wogonin and Lung-Neoplasms

Wogonin, a natural flavonoid compound, represses cancer cell growth and induces cancer cell apoptosis in diverse malignancies. However, the function of Wogonin in lung cancer cells and its regulatory mechanism deserve to be identified.. A549 and H460 cells were treated with Wogonin, and the cell growth, apoptosis, migration and invasion were measured by CCK-8 and EdU, flow cytometry and Transwell assays. The targeted genes of Wogonin and lung cancer were identified from the TCMSP and Genecards databases, respectively. The STRING database and Cytoscape software were used to establish a PPI network and screen hub genes. GO and KEGG analysis was conducted to explore the functions and signal pathways related to the hub genes. MMP1 expression in lung cancer was analyzed using the UALCAN databases, and GSEA was performed utilizing LinkedOmics. Gelatin zymography assay was used to detect MMP1 activity. MMP1 mRNA expression was detected by qRT-PCR. Besides, MMP1, p-AKT and c-Myc protein were detected by Western Blot assay.. Wogonin could suppress the proliferation, migration and invasion of A549 and H460 cells and induce apoptosis. GO and KEGG enrichment analysis revealed the hub genes were mostly enriched in re-entry into the mitotic cell cycle and apoptosis. The expression of MMP1 was markedly upregulated in lung squamous cell carcinoma, lung adenocarcinoma tissues, and lung cancer cell lines. Wogonin could significantly inhibit MMP1 expression and activity, and overexpression of MMP1 significantly reversed the effect of Wogonin on the malignant phenotypes of A549 and H460 cells. Wogonin inhibited the expression of p-AKT and c-Myc protein by regulating MMP1.. Wogonin can repress lung cancer cells' growth and metastatic potential and promote cell apoptosis via repressing MMP1 expression and modulating PI3K/AKT signaling pathway.

Topics: Apoptosis; Cell Line, Tumor; Cell Movement; Cell Proliferation; Humans; Lung Neoplasms; Matrix Metalloproteinase 1; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction

Lung cancer has a very high incidence rate and is one of the commonly diagnosed tumors in developed countries.. To investigate the effect of wogonin on A549 and A427 lung cancer cells and explore the mechanism involved.. Cell study.. The cytotoxicity effect of wogonin on A549 and A427 lung cancer and BEAS-2B cells was assessed by MTT assay. The onset of apoptosis was assessed by flow cytometry using Annexin V FITC/PI staining. Western blotting was used for the determination of changes in apoptotic protein expression.. Wogonin treatment exhibited cytotoxicity effect selectively on A549 and A427 cells without affecting BEAS-2B normal lung cells. The viability of A549 and A427 cells was reduced to 31% and 34%, respectively, on treatment with 50 μM of wogonin; however, there was no significant reduction in BEAS-2B cell viability on treatment with the same concentration of it. Moreover, the percentage of apoptotic A427 cells showed a significant (p<0.049) increase on treatment with wogonin. Furthermore, the treatment led to a marked increase in the activation of caspase 3/8/9 and the generation of reactive oxygen species (ROS) at 72 h in A427 cells. Digital tomosynthesis studies showed a marked reduction in tumor development on treatment with wogonin.. Wogonin treatment specifically exhibits a cytotoxic effect on lung cancer cells and this effect is associated with activation of apoptosis and generation of reactive oxygen species.

