Zhong Yu, Zheng Xue-Bao, Cai Kang-Rong, Huang Qiong-Lin, Ye Hua, Huang Zhe, Wang Yong-Cun, Zhu Yu-Zhen
Analysis Center of Guangdong Medical University, Zhanjiang 524023, China.
Mathematical Engineering Academy of Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou 510006, China.
Zhongguo Zhong Yao Za Zhi. 2018 Jun;43(11):2358-2364. doi: 10.19540/j.cnki.cjcmm.20180130.001.
To investigate the inhibitory effect of isobutyrylshikonin on the growth of human colon carcinoma cells and its effect on the PI3K/Akt/m-TOR pathway. MTT assay was used to detect the inhibitory effect of different concentrations (0, 6.25, 12.5, 25, 50, 100 mg·L⁻¹) of isobutyrylshikonin on the proliferation of human colon carcinoma cell HT29 at 24, 48 h. CCK-8 method was used to detect the inhibitory effect of isobutyrylshikonin on HT29, HCT116, DLD-1 and Caco-2 at 48 h. AnnexinV/propidium iodide staining was applied in detecting the apoptoticrate of HT29 cells treated with different concentrations of isobutyrylshikonin at 24 h and 48 h. Cycletest plus DNA was employed to analyze HT29 apoptosis and cell cycle after 48 h treatment with isobutyrylshikonin at different concentrations. Western blot and RT-PCR assay were used to examine the protein and mRNA expressions of PI3K, p-PI3K, Akt, p-Akt and m-TOR. The results showed that isobutyrylshikonin inhibited the proliferation of different human colon carcinoma cells, and the inhibition rate was in a dose-dependent manner. Isobutyrylshikonin induced apoptosis mainly in the early stage and blocked cells in the G₀/G₁ or G₂/M phase. Isobutyrylshikonin reduced the protein expressions of PI3K, p-PI3K, Akt, p-Akt, m-TOR and the mRNA expressions of PI3K, Akt, m-TOR in a dose-dependent manner. Isobutyrylshikonin can significantly inhibit the proliferation, induce the early apoptosis and change the cycle distribution in colon carcinoma cells.This biological effect may be correlated with the inhibition of PI3K/AKT/m-TOR pathway.
研究异丁酰紫草素对人结肠癌细胞生长的抑制作用及其对PI3K/Akt/m-TOR信号通路的影响。采用MTT法检测不同浓度（0、6.25、12.5、25、50、100 mg·L⁻¹）的异丁酰紫草素在24、48 h对人结肠癌细胞HT29增殖的抑制作用。采用CCK-8法检测异丁酰紫草素在48 h对HT29、HCT116、DLD-1和Caco-2细胞的抑制作用。应用AnnexinV/碘化丙啶染色检测不同浓度异丁酰紫草素处理24 h和48 h后HT29细胞的凋亡率。采用Cycletest plus DNA分析不同浓度异丁酰紫草素处理48 h后HT29细胞的凋亡及细胞周期情况。采用蛋白质免疫印迹法（Western blot）和逆转录-聚合酶链反应（RT-PCR）检测PI3K、p-PI3K、Akt、p-Akt和m-TOR的蛋白及mRNA表达。结果显示，异丁酰紫草素可抑制不同人结肠癌细胞的增殖，且抑制率呈剂量依赖性。异丁酰紫草素主要诱导早期凋亡，并将细胞阻滞于G₀/G₁期或G₂/M期。异丁酰紫草素可呈剂量依赖性降低PI3K、p-PI3K、Akt、p-Akt、m-TOR的蛋白表达以及PI3K、Akt、m-TOR的mRNA表达。异丁酰紫草素可显著抑制结肠癌细胞的增殖，诱导早期凋亡并改变细胞周期分布。这种生物学效应可能与抑制PI3K/AKT/m-TOR信号通路有关。
