Effect and Mechanism of Wumeiwan Against Lewis Lung Cancer in Mice with Syndrome of Cold and Heat in Complexity Based on HGF/C-Met Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect and mechanism of Wumeiwan against Lewis lung cancer in mice with syndrome of cold and heat in complexity based on hepatocyte growth factor/mesenchymal-epithelial transition factor （HGF/C-Met） signaling pathway. MethodTwenty healthy male mice were classified into blank group， model group （equivalent volume of distilled water， ig）， cisplatin group （4.0 mg·kg-1 cisplatin， ip）， and Wumeiwan group （12.5 mL·kg-1 Wumeiwan， ig）， with 5 in each group. Lewis lung cancer with the syndrome of cold and heat in complexity was induced in mice except the blank group by gavage of propylthiouracil， Zhimu Shigaotang， and Fanxieye， ice-water swimming， and subcutaneous injection of dry yeast suspension and Lewis cell suspension under the right armpit. After modeling， administration began and lasted 6 weeks. After the experiment， the tumor weight， tumor volume， tumor inhibition rate， and lung cancer metastasis-inhibiting proportion were measured and calculated. The pathological morphology of lung tissue was observed based on hematoxylin and eosin （HE） staining. The growth state of tumor tissue was analyzed by immunohistochemistry. The mRNA expression of HGF and C-Met was detected by Real-time polymerase chain reaction （PCR）， and the protein expressions of HGF， C-Met， survivin， and X-linked inhibitor of apoptosis protein （XIAP） by Western blot. ResultCompared with the blank group， the model group showed high mRNA expression of HGF and C-Met and protein expression of HGF， C-Met， surviving， and XIAP （P<0.01）. Compared with the model group， Wumeiwan group displayed low proportion of positive cells， positive cell density， positive score （P<0.05）， histochemical score， tumor weight， tumor volume （P<0.01）， mRNA expression of HGF and C-Met （P<0.01）， and protein expression of HGF， C-Met， surviving， and XIAP （P<0.01）. Compared with the model group， the cisplatin group displayed decrease in the proportion of positive cells， density of positive cells （P<0.05）， positive score， tumor weight， tumor volume （P<0.01）， mRNA expression of HGF and C-Met （P<0.01）， and protein expression of HGF， C-Met， surviving， and XIAP （P<0.01）， and insignificant variation in the histochemical score. Wumeiwan group had high mRNA expression of HGF （P<0.01）， and insignificant variation in the proportion of positive cells， positive cell density， histochemical score， positive score， tumor weight， tumor volume， mRNA expression of C-Met， and protein expression of HGF， C-Met， surviving， and XIAP. ConclusionWumeiwan can slow down the progression of Lewis lung cancer in mice with syndrome of cold and heat I complexicity by inhibiting HGF/C-Met signaling pathway.

Mechanism of Chinese herbal formula QHF against breast cancer MCF-7 cells invasion and migration / 中草药·英文版

Objective: To investigate the effects of Chinese herbal formula Qinghuofu (QHF) on the migration and invasion of breast cancer MCF-7 cells and its possible molecular mechanisms, thereby providing a theoretical basis to find effective anti-cancer medicine and therapeutic targets for the treatment of anti-migration and anti-invasion of breast cancer. Methods: Breast cancer MCF-7 cells were treated with different QHF and other different reagents, CCK8 assay was used to detect the influence of the reagents on the proliferation of MCF-7 cells; Scrape migration and Transwell assay were used to quantitatively determine the migration and invasion effects of QHF and hepatocyte growth factor (HGF) on the MCF-7 cells. Subsequently, the c-Met inhibitor and its downstream ERK and PI3K inhibitors were used to investigate the relationship between the migration and invasion of MCF-7 cells, as well as its downstream MAPK/ERK and PI3K/Akt signaling pathways. The expression levels of HGF, c-Met, ERK, p-Akt, p-c-Met, p-ERK, p-Akt, MMP2, MMP9, and VEGF in breast cancer MCF-7 cells treated with QHF and other reagents were also examined. Results: The result indicated that formula QHF not only significantly inhibited the proliferation of MCF-7 cells, but also significantly suppressed the effects of HGF (40 ng/mL) on the proliferation and movement of MCF-7 cells, reducing the ability of the cells to invade and migrate. Western blot analysis indicated that QHF and c-Met inhibitor significantly decreased the expression of p-c-Met, p-ERK1, p-ERK2, p-Akt, MMP-2, MMP-9, and VEGF, while HGF significantly increased the expression of p-c-Met in MCF-7 cells; c-Met downstream ERK and PI3K inhibitors also significantly decreased the expression of MMP-2, MMP-9, and VEGF in MCF-7 cells; But the difference among c-Met, PI3K, ERK, and QHF group were not statistically significant. Conclusion: QHF can prevent the proliferation, migration, and invasion of MCF-7 cells by inhibiting the HGF/c-Met and its downstream PI3K/Akt and MAPK/ERK signaling pathways; Thereby down-regulating the expression of HGF, p-Met, p-ERK1, p-ERK2, p-Akt, MMP-2, MMP-9, and VEGF.