Topics: A549 Cells; Antineoplastic Agents; Apoptosis; Cell Proliferation; China; Drugs, Chinese Herbal; Flavanones; Humans; Lung Neoplasms; Reactive Oxygen Species

Non-small cell lung cancer (NSCLC) is the leading cause of cancer death in the world. Inhibitor of differentiation 1 (Id1) is overexpressed in NSCLC and involved in promoting its progression and metastasis. Identifying natural compounds targeting Id1 may have utility in NSCLC treatment. Here, we sought to determine whether the anti-tumor activities of

Topics: A549 Cells; alpha7 Nicotinic Acetylcholine Receptor; Animals; Antineoplastic Agents; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Flavanones; Flavonoids; Guanosine Triphosphate; Humans; Inhibitor of Differentiation Protein 1; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Invasiveness; Neoplasm Metastasis; Phenotype; Phosphorylation; Plant Extracts; Scutellaria; Shelterin Complex; Telomere-Binding Proteins

A number of significant studies in the field of cell biology have revealed another pattern of intracellular signal transduction in which cells transmit information through the dynamics of key signaling molecules. Dynamical properties of p53 have been demonstrated to be the key factor in dictating cell fate, including cell cycle arrest, permanent cell cycle arrest, and cell death. Previous studies showed a negative feedback regulation pathway between SGK1 and p53, but the dynamics of SGK1 have never been reported before. Therefore, we used different dosing strategies of Wogonin to affect SGK1 dynamics and investigate its impact on cell response. Key factors, such as APAF1, BAX, GADD45A, p21, PML, and YPEL3, which are related to cell cycle arrest, senescence, and apoptosis, were measured at different time points after incubation with Wogonin. Western blot and quantitative reverse transcriptase-polymerase chain reaction analysis were used to examine protein and mRNA expression of these genes. In addition, we also used β-galactosidase staining and flow cytometric analysis to further verify the results. It was found that Wogonin inhibited cell viability and downregulated SGK1 protein levels; 20 μM Wogonin could induce non-small-cell lung cancer A549 cells into cell cycle arrest/senescence/apoptosis after 0.5/2/4 h, respectively; and SGK1 dynamics showed significant differences under different cell responses. Together, our findings showed that SGK1 protein dynamics can be an important part of intracellular signaling, directly influencing cellular response decisions.

Topics: A549 Cells; Apoptosis; Apoptotic Protease-Activating Factor 1; bcl-2-Associated X Protein; Blotting, Western; Carcinoma, Non-Small-Cell Lung; Cell Cycle Checkpoints; Cell Cycle Proteins; Cell Survival; Cellular Senescence; Dose-Response Relationship, Drug; Down-Regulation; Flavanones; Gene Expression Regulation, Neoplastic; Humans; Immediate-Early Proteins; Kinetics; Lung Neoplasms; Molecular Structure; Nuclear Proteins; Protein Serine-Threonine Kinases; Reverse Transcriptase Polymerase Chain Reaction; Time Factors

Increasing evidence from various clinical and experimental studies has demonstrated that the inflammatory microenvironment facilitates tumor metastasis. Clinically, it will be a promising choice to suppress tumor metastasis by targeting inflammatory microenvironment. Our previous studies have demonstrated that wogonin (a bioflavonoid isolated from the traditional Chinese medicine of Huang-Qin) possesses the anti-metastatic and anti-inflammatory activity, but we have little idea about its efficacy on inflammatory-induced tumor metastasis and the mechanism underlying it. In this study, we focused on epithelial mesenchymal transition (EMT), the first step of tumor metastasis, to evaluate the effects of wogonin on tumor metastasis in inflammatory microenvironment. We found that wogonin inhibited THP-1 conditioned-medium- (CM-) and IL-6-induced EMT by inactivating STAT3 signal. And in wogonin-treated A549 cells which pretreated with THP-1 CM or IL-6, the expression level of E-cadherin, an EMT negative biomarker, increased while that of N-cadherin, Vimentin, and EMT-related transcription factors including Snail and Twist decreased. Moreover, wogonin inhibited IL-6-induced phosphorylation of STAT3, prevented p-STAT3 dimer translocation into the nucleus, and suppressed the DNA-binding activity of p-STAT3. Interestingly, similar results were obtained in the tumor xenografts mice, including downregulation of p-STAT3, N-cadherin, and Vimentin while up-regulation of E-cadherin. Wogonin also inhibit the metastasis of A549 cells in vivo. Taken all data together, we concluded that wogonin suppresses tumor cells migration in inflammatory microenvironment by inactivating STAT3 signal.

