Xu Dong-bo, Wang Yun-ling, Yue Yuan, Wu Shuang-chan, Ding Hong
Department of Pharmacology, Wuhan University of Pharmaceutical Sciences, Wuhan 430072, China.
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Zhonghua Zhong Liu Za Zhi. 2013 Nov;35(11):814-8.
To investigate the inhibitory effects of a novel histone deacetylases inhibitor FK228 on human colon cancer HCT-116 cells in vitro and in vivo, and evaluate its toxicity and side effects.
The in vitro growth inhibitions of HCT-116 cells by different concentrations of FK228 and 5-Fu for 24, 48 and 72 h were assessed by CCK-8 assay. BALB/c nude mouse models of tumor xenografts were prepared by subcutaneous implantation of tumor tissue, and 4 mg/kg FK228 and 50 mg/kg 5-Fu were i.p. injected, respectively. The inhibitory effects on tumor growth, hematology, and liver and kidney function were evaluated.
CCK-8 assay indicated that FK228 had an obvious growth inhibitory effect on HCT-116 cells in a dose- and time-dependent manner. The IC50 of FK228 in HCT-116 cells was 12.05 ng/ml for 48 h, while the IC50 of 5-Fu was 18.92 µg/ml. At 20 days after FK228 and 5-Fu treatment, the tumor volume of the FK228 group was (139.71 ± 44.54)mm(3), significantly lower than that of the 5-Fu group [(282.28 ± 58.81)mm(3)] and that of the model group [(520.65 ± 39.73)mm(3), P < 0.01 for both]. The average tumor weight was (0.07 ± 0.02)g in the FK228 group, significantly lower than that of the 5-Fu group [(0.20 ± 0.08)g, P < 0.01]. The tumor growth inhibition rate of the FK228 group was 73.2%, significantly higher than that of the 5-Fu group (45.8%, P < 0.01). The ALT levels of the FK228 and 5-Fu groups were significantly higher than that of the model group (P < 0.01). The BUN of 5-Fu group was significantly higher than that of the model group (P < 0.01), but the BUN of FK228 group was not significantly different from that of the blank and control groups (P > 0.05 for both). Routine blood test showed that WBC, RBC, Hb and PLT of the 5-Fu group were significantly lower than those of the model group (P < 0.05 for all), but only WBC of the FK228 group was significantly lower than that of the model group (P < 0.05). The pathological examination using HE staining revealed that in the FK228 group, there were fibrosis and inflammatory cell infiltration in the liver tissue, and mild edema of the renal tubules in the kidney. However, in the 5-Fu group there were extensive hepatocyte edema and necrosis in the liver, and evident deformation and necrosis of glomeruli and tubules, and tubular wall thinning in the kidney.
The results of this study indicate that FK228 can more effectively than 5-Fu inhibit the growth of HCT-116 cells in vitro and vivo, and without obvious toxic effect on the kidney and hematology. Its clinical value in colon cancer treatment deserves further investigation.
研究新型组蛋白去乙酰化酶抑制剂FK228对人结肠癌HCT - 116细胞的体内外抑制作用，并评估其毒性和副作用。
采用CCK - 8法检测不同浓度的FK228和5 - 氟尿嘧啶（5 - Fu）作用于HCT - 116细胞24、48和72 h后的体外生长抑制情况。通过皮下接种肿瘤组织制备BALB/c裸鼠肿瘤异种移植模型，分别腹腔注射4 mg/kg FK228和50 mg/kg 5 - Fu，评估其对肿瘤生长、血液学指标以及肝肾功能的抑制作用。
CCK - 8法检测结果表明，FK228对HCT - 116细胞具有明显的生长抑制作用，呈剂量和时间依赖性。FK228作用于HCT - 116细胞48 h的半数抑制浓度（IC50）为12.05 ng/ml，而5 - Fu的IC50为18.92 μg/ml。FK228和5 - Fu处理20天后，FK228组肿瘤体积为（139.71±44.54）mm³，显著低于5 - Fu组[（282.28±58.81）mm³]和模型组[（520.65±39.73）mm³，两者P均<0.01]。FK228组平均肿瘤重量为（0.07±0.02）g，显著低于5 - Fu组[（0.20±0.08）g，P<0.01]。FK228组肿瘤生长抑制率为73.2%，显著高于5 - Fu组（45.8%，P<0.01）。FK228组和5 - Fu组谷丙转氨酶（ALT）水平均显著高于模型组（P<0.01）。5 - Fu组血尿素氮（BUN）显著高于模型组（P<0.01），而FK228组BUN与空白组和对照组相比差异无统计学意义（两者P>0.05）。血常规检测显示，5 - Fu组白细胞（WBC）、红细胞（RBC）、血红蛋白（Hb）和血小板（PLT）均显著低于模型组（均P<0.05），而FK228组仅WBC显著低于模型组（P<0.05）。苏木精 - 伊红（HE）染色病理检查显示，FK228组肝组织有纤维化和炎性细胞浸润，肾组织肾小管轻度水肿；而5 - Fu组肝组织有广泛的肝细胞水肿和坏死，肾组织肾小球和肾小管明显变形、坏死，肾小管壁变薄。
本研究结果表明，FK228在体外和体内均能比5 - Fu更有效地抑制HCT - 116细胞生长，且对肾脏和血液学无明显毒性作用。其在结肠癌治疗中的临床价值值得进一步研究。
