本研究旨在探讨一种多草药制剂Habb-e-Ustukhuddus（HU）的抗癌和化学预防性能。HU被广泛用于其抗炎作用，然而其抗癌和化学预防作用尚未知晓，因此本研究对其进行了详细调查。本实验将癌细胞分别处理50-400 µg/ml的HU，对细胞进行MTT、尝试蓝和克隆形成实验。流式细胞仪用于细胞周期进程、细胞凋亡和线粒体膜电位分析，包括碘化丙啶（PI）染色、annexin V-FITC实验和JC-1染色。此外，还进行了免疫印记、细胞迁移和侵袭实验，以及HU的化学特征分析，包括气相色谱-质谱和高效液相色谱。我们使用C57BL/6小鼠评估了HU的体内毒性。在评估抗癌活性时，HU的甲醇提取物（50-400 µg/ml）强烈抑制了肺癌和乳腺癌细胞的生长和存活（P<0.05-0.001），并导致细胞人口在G1期后期增加。HU引起了细胞凋亡（P<0.05-0.001），与线粒体膜电位（Δψ）的去极化（P<0.001）和Bax到Bcl-2蛋白比例的增加有关。此外，HU抑制了癌细胞的侵袭和迁移，伴随着上皮标志物E-钙黏蛋白的增加和间充质标志物vimentin的减少。HU通过GC-MS和HPLC分析的特征化显示了大量生物活性化合物，包括黄酮类物质和生物碱。在化学预防研究中，将HU的甲醇提取物口服给小鼠（剂量为50和100 mg/kg体重），未引起任何毒性反应，并显著增加了肝脏药物代谢相I和相II酶的特异性活性，表明具有解毒外源性化合物的潜力。综上所述，这些结果证明了HU的抗癌潜力，在小鼠中没有明显的毒性反应，因此可进一步探索其在癌症控制中的临床应用前景。

A polyherbal medicine, Habb-e-Ustukhuddus (HU), is used for its anti-inflammatory properties. However, the anticancer and chemopreventive properties of HU were not known, and Therefore, investigated in  the  present study.Cancer cells were treated with 50-400 µg/ml HU and MTT, trypan blue, and clonogenic assays were performed. Propidium iodide (PI) staining, annexin V-FITC assay, and JC-1 staining were done for cell cycle progression, apoptosis, and mitochondrial membrane potential, respectively, using flow cytometry. Immunoblotting, cell migration and invasion assays were performed. Chemical characterization of HU was done through GC-MS and HPLC analyses. C57BL/6 mice were used to assess the in vivo toxicity of HU.While evaluating the anticancer activity, the methanolic extract of HU (50-400 µg/ml) strongly inhibited the growth and survival (P<0.05-0.001) of lung and breast cancer cells and increased the cell population in the sub-G1 phase of the cell cycle. HU caused apoptotic death of cancer cells (P<0.05-0.001), which was associated with the depolarization of mitochondrial membrane potential (Δψ) (P<0.001) and an increase in Bax to Bcl-2 protein ratio. Further, HU inhibited the invasion and migration of cancer cells, which was accompanied by an increase in the epithelial marker, E-cadherin, and a decrease in the mesenchymal marker, vimentin. The HU characterization by GC-MS and HPLC analyses showed the abundance of bioactive compounds including flavonoids and alkaloids. In the chemopreventive study, the oral administration of methanolic extract of the formulation HU (50 and 100 mg/kg body weight) to mice did not cause any toxicity and significantly increased the specific activities of hepatic drug metabolizing phase I and phase II enzymes, which suggested for its detoxification potential of xenobiotic compounds.Together, these results demonstrated the anticancer potential HU, without any apparent toxicity in mice, and thus HU could be further explored for its clinical utility in cancer control.