Exploring the mechanism and experimental verification of puerarin in the treatment of endometrial carcinoma based on network pharmacology and bioinformatics analysis

Lin et al. BMC Complementary Medicine and Therapies https://doi.org/10.1186/s12906-022-03623-z BMC Complementary Medicine and Therapies (2022) 22:150 Open Access RESEARCH Exploring the mechanism and experimental verification of puerarin in the treatment of endometrial carcinoma based on network pharmacology and bioinformatics analysis Zhiheng Lin1†, Xiaohui Sui1†, Wenjian Jiao1, Ying Wang1 and Junde Zhao1,2* Abstract Endometrial carcinoma is one of the two cancers with rising mortality and morbidity in recent years. In the light of many controversies about its treatment, it is urgent to construct a new prognostic model and to find out new therapeutic directions. As a small drug molecule widely used in clinical treatment and experimental research in China, puerarin has recently been proven to have obvious anti-cancer effects in multiple cancer cells. In this study, bioinformatics analysis and experimental validation were used to explore the potential mechanism of puerarin for endometrial carcinoma and construct a prognostic model. A total of 22 drug-related differential genes were found by constructing a database of drug targets and disease genes. The protein–protein interaction network was constructed for GO and KEGG enrichment analysis to initially explore the potential mechanism of its therapeutic effects. To construct the prognostic model, validation was performed by risk regression analysis and LASSO analysis. Finally, two prognostic genes—PIM1 and BIRC5 were determined to establish high and low risk groups. Kaplan–Meier analysis displayed a higher survival rate in the low-risk group than in the high-risk group. ROC curves indicated the stable and good effect in prediction (one-year AUC is 0.626; two-year AUC is 0.620; three-year AUC is 0.623). The interrelationship between immunity and its disease was explored by immune infiltration analysis. Finally, the potential effect of puerarin on endometrial carcinoma cells was further verified by experiments. Keywords: Puerarin, UCEC, Network Pharmacology, Bioinformatics Analysis Introduction Endometrial carcinoma is the most common tumor in female genital tumors, occurring mostly in menopausal women aged 60 + on average [1]. According to the American Cancer Association, about 66,570 new endometrial carcinoma cases were diagnosed in the United States in † Zhiheng Lin and Xiaohui Sui contribute equally to this work and co-first authors. *Correspondence: 1 Shandong University of Traditional Chinese Medicine, Jinan 250014, Shandong, China Full list of author information is available at the end of the article 2021, with 130,000 women who died of endometrial carcinoma. Endometrial carcinoma is one of the two cancers with rising mortality and morbidity in recent years [2], which seriously affects the quality of life and the health of patients. Endometrial carcinoma occurrence is associated with excessive estrogen secretion [3]. Endometrial carcinoma is an estrogen-dependent tumor, accounting for 80% of newly diagnosed endometrial carcinoma cases. The risk of disease increases with the duration of estrogen use and remains years after estrogen withdrawal [4, 5]. At present, the endometrial carcinoma is mainly subject to surgical treatment, such as total hysterectomy, bilateral tubal ovariectomy, paraaortic and pelvic © The Author(s) 2022. 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The Creative Commons Public Domain Dedication waiver (http://creativeco mmons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Lin et al. BMC Complementary Medicine and Therapies (2022) 22:150 Page 2 of 15 lymphadenectomy. It can also be treated through adjuvant therapies such as radiotherapy, chemotherapy and hormonal therapy [6]. There are still many controversies in the treatment of endometrial carcinoma, including the evaluation of lymph and the selection of adjuvant treatment. There is relatively little space for treatment options for advanced as well as metastatic tumors [7]. Considering the controversial and limited treatment methods of endometrial carcinoma, it is particularly important to explore the small molecule drugs for treating endometrial cancer, and to establish a new development model to accurately and conveniently predict the survival cycle of endometrial carcinoma patients. Studies show that traditional Chinese medicine has an overall macro-control effect on disease, and has an obvious curative effect on disease control and prevention [8]. With their potential tumor selectivity and low cytotoxicity, many natural small-molecule substances present in TCM have attracted wide attention in the research and development of tumor drugs [9]. As a bioactive small molecule of isoflavone glycoside extracted from pueraria lobata, puerarin has bio-functional activities such as promoting bone formation, protecting cardiomyocytes, protecting nerve function, anti-inflammatory and alleviating pain [10–15]. Puerarin has been widely used in experimental research and clinical treatment. Puerarin injections made from puerin have been widely used in China [16]. More studies have shown that puerarin also shows significant anticancer effects in multiple cancer cell lines, such as prostate, bladder, colon, breast, and cervical cancer [17–21]. However, puerarin has not been intensively studied in endometrial carcinoma treatment. In this context, network pharmacology and Bioinformatics Analysis were used to to explore the intrinsic mechanism of puerarin in the treatment of endometrial carcinoma, establish the prognostic model of drug-related differential gene, and validate it with cellular experiments. tumor group samples were obtained. Data were extracted and standardized using R language (R 4.1.1). Differential genes between normal and tumor samples were screened using the ‘limma’ R package. Screen parameter setting: P < 0.05,PDR < 0.05, ∣LOGFC∣ > 1. Differentially expressed genes (DEGs) for endometrial carcinoma were obtained. Materials and methods Obtain the dataset of drug target genes First, 55 drug targets related to puerarin pharmaceutical molecules were sea (...truncated)