水平。结论THIO显著增强MPA对子宫内膜癌细胞的抑制增殖及促进凋亡作用，其作用可能与PRB和DRD2介导的P13K／AKT信号通路相关。

Objective To explore the in vitro effects of thioridazine （THIO） plus medroxyprogesterone （MPA） on the proliferation and apoptosis of endometrial cancer cell lines （Ishikawa ＆ KLE） and examine the mechanism in the treatment of endometrial cancer. Methods CCK-8 assay was employed for detecting the influence of THIO plus MPA on the proliferation and apoptosis of endometrial cancer cells （ ISK ＆ KLE）. And the concentration and timepoints of drugs were determined according to the results. Real-time polymerase chain reaction （PCR） and Western blot were used to detect the expression levels of PRB, DRD2 and p-AKT/AKT in PI3K/AKT signal pathway. Flow cytometry was applied for detecting the apoptosis of combination （THIO ＋ MPA） and MPA groups. Results Compared to MPA group, the proliferation inhibiting effect of combination group increased significantly in ISK and KLE cells （52. 5 % ±2.6% vs37.3% ＋0.3%, P〈0.05; 97.7% ±0.7% vs50.0% ±0.4%, P〈0.001）; the apoptotic rates increased （34.0% ±1.4% vs 50.5% ±2.4%, P〈0.01; 5.5% ＋3.6% vs 11.3% ±0.7%, P〈 0. 01 ） ; the expression levels of PRB and DRD2 were up-regulated （ all P 〈 0. 05 ）. And the ratio of p-AKT/ AKT decreased obviously. Conclusion Thioridazine significantly enhances the effects of MPA on proliferative inhibition and apoptotic promotion in endometrial cancer cells. And it may be associated with the PRB/DRD2-mediated PI3K/AKT signal pathway.