School of Pharmacy, Ningxia Medical University, Yinchuan 750004, China.
School of Basic Science, Ningxia Medical University, Yinchuan 750004, China.
Curr Cancer Drug Targets. 2022;22(11):889-903. doi: 10.2174/1568009622666220627160834.
Colon cancer is a gastrointestinal malignancy with high incidence and poor prognosis.
Saikosaponin B4 (SSB4) is a monomeric component of the Traditional Chinese medicine (TCM), Bupleurum. The current study investigates the therapeutic effect and mechanisms of SSB4 in colon cancer.
The proliferation of two colon cancer cell lines, SW480 and SW620, were assessed using CCK8 and expression of regulatory molecules, including Bax, Caspase3, Caspase9, Cleaved Caspase3, Cleaved Caspase9 and Bcl2 by flow cytometry and Western blotting.
Survival rates, assessed by CCK8, of SW480 and SW620 cells decreased significantly when the SSB4 concentration was in the range 12.5-50 μg/ml. Flow cytometry measurements indicated apoptosis rates of 55.07% ± 1.63% for SW480 cells and 33.07% ± 1.28% for SW620 cells treated with 25 μg/ml SSB4. Western blotting revealed upregulation of the proapoptotic proteins, Bax, Caspase3, Caspase9, Cleaved Caspase3 and Cleaved Caspase9, and downregulation of the anti-apoptotic protein, Bcl2, in the presence of SSB4. Network pharmacology and molecular docking predicted that the PI3K/Akt/mTOR pathway might be the main regulatory target for the antitumor effect of SSB4. Further Western blotting experiments showed that SSB4 downregulated (p < 0.01) expression of PI3K, Akt, mTOR and the phosphorylated proteins, P-PI3K, P-Akt and P-MTOR. Expression of PI3K, Akt and mTOR mRNA was found to be downregulated by SSB4 (P < 0.01) as the result of RT-PCR measurements.
SSB4 is a potent anti-colon cancer agent. Its effects are likely to be mediated by suppression of the PI3K/AKT/mTOR pathway.
结肠癌是一种发病率高、预后差的胃肠道恶性肿瘤。
柴胡皂苷 B4（SSB4）是一种来源于中药柴胡的单体成分。本研究旨在探讨 SSB4 对结肠癌的治疗作用及机制。
通过 CCK8 检测两种结肠癌细胞系 SW480 和 SW620 的增殖情况，流式细胞术检测凋亡相关调控分子 Bax、Caspase3、Caspase9、Cleaved Caspase3、Cleaved Caspase9 和 Bcl2 的表达，Western blot 检测凋亡相关调控分子的表达。
CCK8 结果显示，SSB4 浓度在 12.5-50 μg/ml 范围内时，SW480 和 SW620 细胞的存活率显著降低。流式细胞术检测结果显示，浓度为 25 μg/ml 的 SSB4 处理的 SW480 细胞凋亡率为 55.07%±1.63%，SW620 细胞凋亡率为 33.07%±1.28%。Western blot 结果显示，SSB4 处理后促凋亡蛋白 Bax、Caspase3、Caspase9、Cleaved Caspase3 和 Cleaved Caspase9 的表达上调，抗凋亡蛋白 Bcl2 的表达下调。网络药理学和分子对接预测，PI3K/Akt/mTOR 通路可能是 SSB4 抗肿瘤作用的主要调控靶点。进一步的 Western blot 实验结果显示，SSB4 下调了 PI3K、Akt、mTOR 及其磷酸化蛋白 P-PI3K、P-Akt 和 P-MTOR 的表达（p<0.01）。RT-PCR 结果显示，SSB4 下调了 PI3K、Akt 和 mTOR mRNA 的表达（p<0.01）。
SSB4 是一种有效的抗结肠癌药物，其作用可能是通过抑制 PI3K/AKT/mTOR 通路来实现的。
