Institute of Neurological Disease, West China Hospital, Sichuan University, Chengdu 610041, Sichuan, China.
Department of Anesthesiology, Affiliated Traditional Chinese Medicine Hospital, Southwest Medical University, Luzhou 646000, Sichuan, China.
Biomed Res Int. 2020 Oct 10;2020:9786428. doi: 10.1155/2020/9786428. eCollection 2020.
Colorectal cancer (CRC) is an underlying deadly malignancy with poor prognosis, lacking effective therapies currently available to improve the prognosis. CHNO (AUCAN), a kind of dibenzofuran extracted from a special plant in Yunnan Province (China), is identified as a natural anticancer agent exerting strong inhibitory activities on various cancers. Our study was committed to investigating the potency of AUCAN against colorectal cancers and further exploring the potential mechanisms via proteomic analysis.
Cell Counting Kit-8 assay and immunofluorescence staining were used to investigate the effect of AUCAN on the viability and proliferation of HCT-116 cells and RKO cells. The apoptosis of HCT-116 and RKO cells after AUCAN administration was determined by the flow cytometry test. The effects of AUCAN on invasion and migration of tumor cells were investigated by the colony formation assay, wound healing test, and Transwell invasion test. Meanwhile, the energy metabolism and growth of tumor tissues after AUCAN administration with 10 mg/kg and 20 mg/kg were examined by PET-CT . The side effects of AUCAN treatment were also evaluated through blood routine and liver function examination. RKO cell proliferation and apoptosis were further determined by hematoxylin and eosin staining, TUNEL staining, and immunohistochemistry. Furthermore, the differentially expressed proteins (DEPs) involved in AUCAN treatment were determined by proteomic analysis followed by functional clustering analysis.
The results showed that AUCAN suppressed the migratory abilities and enhanced apoptosis of HCT-116 and RKO cell lines. Meanwhile, AUCAN treatment dramatically depressed the growth and volume of colorectal tumors in nude mice and suppressed the survival of RKO cells in tumor tissues without any side effects on the blood routine and liver function. In addition, twenty-four upregulated and forty-two downregulated proteins were identified. Additionally, functional clustering analysis concealed enriched biological processes, cellular components, molecular functions, and related pathways of these proteins involved in cellular metabolic. Finally, the protein-protein interaction analysis revealed the regulatory connection among these DEPs.
Taken together, AUCAN exerted its significant antitumor effect without side effects in the blood routine and liver function and the underlying mechanisms were preliminarily investigated by proteomic analysis.
结直肠癌（CRC）是一种预后不良的潜在致命恶性肿瘤，目前缺乏有效的治疗方法来改善预后。CHNO（AUCAN）是从中国云南省特殊植物中提取的一种二苯并呋喃，被鉴定为一种天然抗癌剂，对各种癌症具有很强的抑制活性。我们的研究致力于研究 AUCAN 对结直肠癌的效力，并通过蛋白质组学分析进一步探讨潜在的机制。
使用细胞计数试剂盒-8 测定法和免疫荧光染色法研究 AUCAN 对 HCT-116 细胞和 RKO 细胞活力和增殖的影响。通过流式细胞术试验测定 AUCAN 给药后 HCT-116 和 RKO 细胞的凋亡情况。通过集落形成试验、划痕愈合试验和 Transwell 侵袭试验研究 AUCAN 对肿瘤细胞侵袭和迁移的影响。同时，通过 PET-CT 检查 AUCAN 给药后 10mg/kg 和 20mg/kg 对肿瘤组织的能量代谢和生长的影响。还通过血常规和肝功能检查评估 AUCAN 治疗的副作用。通过苏木精和伊红染色、TUNEL 染色和免疫组化进一步确定 RKO 细胞增殖和凋亡。此外，通过蛋白质组学分析和功能聚类分析确定参与 AUCAN 治疗的差异表达蛋白（DEPs）。
结果表明，AUCAN 抑制了 HCT-116 和 RKO 细胞系的迁移能力并增强了其凋亡能力。同时，AUCAN 治疗显著抑制了裸鼠结直肠肿瘤的生长和体积，并抑制了肿瘤组织中 RKO 细胞的存活，而对血常规和肝功能无任何副作用。此外，鉴定出 24 个上调和 42 个下调蛋白。此外，功能聚类分析揭示了这些蛋白涉及细胞代谢的丰富生物学过程、细胞成分、分子功能和相关途径。最后，蛋白质-蛋白质相互作用分析揭示了这些 DEPs 之间的调节关系。
总之，AUCAN 在血常规和肝功能方面无副作用，具有显著的抗肿瘤作用，并通过蛋白质组学分析初步探讨了其潜在的作用机制。