Role of HGF/c-Met signaling pathway in crizotinib-induced apoptosis of different lung carcinoma cell lines / 中国病理生理杂志

AIM: To investigate the role of HGF/c-Met signaling pathway in crizotinib-induced apoptosis of different lung carcinoma cell lines and to analyze its potential regulatory mechanisms .METHODS: EML4-ALK positive cell line H2228, c-Met proliferation cell line H1993 and control cell line A549 were treated with crizotinib at different doses for different time periods .The viability of the cell lines was measured by MTT assay .The apoptosis was analyzed by flow cytometry with PI staining.The protein levels of MET and phosphorylated MET (p-MET) of HGF/c-Met signaling pathway as well as its down-stream key proteins AKT , ERK, p-AKT and p-ERK in the cell lines before and after crizotinib treatment were examined by Western blot .RESULTS:The growth of H1993, H2228 and A549 cell lines was inhibited after crizoti-nib treatment for 72 h in a dose-dependent manner .Apoptotic rates of H1993 cells and H2228 cells were increased with the crizotinib concentration and exposure time .Down-regulation of p-MET, p-AKT and p-ERK at protein levels in H1993 cells and H2228 cells after exposure to crizotinib for 72 h was confirmed by Western blot .No obvious change of the related-pro-teins of HGF/c-Met signaling pathway was found in A 549 cell line.CONCLUSION: HGF/c-Met signaling pathway may contribute to crizotinib-induced apoptosis of H1993 cells and H2228 cells, which provides the experimental basis for MET-targeting treatment of lung cancer .

RNA Interference Targeting c-Met Inhibits Proliferation of Lens Epithelial Cells / 中国康复理论与实践

@#Objective To observe the effect of small-interfering RNA(siRNA) targeting c-Met,the receptor of hepatocyte growth factor(HGF),on the proliferation of human lens epithelial cells(LECs).Methods siRNA was transferred into LECs cultured in vitro by HiperFect Transfection Reagent.Real-Time PCR was applied to observe the expression of c-Met mRNA in LECs after gene transfer,and MTT assay was used to detect the proliferation of LECs induced by HGF.Results The expression of c-Met mRNA in LECs was significantly decreased in the experimental group,compared to that in the controls(P<0.01).Proliferation of LECs induced by HGF was inhibited,compared with the single HGF stimulated group(P<0.01).Conclusion The RNA interference targeting c-Met can effectively inhibit the expression of c-Met mRNA,and the proliferation of LECs induced by HGF.

Role of HGF/c-Met in Serum-Starved ARPE-19 Cells

PURPOSE: Hepatocyte growth factor (HGF) and its receptor (HGFR/c-Met) regulate motility, mitogenesis, and morphogenesis in a cell type-dependent fashion. We report the role of HGF and c-Met on stress-induced ARPE-19 human retinal pigment epithelial (RPE) cells in this study. METHODS: The cells were cultured either with or without serum. Southern and Western blot analyses were done to determine the expression patterns of HGF/c-Met in serum-starved ARPE-19 cells. The cell proliferation pattern in serum-starved condition was analyzed using MTS assay. Inhibition level of cell proliferation was analyzed using a neutralizing monoclonal antibody against c-Met (2 microgram/ml). RESULTS: Abnormal cell proliferation and scattering of ARPE-19 cells was observed under serum starvation. HGF/c-Met were expressed in serum-starved ARPE-19 cells. ARPE-19 cell proliferation was also enhanced with recombinant HGF treatment. Neutralization against c-Met inhibited the proliferation of serum-deprived ARPE-19 by 64.5% (n=9, S.D. 5.5%). Serum starvation appears to induce epithelial-mesenchymal transition of ARPE-19 cells, resulting in scatter, and the expression of alpha-smooth muscle actin (alpha-SMA), a marker for fibrosis. CONCLUSIONS: In conclusion, c-Met induced under non-physiologic conditions has significant effects on the activation of RPE cells.

New molecular targets and novel agents for human hepatoma / 中国药理学通报

Current anti-hepatoma agents in clinical aplication have not been proved to be satisfactory. The major obstacles are low efficacy, toxicity, and drug resistance. Identifying new drug targets and discovering new agents accordingly with high efficacies and low toxicities have become the key part of the solution. Recent studies have shown that hyper-methylation of tumor suppressor genes, interaction between hepatocyte growth factor and its receptor, vascular endothelial growth factor and its receptor, as well as cyclooxygenase-2 might be potential targets for hepatomachemotherapy. Indeed, agents acting on these targets have shown to be effective. In addition, other agents such as As 2O3 have also shown their activities against hepatoma.

New molecular targets and novel agents for human hepatoma / 中国药理学通报

Current anti-hepatoma agents in clinical aplication have not been proved to be satisfactory. The major obstacles are low efficacy, toxicity, and drug resistance. Identifying new drug targets and discovering new agents accordin gly with high efficacies and low toxicities have become the key part of the solu tion. Recent studies have shown that hyper-methylation of tumor suppressor gene s, interaction between hepatocyte growth factor and its receptor, vascular endothelial growth factor and its receptor, as well as cyclooxygenase-2 might be potential targets for hepatomachemotherapy. Indeed, agents acting on these targets have shown to be effective. In addition, other agents such as As 2O 3 have also shown th eir activities against hepatoma.