Prediction of the therapeutic effect of total flavonoids from Kunlun Snow Chrysanthemum on non-small cell lung cancer using network pharmacology technology and preliminary validation study
Lung cancer is one of the most common cancers worldwide. About 85% of lung cancer is non-small cell lung cancer (NSCLC). The main clinical treatments for lung cancer include surgery, chemotherapy, radiation therapy, and biological therapy. Although complete surgical resection is effective, the prerequisite is that the disease is medically operable and can be appropriately staged. Chemotherapy drugs have non-specific properties and often produce serious toxic side effects. Traditional Chinese medicine has played a unique role in the prevention and treatment of tumors due to its multi-target, multi pathway, and multi effect properties. Several traditional Chinese medicine ingredients and their extracts have been found to have good anti-tumor effects, such as matrine, genistein, paclitaxel, etc., some of which have been widely used in clinical practice. Kunlun Snow Chrysanthemum is also known as Coreopsis tinctoria Nutt. Modern pharmacological studies have shown that Xinjiang Kunlun Snow Chrysanthemum is rich in various chemical components such as polyphenols, flavonoids, and polysaccharides. Among them, flavonoids have important pharmacological effects such as lowering blood lipids, regulating blood sugar, and anti-tumor effects. However, due to the abundance of flavonoids, multiple targets, and multiple pathways of action, there are certain difficulties in studying their molecular mechanisms for treating diseases.
Network pharmacology is a research method based on systems biology and multidisciplinary pharmacology, which constructs a multi-level interaction network of “drug targets disease” to analyze drug targets and mechanisms at the overall level. This article uses network pharmacology combined with in vitro cell experiments to analyze the anti-tumor components and mechanisms of Kunlun chrysanthemum total flavonoids (KCTF) on NSCLC, providing experimental evidence for its subsequent drug development and clinical application.

Flavonoids widely present in various plants are strong antioxidants that can effectively eliminate free radicals in the body and have important pharmacological effects such as anti-tumor, blood pressure lowering, and anti-aging. This study analyzed the total flavonoids in Kunlun Snow Chrysanthemum using HPLC technology. The results showed that the relatively high content of flavonoids included epigallocatechin gallate, quercetin, luteolin, catechins, etc. Numerous research results have shown that flavonoids such as quercetin and luteolin have good anti-tumor activity against various solid tumors. In vitro cell experiments have shown that total flavonoids from Kunlun Snow Chrysanthemum have a significant inhibitory effect on the proliferation of human colon cancer HCTI16 cells and Caco-2 cells. However, the characteristics of traditional Chinese medicine’s multi-component, multi-target, and multi pathway synergistic effects make modern research on its mechanism of action difficult.
In 2007, Hopkins proposed network pharmacology, which represents the relationship between drugs, targets, and diseases in a network. The use of network pharmacology can systematically analyze the components of traditional Chinese medicine, explore the common targets of drugs and diseases, and further analyze biological functions and signaling pathways. This holistic and systematic research model is consistent with the holistic theory of traditional Chinese medicine.
In this study, 68 components of KCTF were screened using TCMSP, and 891 targets of 27 flavonoid components were predicted. After intersecting with NSCLC targets, 217 possible targets for KCTF intervention in NSCLC were obtained. Among them, important targets such as MAPK1, EGFR, Tp53, Akt, Bax, Bcl-2, Caspase 3 are related to multiple chemical components, indicating a close synergistic relationship between the active ingredients of KCTF. To further analyze the mechanism of KCTF intervention in NSCLC, the relationship between the two was analyzed by constructing a PPI network and enriching KEGG biological pathways. The results showed that PI3K Akt signaling pathway, Bax/Bcl-2 anti apoptotic signaling pathway, p53 signaling pathway, MAPK signaling pathway, etc. were related to the anti-tumor effect of KCTF.
The main mechanisms of action of anti-tumor drugs are inducing tumor cell apoptosis, inhibiting the cell cycle, and suppressing migration. Research has shown that many plant flavonoids (such as curcumin, anthocyanins, epigallocatechin gallate, quercetin, etc.) can interfere with the PI3K Akt signaling pathway, directly inhibit PI3k or block the PI3K Akt signaling pathway, further regulate the expression of downstream effector molecules, accelerate tumor cell cycle, promote cell apoptosis, and inhibit cell proliferation, achieving anti-tumor effects. Xiang et al. found that quercetin induces apoptosis in HeLa cells by downregulating the expression of PI3k, Akt, and Bcl-2 in the PI3K Akt signaling pathway, upregulating Bax expression, and causing cell cycle arrest in G0/G. Osmanthus extract can exert anti-tumor activity by inhibiting the expression of MMP-2 and MMP-9 in the PI3K Akt signaling pathway, suppressing the proliferation of human melanoma cells, and inducing their apoptosis. The experimental results at the cellular level of this study showed that total flavonoids from Chrysanthemum morifolium significantly inhibited the proliferation of A549 cells, promoted cell apoptosis, and inhibited cell migration. At the mRNA level, it was verified that total flavonoids from Chrysanthemum morifolium upregulated the expression of Bax and downregulated the expression of Bcl-2 and MMP-2. When Bax is highly expressed in cells, the cells are sensitive to death signals and promote apoptosis. When Bcl-2 is highly expressed, it can form heterodimers with Bax, inhibiting cell apoptosis. Therefore, the ratio of Bax/Bcl-2 in cells plays an important role in determining the sensitivity of cell apoptosis. Therefore, Western blot validation experiments at the cellular level showed that total flavonoids from Scutellaria baicalensis significantly upregulated the ratio of Bax/Bcl-2, promoting apoptosis in A549 cells. Based on the results of network pharmacology analysis, total flavonoids from Scutellaria baicalensis can intervene in NSCLC through multiple targets and pathways, and its anti-tumor activity may be related to the expression of apoptosis genes Bax and Bcl-2.
This article observed the anti-tumor effect of total flavonoids extract from Chrysanthemum morifolium on A549 cells at the cellular level. However, due to the low yield of total flavonoids purified by macroporous resin and the high IC50 of the drug on cell inhibition, the purification method will be optimized for in vivo animal experiments to increase the yield of flavonoids and reduce the drug concentration. Combined with the results of network pharmacology analysis, the molecular mechanism of its anti-tumor effect will be further studied.