桑树提取物桑根酮C影响神经胶质母细胞瘤细胞生长增殖的初步研究

Effect of Morus alba extract sanggenon C on growth and proliferation of glioblastoma cells

胶质母细胞瘤是最常见的原发性颅脑恶性肿瘤,化学药物治疗仍然是其治疗的重要手段。桑根酮C (sanggenon C, San C)是一类从桑属植物中提取的天然黄酮类化合物,是一种潜在的抗肿瘤中药单体。该研究采用methyl thiazolyl tetrazolium(MTT)实验、5-bromodeoxyuridinc(BrdU)标记实验检测San C对胶质母细胞瘤细胞生长增殖能力的影响;采用流式细胞术检测San C对肿瘤细胞周期的影响;采用软琼脂实验检测San C对肿瘤细胞形成克隆和自我更新能力的影响;采用Western blot和生物信息学分析初步探究San C的抗肿瘤活性的作用机制。在San C的作用下,MTT实验表明San C以剂量和时间依赖性的方式显著地抑制肿瘤细胞的生长增殖;BrdU标记实验显示San C可显著减弱肿瘤细胞核内DNA复制的活性;流式细胞术则证实桑根酮C将肿瘤细胞的细胞周期阻滞于G_(0)/G_(1)期;软琼脂克隆形成实验揭示San C显著减弱肿瘤细胞的克隆形成和自我更新的能力;基因集富集分析(gene set enrichment analysis, GSEA)提示San C是通过影响myelocytomatosis viral oncogene(MYC)信号通路进而抑制肿瘤细胞分裂周期的;Western blot实验揭示San C是通过lysine demethylase 4B(KDM4B)调控MYC信号通路抑制周期蛋白的表达,最终抑制神经胶质母细胞瘤细胞的生长增殖和自我更新。总之,San C可通过靶标KDM4B-MYC轴抑制神经胶质母细胞瘤细胞生长增殖和自我更新的能力,表现出潜在的抗肿瘤活性。

Glioblastoma is the most common primary cranial malignancy, and chemotherapy remains an important tool for its treatment. Sanggenon C(San C), a class of natural flavonoids extracted from Morus plants, is a potential antitumor herbal monomer. In this study, the effect of San C on the growth and proliferation of glioblastoma cells was examined by methyl thiazolyl tetrazolium(MTT) assay and 5-bromodeoxyuridinc(BrdU) labeling assay. The effect of San C on the tumor cell cycle was examined by flow cytometry, and the effect of San C on clone formation and self-renewal ability of tumor cells was examined by soft agar assay. Western blot and bioinformatics analysis were used to investigate the mechanism of the antitumor activity of San C. In the presence of San C, the MTT assay showed that San C significantly inhibited the growth and proliferation of tumor cells in a dose and time-dependent manner. BrdU labeling assay showed that San C significantly attenuated the DNA replication activity in the nucleus of tumor cells. Flow cytometry confirmed that San C blocked the cell cycle of tumor cells in G_(0)/G_(1) phase. The soft agar clone formation assay revealed that San C significantly attenuated the clone formation and self-renewal ability of tumor cells. The gene set enrichment analysis(GSEA) implied that San C inhibited the tumor cell division cycle by affecting the myelocytomatosis viral oncogene(MYC) signaling pathway. Western blot assay revealed that San C inhibited the expression of cyclin through the regulation of the MYC signaling pathway by lysine demethylase 4B(KDM4B), which ultimately inhibited the growth and proliferation of glioblastoma cells and self-renewal. In conclusion, San C exhibits the potential antitumor activity by targeting the KDM4B-MYC axis to inhibit glioblastoma cell growth, proliferation, and self-renewal.