Screening Effective Constituent Of Lingjia Anticancer Medicine Targeting To NF-κB

Lingjia Anticancer Medicine (LAM) is a Traditional Chinese Medicine formula which showed positive efficacy on liver cancer and lung cancer in clinical treatment, but the mechanism of its antitumor effects is still unclear. Ingredients of Traditional Chinese Medicine are complicated and effective constituent and efficacy of Traditional Chinese Medicine closely associated with the process of processing and decoction. The NF-κB signaling pathway plays a major role in the development, maintenance, and progression of most diseases. Mounting evidence indicates that NF-κB acts as a important link between inflammation and cancer progression, making NF-κB to a potential drug target in treatment of hematological malignancies and solid tumors. According to recent studies, high-throughput screening targeting to NF-κB has become an effective means for discovery of anticancer drug.The aim of our study is to explore the effective constituent and molecular mechanism of Lingjia Anticancer Medicine by evaluating its effects on NF-κB activity. The LAM were decocted in traditional method, and then LAM were extracted by45%ethanol,80%ethanol and H2O. Extracts of LAM were eparately collected and were separately named as LAM-1, LAM-2, LAM-3, LAM-4. In the cell-based reporter system for the high throughput screening, LAM-4(11.2μg/ml) could inhibit luciferase activity depending on NF-κB by77percent. The similar results from westenblot also showed that LAM-4(2.8μg/ml,5.6μg/ml and11.2ug/ml) inhibited constitutive and TNF-a-induced phosphorylation of p65. LAM-4inhibited the proliferation of HepG2cell and Hep3B cell with IC5016.5and13.3μg/ml in MTT assay. Results from westenblot indicated that LAM-4down-regulated the expression of c-myc and Survivin in HepG2cells. Flow cytometry and westenblot assay were used to study the effects of extracts of LAM on cell cycle in HepG2cells, and the results showed that LAM could arrest cell cycle of HepG2cells in G0/G1boundary. Inaddition, LAM inhibited the expression of cyclin D1, CDK4, and CDK6, but promoted the expression of p21.Collectively, our study demonstrated that LAM-4, constituent of Lingjia Anticancer Medicine, suppressed the prolifferation of tumor cells by inhibiting the expression of NF-κB regulated genes. The present data preliminary explained the mechanism involved in the antitumor effects of Lingjia Anticancer Medicine and layed the groundwork for further exploration of its effective constituent and molecular mechanism.