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1.
Article in Chinese | WPRIM | ID: wpr-940290

ABSTRACT

ObjectiveProteoglycan TPG-1 isolated from Trametes robiniophila(Huaier) has proved to have anti-hepatoma activity, and this paper aims to explore the molecular mechanism. MethodHuman hepatoma SK-HEP-1 cells were treated with TPG-1 (0, 0.05, 0.1, 0.25, 0.5, 1 g·L-1). Then cell survival was detected by methyl thiazolyl tetrazolium (MTT) and apoptosis by flow cytometry. In addition, expression of genes in SK-HEP-1 cells treated with or without TPG-1 was examined by DNA microarray to preliminarily explore the anti-hepatoma molecular mechanism of TPG-1. ResultTPG-1 inhibited the proliferation of SK-HEP-1 cells as compared with the blank group (P<0.01). After treatment with 1 g·L-1 TPG-1 for 48 h, the apoptosis rate of SK-HEP-1 cells increased (P<0.01), and TPG-1 promoted the cleavage of cysteinyl aspartate specific proteinase (Caspase)-3 and Caspase-7, the key mediators of apoptosis (P<0.01). Additionally, TPG-1 (1 g·L-1) suppressed the migration of SK-HEP-1 cells (P<0.05). A total of 971 differentially expressed genes (DEGs) were identified in SK-HEP-1 cells after treatment with TPG-1, with 486 up-regulated and 485 down-regulated. These DEGs were mainly involved in the Gene Ontology (GO) terms of interleukin-6 (IL-6) biosynthesis, antigen processing and presentation, superoxide dismutase activity, positive regulation of mitogen-activated protein kinase kinase kinase (MAPKKK) cascade, nature killer (NK) cell chemotaxis, and chemokine biosynthesis, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of nucleotide-binding oligomerization domain (NOD)-like receptor signaling pathway, apoptosis, Toll-like receptor signaling pathway, retinoic acid-inducible gene-Ⅰ (RIG-Ⅰ)-like receptor signaling pathway, T-cell receptor signaling pathway, and chemokine signaling pathway. Western blot results showed that TPG-1 (1 g·L-1) activated mitogen-activated protein kinase (MAPK) signaling pathway in SK-HEP-1 cells (P<0.01). ConclusionProteoglycan TPG-1 inhibited the proliferation and migration, and induced apoptosis of human hepatoma SK-HEP-1 cells. Up-regulation of MAPK signaling pathway may be responsible for the growth inhibition of human hepatoma SK-HEP-1 cells by TPG-1.

2.
Article in Chinese | WPRIM | ID: wpr-940291

ABSTRACT

ObjectiveTo investigate effect of aqueous extract of Trametes robiniophila (TRM,Huaier) on autophagy of human prostate cancer VCaP cells and Lamin B1 expression, so as to uncover its role in the proliferation of VCaP cells. MethodThe inhibitory effect of 0, 2, 4, 6, 8, 10 g·L-1 TRM aqueous extract on the proliferation of human prostate cancer VCaP cells at different time points were determined by cell counting kit-8 (CCK-8) assay. Acridine orange staining was conducted for analyzing the effect of TRM aqueous extract on the formation of autolysosomes in VCaP cells. After medication, the expression of microtubule-associated protein Ⅰ light chain 3 (LC3), autophagy-related protein 3 (Atg3), autophagy-related protein 5 (Atg5), and autophagy-related protein 7 (Atg7) in VCaP cells were detected by Western blot. The effect of TRM aqueous extract alone and its combination with autophagy inhibitor bafilomycin A1 on the proliferation of VCaP cells were assayed by CCK-8 assay. RNA interference technology was used to explore the role of Lamin B1 in anti-proliferation of VCaP cells by TRM. ResultCompared with the blank group, TRM aqueous extract inhibited the proliferation of human prostate cancer VCaP cells in a time- and concentration-dependent manner (P<0.01). Acridine orange staining showed that TRM aqueous extract promoted the formation of autolysosomes in VCaP cells. As revealed by Western blotting, TRM aqueous extract up-regulated the expression levels of LC3-Ⅱ, Atg3, Atg5, and Atg7 in contrast to those in the blank group (P<0.05). All these indicated that TRM aqueous extract induced the autophagy of VCaP cells. In addition, autophagy inhibition impaired the sensitivity of VCaP cells to TRM aqueous extract (P<0.05). The comparison with the blank group showed that TRM aqueous extract inhibited Lamin B1 protein expression in VCaP cells (P<0.01), which in turns weakened the sensitivity of VCaP cells to TRM aqueous extract. ConclusionTRM aqueous extract inhibited the proliferation of human prostate cancer VCaP cells possibly by inducing autography and down-regulating Lamin B1 expression. This study has provided a theoretical basis for the clinical application of TRM.

