SCE administration resulted in observable apoptotic processes, including nuclear pyknosis, enhanced staining intensity, and nuclear fragmentation, in both susceptible and resistant cell lines, as indicated by DAPI staining. In addition, the proportion of apoptotic cells in sensitive/resistant cell lines was substantially elevated, as assessed by double staining flow cytometry, after administration of SCE. Moreover, Western blot analysis of the treated breast cancer cell lines demonstrated a significant reduction in caspase-3, caspase-9, and Bcl-2 protein levels, along with a significant increase in Bax protein expression after SCE administration. With regard to SCE, it could potentially lead to higher counts of positive fluorescent spots after MDC staining and yellow fluorescent spots after GFP-LC3B-mCherry transfection, and result in an augmented expression of autophagy-related proteins such as LC3B, p62, and Beclin-1 in breast cancer cells. Synthesizing the information, SCE could potentially play a role in reversing multidrug resistance in breast cancer cells by blocking their cell cycle, hindering their autophagic pathways, and ultimately interfering with their ability to resist apoptosis.
The objective of this investigation is to uncover the mode of action of Yanghe Decoction (YHD) on subcutaneous tumors that metastasize to the lungs in breast cancer patients, thereby potentially establishing a framework for utilizing YHD in treating breast cancer. Information regarding the chemical compounds within YHD's medicinals, and the targets that these compounds interact with, was retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and SwissTargetPrediction. Targets associated with diseases were sought from GeneCards and Online Mendelian Inheritance in Man (OMIM). Excel's capabilities were employed to pinpoint the common targets and generate a corresponding Venn diagram. A framework depicting protein-protein interactions was created. Employing the R language, Gene Ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were carried out. A total of 53 female SPF Bablc/6 mice were divided into four groups: normal (8 mice), model (15 mice), low-dose YHD (15 mice), and high-dose YHD (15 mice). All groups were treated with the same volume of normal saline, apart from the YHD groups that received escalating doses of YHD through intraperitoneal injections over 30 days. Daily measurements were made of body weight and the dimensions of the tumor. Graphs depicting the relationship between body weight fluctuations and in situ tumor growth were constructed. Following the conclusion of the process, the subcutaneous tumor specimen was collected and examined with hematoxylin and eosin (H&E) staining. By means of PCR and Western blotting, the mRNA and protein levels of hypoxia inducible factor-1 (HIF-1), pyruvate kinase M2 (PKM2), lactate dehydrogenase A (LDHA), and glucose transporter type 1 (GLUT1) were assessed. Out of the total components, 213 active elements from YHD and 185 disease targets were selected for screening. It was hypothesized that YHD might control glycolysis through the HIF-1 signaling pathway, thus influencing breast cancer progression. The animal trials demonstrated that mRNA and protein levels for HIF-1, PKM2, LDHA, and GLUT1 were lower in the high- and low-dose YHD groups compared to the model group. YHD's inhibitory effect on subcutaneous tumor growth in early-stage breast cancer pulmonary metastasis may be attributed to its modulation of glycolysis through the HIF-1 signaling pathway, potentially providing a mechanism to interfere with the development of pulmonary metastasis from breast cancer.
The present investigation explored the molecular underpinnings of acteoside's antitumor effects against hepatoma 22(H22) in mice, with a specific focus on the c-Jun N-terminal kinase (JNK) signaling pathway. Employing subcutaneous inoculation of H22 cells in 50 male BALB/c mice, the resultant mice were categorized into five groups: a model group, low-dose, medium-dose, and high-dose acteoside groups, and a cisplatin group. Consisting of five consecutive days per week, the administration lasted for two weeks for each group. Each group of mice was monitored for general conditions, encompassing mental state, diet, water intake, activity levels, and fur characteristics. The impact on body weight, tumor volume, tumor weight, and the rate of tumor inhibition was assessed and compared in a study that spanned both pre- and post-administration periods. Morphological changes in liver cancer tissues were observed using hematoxylin and eosin (HE) staining, and the expression levels of p-JNK, JNK, Bcl-2, Beclin-1, and LC3 were quantified in each tissue via immunohistochemical and Western blot analysis. