The effect of ALPL on liver regeneration after partial hepatectomy in mice / 西安交通大学学报(医学版)

【Objective】 To observe the role of liver/bone/kidney alkaline phosphatase gene (ALPL) in liver regeneration following 70% hepatectomy (partial hepatectomy, PH). 【Methods】 A knock-out mouse model (ALPL+/-) was established, and a 70% hepatectomy was performed. Changes in liver weight and liver function were measured at PH 1 day, PH 3 day, and PH 7 day (PH1d、PH3d、PH7d) after surgery. In addition, cell proliferation, hepatocyte growth factor (HGF), and vascular endothelial growth factor (VEGF) were performed by Western blotting, immunofluorescence staining, and enzyme linked immunosorbent assay. 【Results】 ALPL knockout mice at PH7d exhibited a lower ratio of liver/total body weight than normal control mice. An analysis of liver function showed no significant difference between the ALPL knockout group and the WT (ALPL+/+) group when the ALPL gene was deleted. While Ki67 staining and PCNA analysis indicated that liver cell proliferation was decreased in ALPL+/- mice at PH1d and increased at PH7d compared to that in ALPL+/+group. Additionally, knockouts of ALPL decreased serum and liver HGF and VEGF levels at PH1d compared to WT controls, but increased at PH7d. 【Conclusion】 The knockout of ALPL leads to a delayed liver regeneration following hepatectomy, which provides theoretical support for exploring the mechanisms underlying liver regeneration after hepatectomy.

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.

MET overexpression and intratumor heterogeneity in esophageal squamous cell carcinoma

Esophageal squamous cell carcinoma (ESCC) is among the ten most frequent and deadly cancers, without effective therapies for most patients. More recently, drugs targeting deregulated growth factor signaling receptors have been developed, such as HGF-MET targeted therapy. We assessed MET and HGF genetic alterations and gene and protein expression profiles in ESCC patients from the Brazilian National Cancer Institute and publicly available datasets, as well as the intratumor heterogeneity of the alterations found. Our analyses showed that HGF and MET genetic alterations, both copy number and mutations, are not common in ESCC, affecting 5 and 6% of the cases, respectively. HGF showed a variable mRNA expression profile between datasets, with no alterations (GSE20347), downregulation (GSE45670), and upregulation in ESCC (our dataset and GSE75241). On the other hand, MET was found consistently upregulated in ESCC compared to non-tumor surrounding tissue, with median fold-changes of 5.96 (GSE20347), 3.83 (GSE45670), 6.02 (GSE75241), and 5.0 (our dataset). Among our patients, 84% of the tumors showed at least a two-fold increase in MET expression. This observation was corroborated by protein levels, with 55% of cases exhibiting positivity in 100% of the tumor cells. Intratumor heterogeneity was evaluated in at least four tumor biopsies from five patients and two cases showed a consistent increase in MET expression (at least two-fold) in all tumor samples. Our data suggested that HGF-MET signaling pathway was likely to be overactivated in ESCC, representing a potential therapeutic target, but eligibility for this therapy should consider intratumor heterogeneity.

Assessment of impact of varied preconditioning factors, used in isolation and combination, for enhancement of cytokine secretions in stem cell conditioned medium

