Hussain Yusuf, Singh Jyoti, Meena Abha, Sinha Rohit Anthony, Luqman Suaib
Bioprospection and Product Development Division, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow, India.
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, India.
Environ Toxicol. 2024 Feb;39(2):840-856. doi: 10.1002/tox.23988. Epub 2023 Oct 18.
Hepatocellular carcinoma (HCC) is a common solid cancer and the leading cause of cancer deaths worldwide. Sorafenib is the first drug used to treat HCC but its effectiveness needs to be improved, and it is important to find ways to treat cancer that combine sorafenib with other drugs. Synergistic therapies lower effective drug doses and side effects while enhancing the anticancer effect.
In the present study, the therapeutic potential of sorafenib in combination with escin and its underlying mechanism in targeting liver cancer has been established.
STUDY DESIGN/METHODS: The IC of sorafenib and escin against HepG2, PLC/PRF5 and Huh7 cell lines were determined using MTT assay. The combination index, dose reduction index, isobologram and concentrations producing synergy were evaluated using the Chou-Talaly algorithm. The sub-effective concentration of sorafenib and escin was selected to analyze cytotoxic synergistic potential. Cellular ROS, mitochondrial membrane potential, annexin V and cell cycle were evaluated using a flow-cytometer, and autophagy biomarkers were determined using western blotting. Moreover, autophagy was knocked down using ATG5 siRNA to confirm its role. A DEN-induced liver cancer rat model was developed to check the synergy of sorafenib and escin.
Different concentrations of escin reduced the IC of sorafenib in HepG2, PLC/PRF5 and Huh7 cell lines. Chou-Talaly algorithm determined cytotoxic synergistic concentrations of sorafenib and escin in these cell lines. Mechanistically, this combination over-expressed p62 and LC-II, reflecting autophagy block and induced late apoptosis, further reconfirmed by ATG5 knockdown. Sorafenib and escin combination reduced HCC serum biomarker α-feto protein (α-FP) by 1.5 folds. This combination restricted liver weight, tumor number and size, also, conserved morphological features of liver cells. The combination selectively targeted the G /G phase of cancer cells.
Escin and sorafenib combination potentially up-regulates p62 to block autophagy to induce late apoptosis in liver cancer cells.
肝细胞癌（HCC）是一种常见的实体癌，也是全球癌症死亡的主要原因。索拉非尼是第一种用于治疗HCC的药物，但其疗效有待提高，因此找到将索拉非尼与其他药物联合治疗癌症的方法非常重要。协同疗法在增强抗癌效果的同时降低了有效药物剂量和副作用。
在本研究中，已确定索拉非尼与七叶皂苷联合使用的治疗潜力及其靶向肝癌的潜在机制。
研究设计/方法：使用MTT法测定索拉非尼和七叶皂苷对HepG2、PLC/PRF5和Huh7细胞系的半数抑制浓度（IC）。使用Chou-Talaly算法评估联合指数、剂量降低指数、等效线图和产生协同作用的浓度。选择索拉非尼和七叶皂苷的亚有效浓度来分析细胞毒性协同潜力。使用流式细胞仪评估细胞活性氧（ROS）、线粒体膜电位、膜联蛋白V和细胞周期，并使用蛋白质免疫印迹法测定自噬生物标志物。此外，使用ATG5小干扰RNA（siRNA）敲低自噬以确认其作用。建立二乙基亚硝胺（DEN）诱导的肝癌大鼠模型以检查索拉非尼和七叶皂苷的协同作用。
不同浓度的七叶皂苷降低了索拉非尼在HepG2、PLC/PRF5和Huh7细胞系中的IC。Chou-Talaly算法确定了索拉非尼和七叶皂苷在这些细胞系中的细胞毒性协同浓度。从机制上讲，这种联合上调了p62和LC-II，反映出自噬阻滞并诱导晚期凋亡，ATG5敲低进一步证实了这一点。索拉非尼和七叶皂苷联合使用使肝癌血清生物标志物甲胎蛋白（α-FP）降低了1.5倍。这种联合限制了肝脏重量、肿瘤数量和大小，同时保留了肝细胞的形态特征。该联合选择性地靶向癌细胞的G1/G0期。
七叶皂苷和索拉非尼联合使用可能上调p62以阻断自噬，从而诱导肝癌细胞晚期凋亡。
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