He Guifang, Mu Tianlong, Yuan Yali, Yang Wenyan, Zhang Yuan, Chen Qingyun, Bian Meilu, Pan Yanshu, Xiang Qing, Chen Zhihua, Sun Aiping
Department of Gynecology & Obstetrics, China-Japan Friendship Hospital, Beijing, China, 100029.
Department of Pathology, Oriental Hospital of Beijing University of Chinese Medicine, Beijing, China, 100078.
J Cancer. 2019 Jul 10;10(17):4114-4122. doi: 10.7150/jca.30690. eCollection 2019.
Curcumin, as a high effect and low toxicity anti-cancer drug and photosensitiser, has synergistic and complementary effects with photodynamic therapy (PDT). However, due to its unclear mechanism, PDT's application and efficacy were limited. Notch signaling pathway, which is highly correlates with carcinogenesis and development of cervical cancer, could be a potential therapeutic targets to improve the effectiveness of PDT. Therefore, in this study, we explored the effects of Notch signaling pathway in cervical cancer by curcumin mediated PDT with/without Notch receptor blocker (DAPT), and hope to elucidate its mechanism. Firstly, the effect on the proliferation of cervical cancer Me180 cells were detected with MTT assay, and apoptosis were detected with Annexin V-FITC/PI combined with flow cytometry. Secondly, after establishment of nude mice model, dividing the experimental animals into model group, curcumin PDT group, simple DAPT group, and curcumin-PDT+DAPT group, and analyzing tumor volume changes as well as HE staining in each group. mRNA and protein expression of gene Notch-1 and its downstream NF-κB and VEGF were observed with RT-PCR, immunohistochemical staining and Western-blot with/without inhibition of Notch signaling pathway by DAPT, both in vivo and in vitro experiments. We found both DAPT and curcumin-PDT can inhibit the proliferation and induce apoptosis of cervical cancer cell. The two have synergistic effect in vitro and in vivo. This effect can effectively block the conduction of Notch signaling pathway, which is associated with down-regulation of the expression of Notch1 and NF-κB. Notch signaling pathway could be one of the targets of curcumin-PDT photodynamic therapy.
姜黄素作为一种高效低毒的抗癌药物和光敏剂，与光动力疗法（PDT）具有协同和互补作用。然而，由于其作用机制尚不清楚，PDT的应用和疗效受到限制。Notch信号通路与宫颈癌的发生发展高度相关，可能是提高PDT疗效的潜在治疗靶点。因此，在本研究中，我们通过姜黄素介导的PDT联合或不联合Notch受体阻滞剂（DAPT），探讨了Notch信号通路在宫颈癌中的作用，希望阐明其机制。首先，采用MTT法检测对宫颈癌Me180细胞增殖的影响，采用Annexin V-FITC/PI结合流式细胞术检测细胞凋亡。其次，建立裸鼠模型后，将实验动物分为模型组、姜黄素PDT组、单纯DAPT组和姜黄素-PDT+DAPT组，分析各组肿瘤体积变化及HE染色情况。在体内外实验中，通过DAPT抑制Notch信号通路，采用RT-PCR、免疫组织化学染色和Western blot观察基因Notch-1及其下游NF-κB和VEGF的mRNA和蛋白表达。我们发现DAPT和姜黄素-PDT均能抑制宫颈癌细胞的增殖并诱导其凋亡。两者在体内外均具有协同作用。这种作用可有效阻断Notch信号通路的传导，这与Notch1和NF-κB表达的下调有关。Notch信号通路可能是姜黄素-PDT光动力治疗的靶点之一。