Stem cell conditioned medium exhibits a huge regenerative potential but low concentration of cytokines, highdevelopment cost, and scalability challenges are major deterrents to product advances. This research studiedimpact on vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) secretions afterpreconditioning human dental pulp stem cells (HDPSCs) with diverse factors, viz., Deferoxamine (250 µM),Epidermal Growth Factor (EGF) (2 ng/ml), Angiotensin I (5mM) and Insulin Transferrin Selenium (ITS) (1%),and Niacinamide (5 mM). Additionally, advantage of using “pooled population” of HDPSCs was ascertained.HDPSCs were incubated under standard culture conditions using optimized growth media. On reaching 80%–85% confluency, media was discarded and fresh serum free media with preconditioning factors, initially oneat a time and subsequently in combination, was added. Spent media, collected after 48 hours of incubation,was used for enzyme-linked immunosorbent assay testing of selected cytokines. In comparison to control,preconditioning factors, used in isolation showed varied but significant upregulation in the secretion of VEGFbut down regulation of HGF. When used in combination, VEGF secretion increased almost 10 times of controlwith no significant change in HGF level. Results demonstrated importance of choosing right preconditioningfactors and right interplay of preconditioning factors to enhance secretions. This research paves the path todevelop an effective and commercially viable conditioned medium that can be used as “biological active” forpharmaceuticals and for developing “customized” serum free medium enriched with selected cytokines

Research progress on the mechanism of resistance to MET inhibitors in non-small cell lung cancer and review of MET inhibitor resistance-related strategies / 中国肿瘤临床

The MET gene is an important tumor-driving gene for non-small cell lung cancer (NSCLC). Drugs targeting tumor with MET exon 14 skipping mutations bring new hope for patients. Although MET inhibitors such as tepotinib and savolitinib have shown good antitumor effects, resistance is inevitable. Studies on the hepatocyte growth factor (HGF)/mesenchymal- epithelial transition factor (MET) signaling pathway will not only help explore the mechanism underlying resistance to MET inhibitors, they may aid in the discovery of strategies for inhibiting and reversing drug resistance, thereby expanding the field of novel drug development. Preliminary studies have shown that the combination of HGF/MET inhibitors with other drugs may have great potential for clinical applications. This article reviews the characteristics of MET gene abnormalities, the mechanism of resistance against MET inhibitors, and the strategies for responding to resistance. Finally, the challenges posed by MET inhibitors is discussed and guidance on the direction of future development of MET inhibitors is proposed.

Yacon Extract Attenuated Kidney Fibrosis in 5/6-subtotal Nephrectomy Mouse Model by Upregulating HGF and BMP-7 mRNA Expression

@#Introduction: Chronic kidney disease (CKD) leads to tubular injury, kidney fibrosis and anemia. These conditions are influenced by fibrotic and anti-fibrotic substances, such as Transforming Growth Factor beta-1 (TGF-β1), Hepatic Growth Factor (HGF), and Bone Morphogenic Protein-7 (BMP-7). Yacon is an herbal medicine which has not been elucidated in CKD. This study aimed to investigate the effect of ethanolic extract of Yacon leaves on attenuating renal injury in CKD model in mice. Methods: We performed 5/6 subtotal nephrectomy (SN) in male Swiss-Webster mice (3 months old, 30–40 grams) to induce chronic kidney disease, then the mice were sacrificed at day 14. The mice (n=25) were divided into five groups: one SN group, three groups of SN with administration of Yacon extract, and one group of sham operation (SO, with supplementation of 0.1% aquadest). There were three different doses of ethanolic extract of Yacon leaves: 98 mg/kg BW (SN+YK1), 49 mg/kg BW (SN+YK2), and 24.5 BW mg/kg (SN+YK3). Tubular injury, perivascular and interstitial fibrosis were quantified based on histopathological examination. Reverse-transcriptase PCR (RT-PCR) was performed to quantify HGF and BMP-7. Results: SN group demonstrated CKD with elevation of creatinine level, anemia, tubular injury, glomerulosclerosis, and fibrosis. Yacon extract treatment showed attenuation of injury with lower creatinine level, tubular injury, glomerulosclerosis and fibrosis compared to the SN group. HGF and BMP-7 mRNA expressions were higher in Yacon-treated groups than the SN group. Conclusion: Yacon treatment might ameliorate CKD through reducing fibrosis and increasing expression of anti-fibrotic genes.

miR-130a-3p inhibits invasion of breast cancer MCF-7 cells through HGF/MET pathway / 中国肿瘤生物治疗杂志