3.
Article in Chinese | WPRIM | ID: wpr-940292

ABSTRACT

ObjectiveTo investigate the effect of Draconis Sanguis petroleum ether fraction (DSPEF) on the proliferation, apoptosis, migration, and autophagy of human gastric cancer HGC-27 and MGC-803 cells, and preliminarily elucidate its molecular mechanism. MethodCell counting kit-8 (CCK-8) assay was used to detect the effect of DSPEF at different concentrations (0, 20, 40, 60, 80 mg·L-1) on the proliferation of HGC-27 and MGC-803 cells after 24, 48, 72 h. Hoechst staining and flow cytometry were used to explore the effects of DSPEF at different concentrations on the apoptosis and apoptosis rate of HGC-27 and MGC-803 cells after 48 h treatment, respectively. The wound healing assay and acridine orange staining were used to investigate the effects of DSPEF on the migration and autophagy of HGC-27 and MGC-803 cells, respectively. Western blot was used to detect the expression levels of signaling pathway-related proteins in HGC-27 and MGC-803 cells treated with DSPEF for 48 h. ResultCompared with the control group, DSPEF(30 mg·L-1) inhibited the proliferation and migration of HGC-27 and MGC-803 cells in a concentration- and time-dependent manner (P<0.05), and induced the apoptosis (P<0.01) and autophagy of HGC-27 and MGC-803 cells. DSPEF (60 mg·L-1) down-regulated the protein levels of phosphorylated mammalian target of rapamycin (p-mTOR) (P<0.05, P<0.01) and down-regulated phospho-signal transducer and activator of transcription 3 (p-STAT3) in HGC-27 and MGC-803 cells (P<0.01), suggesting that DSPEF presumedly inhibited the proliferation and migration of human gastric cancer HGC-27 and MGC-803 cells and induced their apoptosis and autophagy by inhibiting the mTOR/STAT3 signaling pathway. ConclusionThe down-regulation of the mTOR/STAT3 signaling pathway may be involved in the anti-gastric cancer effect of DSPEF. This study is expected to provide a reference for the investigation of the anti-tumor effect of Draconis Sanguis.

4.
Article in Chinese | WPRIM | ID: wpr-940293

ABSTRACT

Malignant tumor is a serious threat to human life and health. The prevalence and mortality of malignancies in China are increasing year by year. Conquering cancer has become a difficult problem for human beings. Chemical drug therapy combined with molecular targeted therapy is a general and preferred anti-tumor clinical scheme, but the side effects and the drug resistance of cancer cells often hinder the efficacy. Therefore, it is of great significance to study the mechanism of drug resistance and the methods to reverse drug resistance. Chinese medicine has the characteristics of complex components, multiple targets, low toxicity, etc. A large number of experimental studies have demonstrated that the effective components or extracts of Chinese medicine can inhibit the proliferation, migration, and invasion of cancer cells, and induce apoptosis, autophagy, differentiation, and senescence. In clinical practice, Chinese medicine has been applied to the protection against ttumor, adjuvant treatment, and later consolidation. The research on Chinese medicine is expected to promote drug resistance reversal and cancer therapy. Studies have shown that the combination of Chinese medicine and chemotherapy can reverse drug resistance and increase efficacy, which has become the mainstream trend of cancer treatment. This study reviewed the mechanisms of the drug resistance of cancer cells induced by self-protective autophagy, gene mutation, high expression of enzymes, abnormal signaling pathways, and abnormal expression of RNA and protein, and summarized how compounds isolated from Chinese medicine, single drug and its extract, and classic anti-cancer prescription reversed the drug resistance to lay a solid foundation for the further investigation of the anti-tumor effect of Chinese medicine.