Topics: Adenocarcinoma; Animals; Drugs, Chinese Herbal; Epithelial-Mesenchymal Transition; Flavanones; Humans; Inflammation; Interleukin-6; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Signal Transduction; STAT3 Transcription Factor; Tumor Microenvironment

The objective of the present study was to investigate the therapeutic efficacy of flavonoid components in Scutellaria baicalensis on proliferation, metastasis and lung cancer-associated inflammation during nicotine induction in the A549 and H1299 lung cancer cell lines. After experimental period, augmentation of proliferation was observed, accompanied by marked decrease in apoptotic cells in nicotine-induced lung cancer cells; additionally, nicotine-exposed cells exhibited increased invasive and migratory abilities based on invasion and wound-healing assay. Flavones in Scutellaria, baicalin, baicalein and wogonin significantly counteracted the above deleterious changes. Moreover, assessment of tumor apoptotic and metastatic factors on mRNA levels by quantitative PCR and protein levels by western blotting revealed that these phytochemical treatments effectively negated nicotine-induced upregulated expression of bcl-2, bcl-2/bax ratio, caspase-3, matrix metalloproteinase (MMP)-2 and MMP-9 as well as downregulated expression of bax. Further analysis of inflammatory markers such as tumor necrosis factor (TNF)-α and interleukin (IL)-6 in cell culture supernatant and mRNA and protein expression of nuclear transcription factor-kappaB (NF-κB) and I kappa B-alpha (IκB-α) was carried out to substantiate the anti-inflammatory effect of flavones in Scutellaria in nicotine-exposed lung cancer cells. The therapeutic effects observed in the present study are attributed to the potent potential against proliferation, metastasis and inflammatory microenvironment by flavonoid components in Scutellaria in nicotine-induced lung cancer cells.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Cell Line, Tumor; Cell Proliferation; Flavanones; Flavonoids; Gene Expression Regulation, Neoplastic; Humans; In Vitro Techniques; Inflammation; Lung Neoplasms; Neoplasm Metastasis; Nicotine; Plant Extracts; Scutellaria baicalensis

Wogonin is a plant monoflavonoid which has been reported to inhibit cell growth and/or induce apoptosis in various tumors. The present study examined the apoptosis-inducing activity and underlying mechanism of action of wogonin in A549 cells. The results showed that wogonin was a potent inhibitor of the viability of A549 cells. Apoptotic protein changes detected after exposure to wogonin included decreased XIAP and Mcl-1 expression, increased cleaved-PARP expression and increased release of AIF and cytochrome C. Western blot analysis showed that the activity of c-Myc/Skp2 and HDAC1/HDAC2 pathways, which play important roles in tumor progress, was decreased. Quantitative PCR identified increased levels of c-Myc mRNA and decreased levels of its protein. Protein levels of Fbw7α, GSK3β and Thr58-Myc, which are involved in c-Myc ubiquitin-dependent degradation, were also analyzed. After exposure to wogonin, Fbw7α and GSK3β expression decreased and Thr58-Myc expression increased. However, MG132 was unable to prevent c-Myc degradation. The present results suggest that wogonin has multiple anti-cancer effects associated with degradation of c-Myc, SKP2, HDAC1 and HDAC2. Its ability to induce apoptosis independently of Fbw7α suggests a possible use in drug-resistance cancer related to Fbw7 deficiency. Further studies are needed to determine which pathways are related to c-Myc and Fbw7α reversal and whether Thr58 phosphorylation of c-Myc is dependent on GSK3β.