@# Objective: To explore the molecular mechanism of miR-130a-3p regulating epithelial mesenchymal transition (EMT) to affect the invasion and metastasis of breast cancer cells through HGF/MET pathway. Methods: A total of 22 pairs of cancer tissues and adjacent normal tissues from breast cancer patients, who were admitted to Affiliated Hospital of Chengde Medical College from January 2018 to October 2018, were collected for this study; in addition, breast cancer cell lines (MCF-7, MDA-MB-231 and MDA-MB453) and normal breast epithelial cells MCF10A were obtained from the Institute of Basic Sciences, Chengde Medical College. And then, the expression of miR-130a-3p in tissues and cell lines were detected by qRT-PCR. The experiment cells were divided into control group, miR-130a-3p mimics group, miR-130a-3p inhibitor group, PHA665752 (a small-molecule MET inhibitor) transfection group and PHA665752+miR-130a-3p inhibitor co-transfection group. CCK-8 assay and Transwell assay were performed to detect the proliferation, invasion and migration of MCF-7 cells, respectively. The expressions of EMT and HGF/MET signaling pathway related proteins in MCF-7 cells were detected by WB. In addition, the targeted relationship between miR-130a-3p and MET was verified by Dual luciferase reporter gene assay. Results: miR-130a-3p was down-regulated in breast cancer tissues and cell lines. Over-expression of miR130a-3p could suppress the proliferation, invasion, migration and EMT of MCF-7 cells, while knockdown of miR-130a-3p had the opposite results. The results of Dual luciferase reporter gene assay indicated that miR-130a-3p targetedly down-regulated the expression of MET, and miR-130a-3p negatively regulated the expression of HGF/MET signaling pathway. Further experiments confirmed that miR-130a-3p inhibited the proliferation, invasion, migration and EMT of MCF-7 cells by blocking HGF/MET signaling pathway. Conclusion: miR-130a-3p suppresses the EMT of MCF-7 cells via blocking HGF/MET signaling pathway, thereby repressing the invasion and metastasis of MCF-7 cells.

Hepatocyte growth factor and carotid intima-media thickness in relation to circulating CD34-positive cell levels

BACKGROUND@#Hepatocyte growth factor (HGF) may act as a possible biochemical index for vascular damage, although evidence for the association between HGF and carotid intima-media thickness (CIMT) is limited. Since both HGF and circulating CD34-positive cells play an important role in endothelial repair, circulating CD34-positive cell levels may influence the association between HGF and CIMT.@*METHODS@#We conducted a cross-sectional study of 269 elderly Japanese men aged 60-69 years who had undertaken an annual medical checkup from 2014 to 2015.@*RESULTS@#The median value for circulating CD34-positive cells was 0.93 cells/μL. Among the study population, 135 men showed low circulating CD34-positive cell levels (≤ 0.93 cells/μL). By multivariable linear regression analysis, HGF was found to be significantly positively associated with CIMT only to participants with low circulating CD34-positive cell levels, with a multi-adjusted β of 0.26 (p = 0.005) and 0.002 (0.986) for low and high circulating CD34-positive cell levels, respectively. In addition, a significant interaction was observed between HGF and circulating CD34-positive cell levels (low and high) on CIMT (multivariable p value of 0.049). A positive association exists between HGF and CIMT in elderly Japanese men, limited to participants with low circulating CD34-positive cell levels.@*CONCLUSION@#A positive association exists between HGF and CIMT in community-dwelling elderly Japanese men, which is limited to participants with low numbers of circulating CD34-positive cells. Our findings indicate that circulating CD34-positive cell levels could determine the influence of HGF on CIMT in elderly Japanese men.

