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【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.
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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.
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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.
Subject(s)
Rabbits , Autopsy , Biotechnology , Catheter Ablation , Enzyme-Linked Immunosorbent Assay , Follow-Up Studies , Hepatocyte Growth Factor , Interleukin-6 , Necrosis , Punctures , Vascular Endothelial Growth Factor AABSTRACT
<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>
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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 .
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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.
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BACKGROUND: The expression of c-Met is substantially elevated in most malignant human cancers. We therefore searched for c-Met expression and compared the expression level among malignant skin cancers. OBJECTIVE: The aim of this study was to determine the c-Met expression pattern and the protein expression level in selected malignant cutaneous tumors. METHODS: G361 cells (malignant melanoma cell line) and A431 cells (squamous cell carcinoma cell line) were cultured and analyzed, using immunoprecipitation and Western blot analysis, for expression of c-Met. Additionally, 16 separate specimens of malignant melanomas (MMs), 16 squamous cell carcinomas (SCCs), 16 basal cell carcinomas (BCCs) and 16 normal tissues were analyzed for the expression of c-Met using immunohistochemical studies. RESULTS: Based on cultured cell immunoprecipitation and Western blot analysis, both A431 cells and G361 cells expressed c-Met, however, c-Met was expressed substantially more in G361 cells. Immunohistochemical examination of c-Met showed that it was over-expressed in all malignant skin cancers. In addition, c-Met expression was more increased in MM compared to other cancers. CONCLUSION: In our study, c-Met is involved in malignant skin cancer development and the level of its expression is thought to be related to the degree of malignancy in melanoma cancers.
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Humans , Blotting, Western , Carcinoma, Basal Cell , Carcinoma, Squamous Cell , Cells, Cultured , Immunoprecipitation , Melanoma , Proto-Oncogene Proteins c-met , Skin , Skin NeoplasmsABSTRACT
@#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.
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PURPOSE: Liver transplantation is the therapy of choice for patients with acute and acute-on-chronic severe liver failure or hepatocellular carcinoma. But a suitable liver is not always available for transplantation due to limited donor numbers. To increase the number of available liver for transplantation, a non-heart-beating donor (NHBD) liver transplant program is started. In NHBD liver transplantation, warm ischemic injury of liver occurs. The duration of warm ischemia is thought to be the most important risk factor for postoperative complications such as primary nonfunction or severe hepatic dysfunction. Recent evidence indicates that hepatocyte growth factor (HGF) plays an important role as a cytoprotector against hepatic injury by anti-apoptotic effect and mitogen in liver regeneration. Therefore studies also were performed to examine whether HGF influenced the viability and regeneration of hepatocytes from rats, subjected to prolonged warm ischemic injury. METHODS: Male Sprague- Dawley rats were subjected to non-heart-beating death by cervical spine fracture. Rats left in room temperature directly after, 30-minutes, 1-hours before surgery and perfusion was performed for isolating hepatocyte. Among three groups, hepatocyte viability was compared by trypan blue stain. And isolated hepatocytes from 30-minutes warm ischemic group were cultured for 24-hours, which were treated with no HGF and addition of various doses (5 ng/mL, 10 ng/mL, 20 ng/ mL, 40 ng/mL, 100 ng/mL) of HGF. Anti-apoptosis and regeneration of hepatocyte were compared by LDH assay, MTS assay, western blot, and immunocyto-chemistry after a 24-hours culture. RESULTS: The results of hepatocyte viability along the prolonged warm ischemic groups in isolated hepatocytes decreased sequentially 74.8+/-12.6%, 45.0+/-5.4%, 37.8+/-10.4% along directly after, 30-minutes, 1-hours in trypan blue stain (P<0.01). And 24-hour-cultured hepatocytes from 30-minutes warm ischemic group were treated with HGF. The results of LDH assay, MTS assay did not have relation with HGF addition. But the results of western blot and immunocytochemistry shown that HGF doses dependent anti-apoptosis and regeneration of hepatocyte increased. That indicates HGF presumably inhibites apoptotic pathway by phosphorylation. And HGF also makes hepatocyte hypertrophy and albumin synthesis. CONCLUSION: HGF was a potent cytoprotector against hepatic injury by anti- apoptotic effect and mitogen of liver regeneration in NHBD liver animal model. HGF facilitates recovery of the liver from prolong warm ischemic injury. If the more clinical studies and large animal studies are performed, NHBD using liver transplantation will be available with more chances by HGF.
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Animals , Humans , Male , Rats , Blotting, Western , Carcinoma, Hepatocellular , Diminazene , Hepatocyte Growth Factor , Hepatocytes , Hypertrophy , Immunohistochemistry , Liver , Liver Failure , Liver Regeneration , Liver Transplantation , Models, Animal , Perfusion , Phosphorylation , Postoperative Complications , Regeneration , Risk Factors , Spine , Tissue Donors , Transplants , Trypan Blue , Warm IschemiaABSTRACT
The expression of c-met stimulated by high glucose in human renal tubular epithelial cells and the role of HGF/c-met system in diabetic nephropathy were examined. The proximal tubular epithelial cells were cultured in vitro under different conditions. MTT was used for the detection of cellular proliferation and RT-PCR was employed for measurement of c-met mRNA level. Our results showed that under different conditions, there were no significant differences in the proliferation of proximal tubular epithelial cells 12 h and 24 h after the culuture (P＞0.05). The proliferation of proximal tubular epithelial cells showed a significant change 96 h after the culture and the cellular proliferation induced by hepatocyte growth factor (HGF) was very active (P＜0.05). Moreover, no significant difference in the expression of c-met mRNA was found 12 h after the culture under different conditions (P＞0.05), while 24 and 96 h after the culture, a persistent and significantly higher expression of c-met mRNA was found in HGF-induced proliferation. It is concluded that addition of exogenous HGF could inhibit the apoptosis induced by high-level glucose, promote the proliferation of proximal tubular epithelial cells, and induce the expression of c-met. Our study suggests that local up-regulation of HGF/c-met system plays an important role in the repair of renal damage in diabetic nephropathy.