5.
Article in Chinese | WPRIM | ID: wpr-879127

ABSTRACT

As a traditional Chinese medicine, Chinese dragon's blood has multiple effects, such as activating blood to remove blood stasis, softening and dispelling stagnation, astringent and hemostasis, clearing swelling and relieving pain, regulating menstruation and rectifying the blood, so it is called "an effective medicine of promoting blood circulation". It has been widely used clinically to treat a variety of diseases. With the further research on Chinese dragon's blood, its anti-tumor medicinal value is gradually emerging. Modern pharmacological studies have shown that Chinese dragon's blood exerts anti-tumor effects mainly by inhibiting cell proliferation, inducing apoptosis, inducing DNA damage and cell cycle arrest, inducing senescence and autophagy of tumor cells, inhibiting metastasis and angiogenesis, as well as reversing multidrug resistance. This article focuses on the research progress on anti-tumor effects of Chinese dragon's blood extract and its chemical components, with a view to provide new references for the in-depth research and reasonable utilization of Chinese dragon's blood.


Subject(s)
Female , China , Dracaena , Plant Extracts , Resins, Plant
6.
Article in Chinese | WPRIM | ID: wpr-828395

ABSTRACT

This study aims to investigate the effect of Huaier aqueous extract on the growth and metastasis of human non-small cell lung cancer NCI-H1299 cells and its underlying mechanisms. MTT assay was used to detect the effect of Huaier aqueous extract on the proliferation of NCI-H1299 cells. Flow cytometry was used to examine the effect of Huaier aqueous extract on the apoptosis, cell cycle, and ROS level of NCI-H1299 cells. Wound healing assay was used to evaluate the effect of Huaier aqueous extract on the migration ability of NCI-H1299 cells. Western blot was used to detect the levels of proteins involving apoptosis, epithelial-mesenchymal transition(EMT), and MAPK signaling pathway in NCI-H1299 cells exposed to Huaier aqueous extract. The results showed that Huaier aqueous extract inhibited the proliferation of NCI-H1299 cells, and induced cell-cycle arrest at the phase S. Huaier aqueous extract promoted the apoptosis of NCI-H1299 cells by down-regulating the expression of anti-apoptotic protein Bcl-2. Moreover, Huaier aqueous extract increased ROS level and induced ferroptosis in NCI-H1299 cells. EMT played a critical role in cancer metastasis. Huaier aqueous extract reduced the migration ability of NCI-H1299 cells by inhibiting EMT of NCI-H1299 cells. In addition, this study revealed that Huaier aqueous extract inhibited MAPK signaling pathway in human non-small cell lung cancer NCI-H1299 cells, which may be one of Huaier's mechanisms in inhibiting growth and metastasis of NCI-H1299 cells. This study provides a new theoretical basis for the clinical treatment of lung cancer with Huaier, and important reference significance for further studies on the anti-tumor mechanisms of Huaier.


Subject(s)
Humans , Apoptosis , Carcinoma, Non-Small-Cell Lung , Cell Line, Tumor , Cell Proliferation , Complex Mixtures , Lung Neoplasms , Trametes
7.
Article in Chinese | WPRIM | ID: wpr-862659

ABSTRACT

Objective::To investigate the protective effect of Longxue Tongluo capsule (LTC) on H2O2-induced injury of PC12 cells and its potential mechanism. Method::An in vitro injury model in PC12 cells was established with 500 μmol·L-1 H2O2.The experiment was divided into control group, injury model group (H2O2 500 μmol·L-1), and Longxuetongluo capsule group (LTC, 1, 2, 4 mg·L-1). Cell counting kit-8 (CCK-8) assay, intracellular reactive oxygen species (ROS), mitochondrial membrane potential, apoptosis of PC12 cells, and Western blot were used to evaluate the protective effect of LTC on PC12 cells induced by H2O2. Result::Compared with the control group, the cell viability was significantly decreased in the injury model group (P<0.01), intracellular ROS level was significantly increased (P<0.01), mitochondrial membrane potential was decreased, while apoptosis of PC12 cells was significantly increased (P<0.01), and the expression of cleaved poly adenosine diphospho ribose polymerase (PARP) was also increased significantly (P<0.01). Compared with the injury model group, pretreatment with LTC at the concentrations of 2 and 4 mg·L for 6 h significantly increased cell viability in PC12 cells exposed to H2O2 (P<0.01). Moreover, pretreatment with LTC reduced intracellular ROS level (P<0.05), maintained mitochondrial membrane potential, and inhibited apoptosis of PC12 cells induced by H2O2 in a dose-dependent manner (P<0.01). The results of western blotting showed that pretreatment with LTC significantly reduced the expression of cleaved PARP (P<0.01). Conclusion::LTC exerts a significant protective effect against H2O2-induced PC12 cells injury through inhibition of neuronal apoptosis by suppressing intracellular oxidative stress, maintaining mitochondrial function, and promoting DNA repair.