Protective effects of total saponins of Panax japonicas on non-alcoholic fatty liver disease through regulating expression of miR-199-5p / 中国中药杂志

To research the effection and probable mechanism for the total saponins of Panax japonicas(TPSJ) in mice on non-alcoholic fatty liver disease. Forty SPF male Kunming mice were randomily divided into four group:control group,NAFLD group, low-dose TPSJ treated group,high-dose TPSJ treated group. High-fatty and high-frutose-diet was applied to eatablish NAFLD model，and TPSJ (100 and 200 mg·kg⁻¹) in feeding were given for the TPSJ groups for 4 weeks. To collect the serum with liver and the ALT and TC of serum were monitored after 4 weeks. The hepatic histopathologic structure was observed by haematoxylin-eosin (HE) staining, RT-PCR and RT-qPCR was applied for the detection of miR-199-5p，VEGFa，HGF，c-Met and protein expression level was detected bv laser confocal microscope.Compared with control group, the level of serum ALT and TC in the model group was higher,the liver of the model group showed that hepatocytes display obvious lipid deposition. Then TPSJ treated showed that markedly improved histopathologic changes, decreased fatty deposition. In the meantime,the expression level of miR-199-5p was significantly decreased, thus the expression of HGF and c-Met were significantly increased. TPSJ play a role of prevention on fatty liver, the machanism maybe by blocking miR-199-5p targeted to c-Met signaling pathways in NAFLD.

No-Touch Radiofrequency Ablation of VX2 Hepatic Tumors In Vivo in Rabbits: A Proof of Concept Study

OBJECTIVE: In a proof of concept study, we compared no-touch radiofrequency ablation (NtRFA) in bipolar mode with conventional direct tumor puncture (DTP) in terms of local tumor control (LTC), peritoneal seeding, and tumorigenic factors, in the rabbit VX2 subcapsular hepatic tumor model. MATERIALS AND METHODS: Sixty-two rabbits with VX2 subcapsular hepatic tumors were divided into three groups according to the procedure: DTP-RFA (n = 25); NtRFA (n = 25); and control (n = 12). Each of the three groups was subdivided into two sets for pathologic analysis (n = 24) or computed tomography (CT) follow-up for 6 weeks after RFA (n = 38). Ultrasonography-guided DTP-RFA and NtRFA were performed nine days after tumor implantation. LTC was defined by either achievement of complete tumor necrosis on histopathology or absence of local tumor progression on follow-up CT and autopsy. Development of peritoneal seeding was also compared among the groups. Serum hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) were measured via ELISA (Elabscience Biotechnology Co.) after RFA for tumorigenic factor evaluation. RESULTS: Regarding LTC, there was a trend in NtRFA (80%, 20/25) toward better ablation than in DTP-RFA (56%, 14/25) (p = 0.069). Complete tumor necrosis was achieved in 54.5% of DTP-RFA (6/11) and 90.9% of NtRFA (10/11). Peritoneal seeding was significantly more common in DTP-RFA (71.4%, 10/14) than in NtRFA (21.4%, 3/14) (p = 0.021) or control (0%). Elevations of HGF, VEGF or IL-6 were not detected in any group. CONCLUSION: No-touch radiofrequency ablation led to lower rates of peritoneal seeding and showed a tendency toward better LTC than DTP-RFA.

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.