Topics: Adenocarcinoma; Adenocarcinoma of Lung; Apoptosis; Cell Cycle; Cell Cycle Proteins; Cell Line, Tumor; Cell Proliferation; Cell Survival; F-Box Proteins; F-Box-WD Repeat-Containing Protein 7; Flavanones; Gene Expression Regulation, Neoplastic; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Histone Deacetylase 1; Histone Deacetylase 2; Humans; Lung Neoplasms; Membrane Potential, Mitochondrial; Proto-Oncogene Proteins c-myc; RNA, Messenger; S-Phase Kinase-Associated Proteins; Signal Transduction; Ubiquitin-Protein Ligases

Combination of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) with other agents is a promising strategy to overcome TRAIL resistance in malignant cells. Wogonin, a flavonoid originated from Scutellaria baicalensis Georgi, has been shown to enhance TRAIL-induced apoptosis in malignant cells in in vitro studies. However, whether wogonin enhances TRAIL's antitumor activity in vivo has never been studied. In this study, the effect of combination of TRAIL and wogonin was tested in a non-small-cell lung cancer xenografted tumor model in nude mice. Consistent with the in vitro study showing that wogonin sensitized A549 cells to TRAIL-induced apoptosis, wogonin greatly enhanced TRAIL-induced suppression of tumor growth, accompanied with increased apoptosis in tumor tissues as determined by TUNEL assay. The expression levels of antiapoptotic proteins including long form of cellular FLICE-like inhibitory protein (cFLIPL), X-linked inhibitor of apoptosis protein (XIAP), and cellular inhibitor of apoptosis protein 1 and 2 (cIAP-1 and cIAP-2) were markedly reduced in both cultured cells and xenografted tumor tissues after co-treatment with wogonin and TRAIL. The down-regulation of these antiapoptotic proteins was likely mediated by proteasomal degradation that involved intracellular reactive oxygen species (ROS), because wogonin robustly induced ROS accumulation and ROS scavengers butylated hydroxyanisole (BHA) and N-acetyl-L-cysteine (NAC) and the proteasome inhibitor MG132 restored the expression of these antiapoptotic proteins in cells co-treated with wogonin and TRAIL. These results show for the first time that wogonin enhances TRAIL's antitumor activity in vivo, suggesting this strategy has an application potential for clinical anticancer therapy.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Baculoviral IAP Repeat-Containing 3 Protein; Carcinoma, Non-Small-Cell Lung; CASP8 and FADD-Like Apoptosis Regulating Protein; Cell Line, Tumor; Drug Synergism; Flavanones; Gene Expression Regulation, Neoplastic; Humans; Inhibitor of Apoptosis Proteins; Lung Neoplasms; Male; Mice; Mice, Nude; Neoplasm Transplantation; Reactive Oxygen Species; Signal Transduction; TNF-Related Apoptosis-Inducing Ligand; Ubiquitin-Protein Ligases

Scutellaria baicalensis root is widely used in China as an adjuvant to orthodox chemotherapy of lung cancer. However, functional biomarkers of this plant for anti-lung cancer activity have not yet been reported. We therefore determined the growth inhibition activity by MTT assay of eight solvent extracts of S. baicalensis in the human lung cancer cell line SK-MES-1. This activity was then mapped onto the secondary metabolite profile of crude extracts by principal components analysis (PCA) of proton NMR and HPLC-UV data. NMR- and HPLC-PCA maps revealed highest inhibitory activity for the non-aqueous extracts. The first two components of both maps discriminated extract activity mainly based on the differential content of three compounds, which were then tested individually. The IC(50) values for baicalin (IC(50): 64+/-5 microM), baicalein (IC(50): 80+/-6 microM) and wogonin (IC(50): 39+/-10 microM) were comparable to that of the antineoplastic cisplatin (IC(50): 79+/-16 microM). A partial least squares regression (PLS)-NMR model highly correlated with the corresponding PLS-HPLC model for prediction of inhibition. Secondary metabolite mapping of lung cancer growth inhibitors in crude extracts may be an important first step to qualify Chinese herbal prescriptions required for meaningful clinical trials of such integrated therapies.

