Community Health Service Center, Shenzhen Bao'an Traditional Chinese Medicine Hospital Group, 25 yu'an 2nd Road, Baoan District, Shenzhen, Guangdong, China.
Department of Traditional Chinese Medicine, The Second Affiliated Hospital of Shenzhen University (People's Hospital of Shenzhen Baoan District), 118 Longjing 2nd Road, Baoan District, Shenzhen, 518100, Guangdong, China.
Mol Cell Biochem. 2024 Aug;479(8):2131-2141. doi: 10.1007/s11010-023-04836-7. Epub 2023 Aug 28.
The purpose of this study was to demonstrate the regulatory effect of berberine (BBR) on the intestinal microbiota and related epigenetics during the inhibition of colon cancer cell growth in vitro and in vivo. We used a nude mouse xenograft model with HT29 colon cancer cells to establish and divide into a model group and BBR group. The mice were treated for four weeks, and HT29 cells in the BBR group were cultured for 48 h. Cetuximab and the DNA transmethylase (DNMT) inhibitor 5-AZA-dC were added to HT29 cells. Tumour volume and weight were measured by hematoxylin-eosin (HE) staining for histopathological observation. Mouse faeces were collected, and the gut microbiota was analysed with 16S rDNA amplicons. The levels of cytokines in the supernatant of HT29 cells were measured by ELISA. A CCK-8 kit was used to examine the proliferation of HT29 cells, and RT‒PCR was used to measure the levels of c-Myc, DNMT1, DNMT3A, and DNMT3B. We found that BBR reduced the growth of colon cancer cells to a certain extent in vitro and in vivo, although the difference was not statistically significant compared with that in the model group. BBR significantly mediated the abundance, composition and metabolic functions of the intestinal microbial flora in mice with colon cancer. The effect of BBR on inflammatory cytokines, including IL-6, FGF, and PDGF, was not obvious, but BBR significantly downregulated IL-10 levels (P < 0.05) and reduced c-Myc, DNMT1, and DNMT3B levels (P < 0.05). Inhibiting DNMTs with 5-AZA-dC significantly suppressed the proliferation of HT29 cells, which was consistent with the effect of BBR. The inhibitory effect of berberine on colon cancer is related not only to the intestinal microbiota and its metabolic functions but also to the regulation of DNMTs.
本研究旨在展示小檗碱（BBR）在体外和体内抑制结肠癌细胞生长过程中对肠道微生物群及其相关表观遗传学的调节作用。我们使用裸鼠 HT29 结肠癌异种移植模型进行建立并分为模型组和 BBR 组。小鼠治疗四周，BBR 组 HT29 细胞培养 48 小时。向 HT29 细胞中加入西妥昔单抗和 DNA 甲基转移酶（DNMT）抑制剂 5-AZA-dC。通过苏木精-伊红（HE）染色测量肿瘤体积和重量，进行组织病理学观察。收集小鼠粪便，用 16S rDNA 扩增子分析肠道微生物群。通过 ELISA 测量 HT29 细胞上清液中细胞因子的水平。使用 CCK-8 试剂盒检测 HT29 细胞的增殖，使用 RT-PCR 测量 c-Myc、DNMT1、DNMT3A 和 DNMT3B 的水平。我们发现 BBR 在体外和体内在一定程度上减少了结肠癌的生长，尽管与模型组相比差异无统计学意义。BBR 显著调节了结肠癌小鼠肠道微生物群落的丰度、组成和代谢功能。BBR 对包括 IL-6、FGF 和 PDGF 在内的炎症细胞因子的作用不明显，但 BBR 显著降低了 IL-10 水平（P<0.05）并降低了 c-Myc、DNMT1 和 DNMT3B 水平（P<0.05）。用 5-AZA-dC 抑制 DNMTs 显著抑制了 HT29 细胞的增殖，这与 BBR 的作用一致。小檗碱对结肠癌的抑制作用不仅与肠道微生物群及其代谢功能有关，还与 DNMTs 的调节有关。
