目的 研究百里醌对卵巢癌生长的影响及其作用机制。方法 百里醌作用卵巢癌细胞株SKOV3后,CCK-8法检测细胞增殖;ELISA法检测细胞凋亡;荧光探针法检测细胞内活性氧(ROS)水平;Western blotting检测胞核蛋白Nrf2、胞浆蛋白Keap1、Akt、p-Akt、NQO1、GCLC的表达;实时荧光定量PCR(RT-qPCR)检测细胞中NQO1和GCLC mRNA水平。制备裸鼠卵巢癌皮下移植瘤模型,观察百里醌对肿瘤生长的影响;免疫组化检测肿瘤组织Nrf2、NQO1和GCLC的阳性表达。结果 与对照组比较,百里醌明显抑制SKOV3细胞生长(P<0.05、0.01),并诱导细胞凋亡(P<0.01);升高细胞内ROS水平(P<0.05);下调胞核Nrf2蛋白及胞浆p-Akt蛋白表达(P<0.05),上调Keap1蛋白表达(P<0.001);下调NQO1、GCLC的mRNA和蛋白表达(P<0.05、0.01)。百里醌抑制裸鼠卵巢癌皮下移植瘤的生长(P<0.01),降低肿瘤组织中Nrf2、NQO1、GCLC的阳性表达(P<0.05、0.01)。结论 百里醌抑制卵巢癌生长,其机制可能是抑制胞核Nrf2蛋白表达,促进癌细胞内ROS的产生,诱导细胞凋亡。

Objective To study the effect and mechanism of thymoquinone in the growth inhibition of ovarian cancer. Methods After ovarian cancer cells SKOV3 were treated with thymoquinone, cell growth was observed by the CCK-8 assay and apoptosis was evaluated by ELISA. ROS in SKOV3 cells was detected by fluorescent probe. The expression of Keap1 protein, nuclear Nrf2 protein, P-Akt protein, Akt protein, NQO1 protein, and GCLC protein in SKOV3 cells was analyzed by Western blotting assay. NQO1 and GCLC mRNA were analyzed by RT-PCR. SKOV3 cells were injected into nude mice to establish engrafted tumor model. Immunohistochemistry was used to detect the positive expression of NQO1, GCLC, and Nrf2 in the ovarian tumors. Results Compared with the control group, thymoquinone significantly inhibited the proliferation of SKOV3 cells and increased the apoptosis (P < 0.05, 0.01). Thymoquinone could lessen the expression of nuclear Nrf2 protein and P-Akt protein (P < 0.05), but significantly increase the expression of Keap1 protein (P < 0.001). Protein and mRNA expression of NQO1 and GCLC in SKOV3 cells treaded by thymoquinone, were lower than those in the control group (P < 0.05, 0.01). The positive expression of NQO1, GCLC, and Nrf2 was also decreased in ovarian tumors after treatment of thymoquinone (P < 0.05, 0.01). Conclusion Thymoquinone could inhibit the proliferation of ovarian cancer cells by inhibiting the expression of nuclear Nrf2, increasing the generation of ROS in ovarian cancer cells to promote apoptosis.

国家自然科学基金资助项目(81070510)