Topics: Antineoplastic Agents, Phytogenic; Cell Line, Tumor; Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Humans; Least-Squares Analysis; Lung Neoplasms; Magnetic Resonance Spectroscopy; Plant Extracts; Principal Component Analysis; Scutellaria

Wogonin, a naturally occurring plant flavonoid, is isolated from Chinese herbal plants Scutellaria baicalensis Georgi and S. barbata D. Don. The extract of S. baicalensis Georgi has been added to an assortment of health drinks or food supplements. Wogonin has been reported to exhibit anticancer and anti-inflammatory properties. Cyclooxygenase-2 (COX-2) is a key enzyme in the production of prostaglandins in inflammatory conditions. In this study, the effect of wogonin on phorbol 12-myristate 13-acetate (PMA)-induced COX-2 expression was investigated. It showed that wogonin inhibited PMA-induced COX-2 protein and mRNA levels in human lung epithelial cancer cells, and the mechanism of this inhibition was at the transcriptional level by using COX-2 gene promoter assay. Among various signal inhibitors, the mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 also inhibited PMA-induced COX-2 expression and COX-2 promoter activation. The activity of AP-1-driven promoter, but not nuclear factor-kappa B (NF-kappaB), was inhibited by U0126. The data indicated that MEK1/2-AP-1 is very important for PMA-induced COX-2 expression. Wogonin also inhibited PMA-induced AP-1 activation and the expression of c-Jun, a key component of AP-1. Taken together, it is suggested that wogonin inhibits PMA-induced COX-2 gene expression by inhibiting c-Jun expression and AP-1 activation in A549 cells.

Topics: Anti-Inflammatory Agents; Beverages; Cell Line, Tumor; Cell Survival; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Epithelial Cells; Flavanones; Gene Expression Regulation, Enzymologic; Humans; Lung Neoplasms; Promoter Regions, Genetic; Proto-Oncogene Proteins c-jun; Tetradecanoylphorbol Acetate; Transcription Factor AP-1

Dihydrodiol dehydrogenase (DDH) is a member of the aldo-keto reductases superfamily (AKR1C1-AKR1C4), which plays central roles in the metabolism of steroid hormone, prostaglandin and xenobiotics. We have previously detected overexpression of DDH as an indicator of poor prognosis and chemoresistance in human non-small lung cancer (NSCLC). We also found DDH expression to be closely related to chronic inflammatory conditions. The aim of this study was to investigate the links between inflammation, DDH expression and drug resistance in NSCLC cells. We showed that pro-inflammatory mediators including interleukin-6 (IL-6) could induce AKR1C1/1C2 expression in NSCLC cells and increase cellular resistance to cisplatin and adriamycin. This effect was nullified by Safingol, a protein kinase C inhibitor. Moreover, the expression of AKR1C1/1C2 was inversely correlated to NBS1 and apoptosis-inducing factor (AIF). We also showed that IL-6-induced AKR1C1/1C2 expression and drug resistance were inhibited by wogonin and chrysin, which are major flavonoids in Scutellaria baicalensis, a widely used traditional Chinese and Japanese medicine. In conclusion, this study demonstrated novel links of pro-inflammatory signals, AKR1C1/1C2 expression and drug resistance in NSCLC. The protein kinase C pathway may play an important role in this process. Overexpression of AKR1C1/1C2 may serve as a marker of chemoresistance. Further studies are warranted to evaluate wogonin and chrysin as a potential adjuvant therapy for drug-resistant NSCLC, especially for those with AKR1C1/1C2 overexpression.

Topics: 20-Hydroxysteroid Dehydrogenases; Apoptosis; Carcinoma, Non-Small-Cell Lung; Cell Cycle; Cell Line, Tumor; DNA Repair; Dose-Response Relationship, Drug; Doxorubicin; Drug Resistance, Neoplasm; Flavanones; Flavonoids; Humans; Hydroxysteroid Dehydrogenases; Interleukin-6; Lung Neoplasms