Effects of acupuncture at opposite acupoints on expression of hepatocyte growth factor in rats with skeletal muscle contusion / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of acupuncture at opposite acupoints on expression of hepatocyte growth factor (HGF) in rats with skeletal muscle contusion, and to explore the mechanism of acupuncture at opposite acupoints on skeletal muscle contusion.</p><p><b>METHODS</b>Fifty-four Sprague Dawley (SD) rats were randomly divided into a blank group (6 rats), a model group (24 rats) and an opposing needling group (24 rats). The model group and opposing needling group were further divided into 1-day subgroup, 3-day subgroup, 5-day subgroup and 7-day subgroup, 6 rats in each one. No intervention was given in the blank group, while the model of skeletal muscle contusion was established in the model group and opposing needling group by self-made contusion device. 24 hours after contusion, electroacupuncture (EA) was applied at "Zusanli" (ST 36) and the corresponding points ofpoints at health side for 15 min, once a day. The subgroups of opposing needling group were treated for 1 day, 3 days, 5 days and 7 days, respectively. No treatment was given in the model group. Samples were collected in the subgroups 1 day, 3 days, 5 days and 7 days after treatment. The morphological change of injured gastrocnemius muscle was observed by using microscope after HE staining. The positive cell rate of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry. The expression levels of HGF protein and PCNA protein were observed by Western blot.</p><p><b>RESULTS</b>① The results of HE staining showed that, 1 day after contusion, the inflammatory cells of gastrocnemius muscle in the opposing needling group were less than those in the model group; 3 days and 5 days after contusion, myoblasts and myotubes in the opposing needling group were more than those in the model group; 7 days after contusion, the neonatal muscle cells in the opposing needling group were more than those in the model group. ② The results of immunohistochemistry showed that, 1 day, 3 days and 5 days after contusion, the positive cell rate of PCNA in the opposing needling group was significantly higher than that in the model group (all<0.001); 7 days after contusion, the positive cell rate of PCNA in the opposing needling group was significantly less than that in the model group (<0.001). ③ The results of Western blot showed that, 1 day, 3 days and 5 days after contusion, the expression of HGF protein and PCNA protein in the opposing needling group was significantly higher than that in the model group (all<0.05); 7 days after contusion, the expression of HGF protein and PCNA protein in the opposing needling group was significantly lower than that in the model group (all<0.05).</p><p><b>CONCLUSION</b>Acupuncture at opposite acupoints could regulate the expression of HGF and promote the activation, proliferation, migration and differentiation of muscle satellite cells in rats with skeletal muscle contusion, which could speed up the process of skeletal muscle injury repair.</p>

Role of Met/HGF signal pathway on inhibiting autophagy in ischemia-reperfusion induced cardiomyocytes injury / 安徽医科大学学报

Objective To investigate the effect and mechanism of autophagy inhibited by Met/HGF passway in alleviating myocardial ischemia and reperfusion injury.Methods Lsolated cardiomyocytes of rats were divided into 5 groups:control group,ischemia and reperfusion (IR) group,Met-transfection group,Met siRNA-transfection group,Met-transfection and HGF activator group.Over or less expressions of Met were controlled by the method of transfection.Cell proteins were extracted after IR injury.The expression of cell protein such as met,HGF,AMPK,PI3K,autophagy-related protein LC3B,Beclin-1 and apoptosis-related protein Bcl-2 were detected by Western blot.Results Compared with control group,Western blot results showed that the expression of AMPK and PI3K increased significantly by the method of over expression of Met and activation of HGF,the expression of autophagy-related protein LC3B and Beclin-1 decreased significantly at the same time.However,it showed an opposite trend in IR group and Met siRNA-transfection group.Conclusion Activation of the Met/HGF signal pathway inhibits autophagy and attenuates myocardial ischemia-reperfusion injury.

Application of iTRAQ multiplex tandem mass spectrometry in the expression of HGF in breast cancer cell invasion / 国际检验医学杂志

Objective To investigate the clinical value of iTRAQ multiplex tandem mass spectrometry in the detection of the ex-pression of hepatocyte growth factor (HGF) in patients with invasive breast cancer .Methods A total of 35 patients with breast cancer and 30 healthy subjects were selected from January 2014 to October 2016 in this hospital ,the expression of serum HGF in breast cancer patients with different clinical stages and healthy subjects was analyzed by iTRAQ labeling ,mass spectrometry ,library searching and Scqffold software ,and the differential expression of HGF was verified by Western blot .Results A total of 237 pro-teins were identified in the serum samples of this study ,and 89 proteins with strict quantitative criteria ,17 differentially expressed proteins ,included HGF ,were screened for breast cancer patients and healthy controls .iTRAQ markers showed that the expression level of serum HGF in different clinical stage of breast cancer patients was significantly higher than that in healthy subjects (P<0 .05) .The results of Western blot showed that the relative expression level of serum HGF in breast cancer patients was significant-ly higher than that in healthy subjects(P<0 .05) .Conclusion iTRAQ multiplex tandem mass spectrometry is useful for the detection of breast cancer patients with high expression of HGF ,which is of great significance in guiding the clinical treatment of breast cancer .