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to ascertain the messenger RNA expression levels of JNK, Bcl-2, Beclin-1, and LC3. Innate mucosal immunity Mice in the model and low-dose acteoside treatment groups experienced poor general health, in contrast to the enhanced general well-being noted in the other three treatment groups. Mice in the medium-dose acteoside, high-dose acteoside, and cisplatin groups exhibited a lower body weight compared to the model group, a difference deemed statistically significant (P<0.001). A comparative analysis of tumor volume across the model group and the low-dose acteoside group revealed no statistically significant difference, and the cisplatin group's volume showed no statistically substantial variation from that of the high-dose acteoside group. Compared to the model group, the tumor volume and weight were markedly reduced in the medium-dose acteoside, high-dose acteoside, and cisplatin treatment groups, demonstrating a statistically significant difference (P < 0.0001). The respective tumor-inhibition rates for the low-dose, medium-dose, and high-dose acteoside groups, and the cisplatin group, were 1072%, 4032%, 5379%, and 5644%. A gradual decrease in hepatoma cell counts, observed by HE staining, was correlated with a growing sign of cell necrosis in the acteoside and cisplatin treatment groups. The necrosis was particularly prominent in the high-dose acteoside and cisplatin groups. Samples treated with acteoside and cisplatin displayed an upregulated expression of Beclin-1, LC3, p-JNK, and JNK, as evidenced by immunohistochemical analysis (P<0.05). According to the results of immunohistochemistry, Western blot, and qRT-PCR, Bcl-2 expression was reduced in the medium-dose and high-dose acteoside treatment groups and the cisplatin group (P<0.001). In acteoside and cisplatin treatment groups, Western blot analysis indicated an upregulation of Beclin-1, LC3, and p-JNK (P<0.001). The expression of JNK remained consistent across all groups. qRT-PCR analysis demonstrated an increase in Beclin-1 and LC3 mRNA levels for both acteoside and cisplatin treatment groups (P<0.05). JNK mRNA levels showed a significant increase in the medium- and high-dose acteoside groups and in the cisplatin group (P<0.0001). Acteoside enhances the JNK signaling pathway, which consequently drives apoptosis and autophagy in H22 mouse hepatoma cells, resulting in reduced tumor growth.
Our research delved into how decursin impacted the proliferation, apoptosis, and migration of HT29 and HCT116 colorectal cancer cells, utilizing the PI3K/Akt pathway as a key mechanism. Decursin, at the specified concentrations of 10, 30, 60, and 90 mol/L, was used to treat the HT29 and HCT116 cell lines. Cell counting kit-8 (CCK8), cloning efficiency, Ki67 immunofluorescence, flow cytometry, wound healing, and Transwell assays were used to examine, respectively, the survival, colony formation, proliferation, apoptosis, wound healing, and migration of HT29 and HCT116 cells treated with decursin. A Western blot analysis was conducted to determine the levels of expression of epithelial cadherin (E-cadherin), neural cadherin (N-cadherin), vimentin, B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2-associated X protein (Bax), tumor suppressor protein p53, PI3K, and Akt. diabetic foot infection In comparison to the control group, decursin demonstrably hampered the proliferation and colony count while encouraging the apoptosis of HT29 and HCT116 cells. Furthermore, it noticeably decreased Bcl-2 expression and increased Bax expression. The inhibitory effects of decursin on wound healing and cell migration were pronounced, culminating in a substantial downregulation of N-cadherin and vimentin, and a concomitant upregulation of E-cadherin. This process also entailed a substantial decrease in the expression of PI3K and Akt, along with an increase in the expression of p53. Ultimately, decursin's activity is related to the regulation of epithelial-mesenchymal transition (EMT) via the PI3K/Akt signaling pathway, causing a shift in colorectal cancer cell proliferation, apoptosis, and migration.
The impact of anemoside B4 (B4) on fatty acid metabolism in mice with colitis-associated cancer (CAC) was the focus of this research. Mice underwent treatment with azoxymethane (AOM) and dextran sodium sulfate (DSS) for the creation of the CAC model. Following random allocation, mice were divided into a normal control group, a model group, and groups receiving low-, medium-, and high-dose treatments of anemoside B4. MK-8776 Following the experiment, the length of the mouse colon and the size of the tumor were documented, and hematoxylin-eosin (H&E) staining facilitated the visualization of any pathological alterations present in the colon. The goal of obtaining colon tumor slices was to perform spatial metabolome analysis, specifically evaluating the distribution of compounds related to fatty acid metabolism within the tumor. Using real-time quantitative PCR (RT-qPCR), the mRNA concentrations of SREBP-1, FAS, ACC, SCD-1, PPAR, ACOX, UCP-2, and CPT-1 were ascertained. The model group's body weight (P<0.005) and colon length (P<0.0001) were decreased, and the number of tumors and the pathological score (P<0.001) were elevated, as revealed by the results. The spatial metabolome profile of colon tumors exhibited heightened levels of fatty acids, their associated compounds, carnitine, and phospholipids. The RT-qPCR assay indicated substantial increases (P<0.005, P<0.0001) in the mRNA expression of genes associated with fatty acid de novo synthesis and oxidation, exemplified by SREBP-1, FASN, ACC, SCD-1, ACOX, UCP-2, and CPT-1.