Wu Xue, Liu Ping, Zhang Haijun, Li Yuan, Salmani Jumah Masoud Mohammad, Wang Fei, Yang Ke, Fu Rong, Chen Zhewei, Chen Baoan
Department of Hematology and Oncology (Key Department of Jiangsu Medicine), Medical School, the Affiliated Zhongda Hospital, Southeast University, Nanjing, 210009, China.
Department of Gastroenterology, Medical School, The Second Hospital of Nanjing Affiliated to Southeast University, Nanjing, China.
BMC Cancer. 2017 Feb 21;17(1):147. doi: 10.1186/s12885-017-3145-4.
Wogonin is an encouraging choice for clinical use owing to its potent anti-tumor and anti-inflammatory effects with the high safety profile. However, wogonin for targeted therapy of lymphoma was not well addressed. In this study, we focused on its anticancer effect alongside with the underlying mechanisms for targeted therapy in EBV-positive lymphoma. This will facilitate its introduction to clinical use, which is planned in the near future.
Cell proliferation was studied by CCK8. Flow cytometry was used to analyze the apoptosis and the cycle arrest of cells. Further, we also used immunofluorescent staining to detect the morphologic changes of the apoptotic cells. The expression of LMP1/miR-155/p65/pp65/PU.1 was evaluated by quantitative real-time PCR (qRT-PCR) and western blot, while that of NF-κB was analyzed by EMSA. At last, immunohistochemical staining was applied to assess the expression of target proteins and relevant molecules.
In vitro, wogonin induced the apoptosis of Raji cells by downregulating the expression of NF-κB through LMP1/miR-155/NF-κB/PU.1 pathway, which was in a dose and time-dependent manner. In vivo, wogonin could suppress tumor growth, associated with the downregulation of ki67, p65 and upregulation of PU.1.
Wogonin could suppress tumor growth and induce cell apoptosis by inhibiting the expression of NF-κB. Taken these findings, we concluded that wogonin could be a potential targeted therapeutic agent for EBV-positive lymphoma with the expression of LMP1 through the pathway of LMP1/NF-κB/miR-155/PU.1.
汉黄芩素因其强大的抗肿瘤和抗炎作用以及高安全性，是临床应用中一个令人鼓舞的选择。然而，汉黄芩素用于淋巴瘤靶向治疗的研究尚不充分。在本研究中，我们聚焦于其抗癌作用以及EBV阳性淋巴瘤靶向治疗的潜在机制。这将有助于其在不久的将来引入临床应用。
采用CCK8法研究细胞增殖。运用流式细胞术分析细胞凋亡和细胞周期阻滞。此外，我们还使用免疫荧光染色检测凋亡细胞的形态变化。通过定量实时PCR（qRT-PCR）和蛋白质免疫印迹法评估LMP1/miR-155/p65/pp65/PU.1的表达，同时通过电泳迁移率变动分析（EMSA）分析NF-κB的表达。最后，应用免疫组织化学染色评估靶蛋白和相关分子的表达。
在体外，汉黄芩素通过LMP1/miR-155/NF-κB/PU.1通路下调NF-κB的表达，以剂量和时间依赖性方式诱导Raji细胞凋亡。在体内，汉黄芩素可抑制肿瘤生长，这与ki67、p65的下调以及PU.1的上调有关。
汉黄芩素可通过抑制NF-κB的表达抑制肿瘤生长并诱导细胞凋亡。基于这些发现，我们得出结论，汉黄芩素可能是一种通过LMP1/NF-κB/miR-155/PU.1通路对表达LMP1的EBV阳性淋巴瘤具有潜在靶向治疗作用的药物。