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Hepatocyte growth factor (HGF), a potent stimulator of hepatocyte growth, stimulates motility, invasiveness, proliferation, and morphogenesis of epithelium, and may be involved in physiologic and pathologic processes such as embryogenesis, wound healing, organ regeneration, inflammation, and tumor invasion. So we examined the role of the HGF/c-Met on invasion and metastasis of hypopharyngeal squamous cell carcinoma (SCC). We performed immunohistochemical stains on 40 normal mucosas and 40 SCCs of hypopharynx with HGF antibody and c-Met antibody. For RT-PCR and Western blot, fresh normal tissues and cancer tissues in hypopharynx obtained from five patients were used. The positive rates of HGF and c-Met expression in hypopharyngeal SCC were 77.5% and 70%, respectively. HGF staining was significantly correlated with lymph node metastasis and pathologic stage (p<0.05). c-Met staining was only significantly correlated with lymph node metastasis (p<0.05). The increased expresssion of c-Met mRNA (RT-PCR) and protein (Western Blot) were detected in hypopharyngeal cancer tissue. These results suggest that HGF may play an important role in the progression of hypopharyngeal cancer.
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Female , Humans , Pregnancy , Blotting, Western , Carcinoma, Squamous Cell , Coloring Agents , Embryonic Development , Epithelium , Hepatocyte Growth Factor , Hepatocytes , Hypopharyngeal Neoplasms , Hypopharynx , Inflammation , Lymph Nodes , Morphogenesis , Mucous Membrane , Neoplasm Metastasis , Pathologic Processes , Regeneration , RNA, Messenger , Wound HealingABSTRACT
Objective To explore the possible effects of hepatocyte growth factors (HGF) on prevention and therapy of disused gastrocnemius atrophy in rats by observing their effects on the wet weight and contractile function of the gastrocnemius of fixed hind limbs. Methods Twenty-four Wister male rats were randomly divided into a blank group, a contrast group and an experimental group. Their right hind limbs were fixed for four weeks with plastic piaster from metatarsus to 1 cm up the knee. Their knee joint was fixed in 100?flexion and their ankle joint in 60?plantar flexion. Rats in the contrast group were injected with 0. 2 mL of normal saline, and rats in the ex- perimental group were injected with HGF (10 mg/kg, 10 mg/mL). Their whole gastrocnemius of the right hind limb was dissected four weeks later to measure its contractile function and wet weight. Results Compared with the blank group, single maximal contraction, maximal tetanus, and wet weight of the gastrocnemius in the contrast and experimental groups decreased significantly (P＜0. 05), and fields of electric stimulation also decreased signifi- cantly (P＜0. 05). Compared with the contrast group, single maximal contraction, maximal tetanus, and wet weight of the gastrocnemius in the experimental group were significantly superior (P＜0.05), and fields of electric stimulation significantly increased. Conclusion Since after a local injection of HGF, the contractile function and wet weight can be significantly increased, HGF may have a preventive effect on disused atrophy.
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Objective:To investigate the expression and significance of hcpatocyte growth factor (HGF) and c-Met,receptor (c-Met) in endometrial carcimoma.Methods:Using S-P immunohistochemistry method,the expression of HGF and c-Met in the specimens of 52 cases with endometrial catcmoma and 16 cases with normal endometrium was examined.Then the correlation was analyzed between the two factors and the stage and differentiation of EC.Results:The expressions of HGF and c-Met in endometrial carcinoma were significantly higher than in normal endometrium.The expression of c-Met was closely associated with the stage and differentiation of endometrial carcinoma.Conclusion:Overexpression of HGF and c-Met facilitates the malignant progress of endometrium,c-Met is associated with the prognosis of EC.
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PURPOSE: Hepatocyte growth factor (HGF) is a modulator of epithelial cell proliferation and motility. In this study, we measured the level of HGF in sera and tumor extracts of gastric adenocarcinoma using an enzyme-linked immunoassay and evaluated its association with tumor progression. MATERIALS AND METHODS: The level of HGF in the sera of seventy-five patients with gastric adenocarcinoma and in the tumor extracts of forty-two tumors were examined in this study. The level of HGF was determined by an Immunis HGF EIA kit (Institute of Immunology). RESULTS: The mean level of HGF in the sera of patients was 0.26+/-0.19 ng/ml, which was significantly higher than in those of healthy controls (0.14+/-0.07 ng/ml, p<0.05); the levels of HGF in the sera of patients increased according to the progression of the stage of cancer (p<0.05). The mean level of HGF in tumor extracts was 8.22+/-9.27 microgram/g protein, which was significantly higher than in those of healthy controls (1.95+/-1.45 microgram/g protein, p<0.05); the levels of HGF in the tumor extracts were correlated significantly with the progression of the tumor stage (p<0.05). The mean level of HGF in the tumors of diffuse type was 11.28+/-11.74 microgram/g protein, which was significantly higher than in those of intestinal type (5.16+/-4.31 microgram/g protein, p<0.05). CONCLUSION: HGF may play an important role in the progression and differentiation of gastric adenocarcinoma.