8.
Article in Chinese | WPRIM | ID: wpr-774537

ABSTRACT

The research of anti-hepatocellular carcinoma(HCC) drug has attracted more and more attention. Natural products are the important source of active compounds for cancer treatment. A biflavonoid HIS-4 was isolated from Resina draconis in our previous study. MTT assay, hoechst staining, and flow cytometry analysis were used to investigate the effects of HIS-4 on the proliferation and apoptosis of human hepatoma HepG2 and SK-HEP-1 cells. Moreover, the effects of HIS-4 on the migration and invasion ability of HepG2 and SK-HEP-1 cells were evaluated by wound healing assay and Transwell assay. In addition, MTT assay, flow cytometry analyses, Hoechst staining, wound healing assay, Transwell assay, and tube formation assay were used to explore the anti-angiogenic activity of HIS-4 in human umbilical vein endothelial cells(HUVECs). Mechanistically, the HIS-4 regulatory of signal pathways in H9 epG2 and SK-HEP-1 cells were analyzed by Western blot. This results showed that HIS-4 suppressed the proliferation of human hepatoma HepG2 and SK-HEP-1 cells. Moreover HIS-4 induced their apoptosis of HepG2 and SK-HEP-1 cells. HIS-4 inhibited the migration and invasion of HepG2 and SK-HEP-1 cells. Additionally, HIS-4 exhibited angiogenesis effects. Mechanistically, up-regulation of MAPK signaling pathway and down-regulation of mTOR signaling pathway may be responsible for anti-hepatoma activity of HIS-4. Therefore, HIS-4 may be a promising candidate drug for HCC treatment.


Subject(s)
Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Biflavonoids , Pharmacology , Carcinoma, Hepatocellular , Drug Therapy , Pathology , Cell Movement , Cell Proliferation , Dracaena , Chemistry , Hep G2 Cells , Liver Neoplasms , Drug Therapy , Pathology , Phytochemicals , Pharmacology
9.
Article in Chinese | WPRIM | ID: wpr-777484

ABSTRACT

Stroke has become the second leading cause of death in the world, and the most common type is the ischemic stroke. Due to its rapid onset and complex conditions, ischemic stroke is a major neurological disorder that causes disability. Ischemic stroke mainly results from atherosclerosis, and the pathogenesis of ischemic stroke mainly includes energy metabolism disorders in the brain, the toxicity of excitatory amino acids, oxidative/nitrification stress, inflammatory response, apoptosis, and autophagy. With the characteristics of multi-component and multi-target, traditional Chinese medicine could be used to treat ischemic stroke at different stages. This article summarized the latest research progress on the pathogenesis of ischemic stroke and commonly used traditional Chinese medicine for treatment of ischemic stroke in order to provide references for the further research and clinical treatment of ischemic stroke.


Subject(s)
Humans , Brain Ischemia , Therapeutics , Medicine, Chinese Traditional , Research , Stroke , Therapeutics
10.
Article in Chinese | WPRIM | ID: wpr-771696

ABSTRACT

The purpose of this study was to investigate the effect of Huaier on autophagy of human hepatoma SK-HEP-1 cells and the effect of autophagy on the proliferation of SK-HEP-1 cells. CCK-8 assay was used to evaluate the effect of Huaier on the proliferation of SK-HEP-1 cells under different concentrations and different times. Acridine orange staining was used to measure the effect of Huaier on the autolysosome formation in SK-HEP-1 cells. Immunofluorescence assay was applied to examine the effect of Huaier on the expression and distribution of autophagy marker LC3 in SK-HEP-1 cells. In addition, LC3 expression was also checked by immunoblot analysis in the presence of Huaier. At last, the effects of Huaier in combination with autophagy inhibitor bafilomycin A1 on the proliferation of SK-HEP-1 cells was detected by CCK-8 assay. The results showed that Huaier aqueous extract significantly inhibited the proliferation of human hepatoma SK-HEP-1 cells in a dose- and time-dependent manner. Huaier aqueous extract dramatically promoted the formation of autolysosome in SK-HEP-1 cells. Moreover, Huaier markedly increased the number and intensity of intracellular LC3 fluorescent puncta and up-regulated LC3-Ⅱ expression. These data indicated that Huaier evidently activated autophagy of SK-HEP-1 cells. Additionally, autophagy inhibition significantly attenuated the sensitivity of SK-HEP-1 cells to Huaier treatment. Therefore, autophagy activation is involved in the inhibitory effects of Huaier on the proliferation of human hepatoma SK-HEP-1 cells.