Application of iTRAQ multiplex tandem mass spectrometry in the expression of HGF in breast cancer cell invasion / 国际检验医学杂志

Objective To investigate the clinical value of iTRAQ multiplex tandem mass spectrometry in the detection of the ex-pression of hepatocyte growth factor (HGF) in patients with invasive breast cancer .Methods A total of 35 patients with breast cancer and 30 healthy subjects were selected from January 2014 to October 2016 in this hospital ,the expression of serum HGF in breast cancer patients with different clinical stages and healthy subjects was analyzed by iTRAQ labeling ,mass spectrometry ,library searching and Scqffold software ,and the differential expression of HGF was verified by Western blot .Results A total of 237 pro-teins were identified in the serum samples of this study ,and 89 proteins with strict quantitative criteria ,17 differentially expressed proteins ,included HGF ,were screened for breast cancer patients and healthy controls .iTRAQ markers showed that the expression level of serum HGF in different clinical stage of breast cancer patients was significantly higher than that in healthy subjects (P<0 .05) .The results of Western blot showed that the relative expression level of serum HGF in breast cancer patients was significant-ly higher than that in healthy subjects(P<0 .05) .Conclusion iTRAQ multiplex tandem mass spectrometry is useful for the detection of breast cancer patients with high expression of HGF ,which is of great significance in guiding the clinical treatment of breast cancer .

Interstitialcells of liver cancer influence the biological function of malignant through the HGF/cMET system / 西安交通大学学报(医学版)

Objective To explore the effects of interstitial cells of liver cancer and normal liver cells co‐cultured on the biological function of liver cancer malignancy so as to understand the signal pathway involved by the interaction between these cells and confirm the role of interstitial cells in cancer progression in tumor microenvironment .Methods We co‐cultured interstitial cells or hepatocyte growth factor (HGF ) and human normal liver cell L‐02 ,and then detected the expressions of the tumor‐suppressing gene PTEN and the oncogene K‐RAS and changes of cell proliferation .The downstream signaling pathways were detected by Real‐time PCR and Western blot .Results The expression of PTEN was downregulated at the transcription level by 1 .15 times and translation level by 10 times (P<0 .05) ,while the transcription level and translation level of K‐RAS increased by 1 .4 times and more than 9 times , respectively ( P< 0 .05 ) in normal liver cells co‐cultured with liver cancer mesenchymal cells .The proliferation ability was increased by more than 2 times .ELISA experiment results showed that the medium from co‐culture contained HGF over 3 times more than the control group ( P<0 .05 ,1 085+108 vs .387+23) .At the same time ,cells in the experimental group expressed more than four times of cMET than the control group cells (P< 0 .05) .Exogenous HGF consistently promoted liver cell proliferation and viability (P<0 .05) .Conclusion Our study shows that liver cancer interstitial cells activate the HGF/cMET signaling pathway by secreting HGF and promote the proliferation of normal liver cells ,suggesting a new way to explore the molecular mechanism of tumor microenvironment in tumor development and treatment of hepatocellular carcinoma .

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 .

