The Third Affiliated Hospital of Zunyi Medical University (The First People's Hospital of Zunyi), Central Laboratory, Guizhou, 563002, PR China.
College of Medicine, Yanan University, Yanan, 716000, PR China.
Biomed Pharmacother. 2020 Aug;128:110322. doi: 10.1016/j.biopha.2020.110322. Epub 2020 Jun 4.
Streptomyces Sp FJS31-2 is a strain isolated from special habitat soils in the early stage of our laboratory for producing a new type of halogenated type II polyketide antibiotic with good anti-MRSA activity. In this experiment, a variety of chromatographic and spectroscopic methods was used to isolate and identify a milbemycin compound VM48130 from the ethyl acetate extract of the fermentation products. To investigate its bioactivity, Cell Counting Kit-8 (CCK-8) assay was used to test the cytotoxic activity of the compound against a variety of cancer cells (human liver cancer cell line MHCC97H and SK-Hep1, human nasopharyngeal carcinoma cell line CNE1, mouse melanoma cell line B16, human colon cancer cell line LOVO, human lung adenocarcinoma cell line A549) and normal cells (human bronchial epithelial cell line 16HBE, human normal liver cell line L02, human nasopharyngeal epithelial cell line NP69). The results showed that the compound had significant cytotoxic activity against the above cancer cells, and the IC values were 21.96 ± 1.45, 22.18 ± 0.55, 19.42 ± 0.71, 18.61 ± 1.68, 18.62 ± 0.67, 18.52 ± 0.64 μM, respectively. Furthermore, the CCK-8 method was used to evaluate the compound's reversal of cisplatin resistance in multidrug resistant cisplatin-resistant human lung adenocarcinoma (A549/DDP) cells. The results indicated that when the compound concentration was 0.5 μM, the reversal fold (RF) reached 6.25 and showed a dose-dependent effect. At 5 μM, the RF reached 8.35, which was approximately equivalent to the reversal effect of the positive drug verapamil at the same concentration. The expression of MDR1, MRP1, LRP, MAST1 resistance genes and the corresponding proteins were analyzed by quantitative RT-PCR and Western blot assay, and found that the compound could significantly down-regulate the expression of these genes and proteins. These results indicated that VM48130 had the potential of being a lead compound for the treatment or adjuvant treatment of cancer.
链霉菌 Sp FJS31-2 是本实验室在早期从特殊生境土壤中分离得到的一株菌，用于生产具有良好抗 MRSA 活性的新型卤代 II 型聚酮类抗生素。本实验采用多种色谱和光谱方法，从发酵产物的乙酸乙酯提取物中分离鉴定出米尔贝霉素化合物 VM48130。为了研究其生物活性，采用细胞计数试剂盒（CCK-8）法检测化合物对多种癌细胞（人肝癌细胞系 MHCC97H 和 SK-Hep1、人鼻咽癌细胞系 CNE1、小鼠黑色素瘤细胞系 B16、人结肠癌细胞系 LOVO、人肺腺癌细胞系 A549）和正常细胞（人支气管上皮细胞系 16HBE、人正常肝细胞系 L02、人鼻咽上皮细胞系 NP69）的细胞毒性。结果表明，该化合物对上述癌细胞具有显著的细胞毒性，IC 值分别为 21.96±1.45、22.18±0.55、19.42±0.71、18.61±1.68、18.62±0.67、18.52±0.64 μM。此外，采用 CCK-8 法评价该化合物对多药耐药顺铂耐药人肺腺癌细胞（A549/DDP）中顺铂耐药的逆转作用。结果表明，当化合物浓度为 0.5 μM 时，逆转倍数（RF）达到 6.25，呈现浓度依赖性。当浓度为 5 μM 时，RF 达到 8.35，与相同浓度的阳性药物维拉帕米的逆转效果相当。采用定量 RT-PCR 和 Western blot 检测分析 MDR1、MRP1、LRP、MAST1 耐药基因及其相应蛋白的表达，发现该化合物可显著下调这些基因和蛋白的表达。这些结果表明，VM48130 具有成为治疗或辅助治疗癌症的潜在先导化合物的潜力。