Subject(s)
Humans , Apoptosis , Autophagy , Carcinoma, Hepatocellular , Drug Therapy , Pathology , Cell Line, Tumor , Cell Proliferation , Complex Mixtures , Pharmacology , Up-Regulation
11.
Article in Chinese | WPRIM | ID: wpr-275182

ABSTRACT

The purpose of this study was to investigate the effect and mechanism of Huaier aqueous extracton growth and invasion of human prostate cancer PC3 cells. CCK-8 assay was used to evaluate the inhibitory effect of Huaier aqueous extract on proliferation of PC3 cells. The effects of Huaier aqueous extract on cell cycle and apoptosis of PC3 cells were analyzed by flow cytometry. Moreover, wound healing assay and Transwell assay were performed to determine the effect of Huaier aqueous extract on invasion and migration abilities of PC3 cells. PC3 cells treated with Huaier aqueous extract were subjected to western blotting for protein levels of EMT markers and phosphorylation levels of key proteins in MAPK pathway. Results revealed that Huaier aqueous extract significantly inhibited the proliferation of PC3 cells in a dose-dependent and time-dependent manner. Huaier aqueous extract dramatically increased the apoptosis rate and induced S-phase arrest in PC3 cells.Furthermore, Huaier suppressed invasion and migration abilities of PC3 cells, and facilitated MET process of PC3 cells via down-regulation of N-cadherin and TCF8/ZEB1 and up-regulation of E-cadherin. In addition, Huaier reduced the phosphorylation of JNK and ERK. Therefore, the regulatory effects of Huaier on EMT and MAPK pathway may be responsible for the suppressive effect of Huaier on growth and invasion of PC3 cells.

12.
Article in Chinese | WPRIM | ID: wpr-236039

ABSTRACT

Huaier (Trametes robiniophila) has been widely used as an adjuvant drug for cancer treatment in China. The anti-cancer effect of Huaier extract has been confirmed in liver cancer, lung cancer, breast cancer, ovarian cancer, gastric cancer, and so on. The main mechanisms by which Huaier exerts an anti-neoplastic effect include inhibition of the growth and proliferation of cancer cells, induction of apoptosis of cancer cells, suppression of angiogenesis, inhibition of the invasion and migration of cancer cells, regulation of oncogenes and tumor suppressor genes expression, improving immunity, and reversal of drug resistance in cancer cells. In order to provide references for further study and clinical application on anti-tumor effect of Huaier, the latest research progress on anti-tumor effect of Huaier in recent years is summarized in this paper.


Subject(s)
Animals , Humans , Antineoplastic Agents , Pharmacology , Trametes
13.
Article in Chinese | WPRIM | ID: wpr-339617

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the correlation between the ratio of serum vascular endothelial growth factor (VEGF)/endostatin (ES) and childhood acute leukemia(AL).</p><p><b>METHODS</b>Serum levels of VEGF and ES were measured using ELISA in 35 children with acute AL before and after chemotherapy. The ratio of VEGF/ES was calculated. Thirty healthy children served as the control group.</p><p><b>RESULTS</b>The serum levels of VEGF (196 ± 66 pg/mL vs 29 ± 10 pg/mL) and ES (35 ± 7 ng/mL vs 19 ± 4 ng/mL) in the AL group before chemotherapy were significantly higher than those in the control group (P<0.01). The ratio of VEGF/ES in the AL group before chemotherapy was significantly higher than that in the control group (6.7 ± 3.0 vs 1.6 ± 0.7; P<0.01). In 20 children with AL who achieved complete remission, the serum levels of VEGF and ES and the VEGF/ES ratio were reduced after chemotherapy (83 ± 35 pg/mL, 27 ± 5 ng/mL, 3.1 ± 1.3, respectively; P<0.01), although the serum levels of VEGF and ES were still higher than those in the control group (P<0.01). The VEGF/ES ratio in CR patients was not significantly different from that in the control group. The serum levels of VEGF (r=0.301, P=0.045) and the VEGF/ES ratio (r=0.411, P=0.015) were positively correlated with the count of blast cells in juvenile bone marrow in 35 children with AL before chemotherapy.</p><p><b>CONCLUSIONS</b>Serum VEGF and ES levels are associated with the development of childhood AL. The VEGF/ES ratio can be used to evaluate the disease progression in children with AL.</p>


Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Acute Disease , Endostatins , Blood , Leukemia , Blood , Drug Therapy , Vascular Endothelial Growth Factor A , Blood
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