Evaluation of therapeutic effect of Ad-HGF gene modified PMSCs on limb ischemia in rabbit model / 军事医学

Objective To evaluate the therapeutic effect of hepatocyte growth factor(HGF) gene modified placenta-derived mesenchymal stem cells( PMSCs) on limb ischemia in a rabbit model.Methods The placental tissue was digested with enzyme, cultured and passaged.The PMSCs were characterized by surface marker expression.These cells were infected with adenoviral( Ad)-HGF and intramuscular injected for treatment of limb ischemia in a rabbit model.The blood supply of the limb was detected by digital subtraction angiography ( DSA ) and the vessel number was evaluated in histopathological HE staining.Results The results showed that Ad-HGF gene transduction increased the vascular endothelial growth factor ( VEGF ) , basic fibroblast growth factor, bFGF ( bFGF ) and HGF expression in PMSCs. Transplantation of HGF-transduced PMSCs resulted in the increase in vessel density and improvement of blood supply in the rabbit limb ischemia model.Conclusion The therapeutic effect of HGF gene engineered PMSCs on ischemia by enhancing angiogenesis in a rabbit model is evaluated.Transplantation of PMSCs with HGF gene therapy may be a promising strategy for the treatment of ischemia diseases.

Expressions of C-Met and HGF in tongue squamous cell carcinoma and their clinical significance / 西安交通大学学报(医学版)

ABSTRACT:Objective To study the expressions of C-Met and hepatocyte growth factor (HGF)in tongue squamous cell carcinoma and their clinical significance.Methods We analyzed 46 patients (the treatment group) with tongue squamous carcinoma and 27 patients (the control group)with benign tumor treated in our hospital between June 201 1 and May 2013.The expressions of C-Met and HGF were detected by immunohistochemistry. Results In the experimental group,C-Met negative expression rate was 21.7% (10/46),weak positive expression rate was 26.1% (12/46),and strong positive expression rate was 52.2% (24/46).The above expression rates were 77.8% (21/27),1 5.8% (5/27),and 3.7% (1/27)in the control group.The two groups differed significantly (P <0.001).In the experiment group,the negative expression of HGF protein was 1 7.4% (8/46 ),weak positive expression rate was 58.7% (27/46),and strong positive expression rate was 23.9% (1 1/46).The above expression rates in the control group was 63.0% (1 7/27 ),37.0% (10/27 ),and 0.0% (0/27 ).The two groups differed significantly (P <0.001).The expression of C-Met protein was significantly related to lymph node metastasis (P <0.05).The expression of HGF protein was significantly related to pathological grading and lymph node metastasis (P <0.05).Conclusion The expressions of C-Met and HGF in tongue squamous cell carcinoma are significantly higher.And the occurrence and development of tongue squamous cell carcinoma are closely related to the expressions of C-Met and HGF.

Correlation research between prognosis of NSCLC patient and serum levels of HGF and IL-6 / 局解手术学杂志

Objective To study the correlation between prognosis of non-small cell lung cancer ( NSCLC) patient and serum levels of HGF and IL-6. Methods Eligible 109 NSCLC patients and preoperative blood samples of each patient in our hospital from March 2013 to May 2014 were collected. Then the levels of HGF and IL-6 of serum samples were detected by using ELISA kit. All patients were followed up for 1 year. The neoplasm staging:67 cases were stagingⅠ-Ⅱ,38 cases were stagingⅢ,2 cases were staging IV. There were 79 cases with ad-enomatous carcinoma( ADC) ,26 cases with squamous carcinoma( SCC) and 4 cases with NSCLC. According to the TNM staging,there were 79 cases in pN( -) group,31 in pN( +) group,77 in pT(T1-T2) group and 28 in pT(T3-T4) group. Results The average concentration of HGF and IL-6 in serum were 860 pg/mL and 2. 7 pg/mL respectively. Analysis of survival indicated that,compared to those patient with lower serum level of HGF and IL-6,the survival rate of patient with high serum level was much lower. The difference was statistically signifi-cant (HGF,P=0. 019;IL-6,P=0. 002). The analysis of the patients with stageⅢdisease separately also indicated that survival rate of pa-tient with lower serum of HGF and IL-6 was higher than others. Conclusion The serum levels of HGF and IL-6 might be a effective indica-tors for predicting prognosis of the NSCLC paitents.