Humans ; Proto-Oncogene Proteins c-met ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism

This research aims to construct a lentiviral expression vector carrying the extracelluar domain (ED) of human hepatocyte growth factor receptor (C-Met), and to express it in transfected 293T cells. The extracellular domain of C-Met was amplified by RT-PCR, ligated with lentiviral expression vector p RRL-CMV-ED, and then expressed in 293T cell line. The expressed protein was purified and identified by RT-PCR and Western blot. The enzyme digestion and sequence analysis showed that the lentiviral expression vector p RRL-CMV-ED was constructed correctly. The size of amplified genes was about 2 700 bp. The purified protein with Ni-affinity column was about 105 kD analyzed by SDS-PAGE. The Western blot and ELISA results showed that the expressed protein which could bind to HGF specifically was the extracelluar domain of human hepatocyte growth factor receptor. This research may lay a foundation for further study of anti-C-MET monoclonal antibody and neutralizing antibody.
Genetic Vectors ; HEK293 Cells ; Humans ; Lentivirus ; Proto-Oncogene Proteins c-met ; genetics ; metabolism ; Transfection

Recently, targeted therapy has achieved great success in the treatment of non-small cell lung cancer (NSCLC) patients. Mesenchymal to epithelial transition factor (MET) is considered to be another important molecular target for NSCLC since epidermal growth factor receptor (EGFR) and anaplastic lymphoma kinase (ALK). Accumulating clinical trials and case reports have confirmed that MET inhibitors exhibited a potential prospect in treating patients with MET 14 exon skipping alterations, suggesting that MET 14 exon skipping mutation might be an effective biomarker for MET inhibitors, which remains to be confirmed by more clinical data. This review summarizes current research about the molecular mechanism, clinicopathological characterization, treatment strategies and drug resistance mechanisms of MET 14 exon skipping alterations in NSCLC.
.
Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; Exons ; genetics ; Humans ; Lung Neoplasms ; drug therapy ; genetics ; Molecular Targeted Therapy ; Mutation ; Proto-Oncogene Proteins c-met ; genetics

The mesenchymal-epithelial transition factor (MET) exon 14 skipping mutation is mainly caused by the loss of c-Cbl tyrosine binding site. This mutation could result in a decrease in the degradation rate of proteasome-mediated MET proteins, trigger continuous activation of downstream pathways, and ultimately lead to tumorigenesis. The incidence of MET exon 14 skipping mutation in patients with non-small cell lung cancer (NSCLC) is 0.9% to 4.0%. Patients with advanced NSCLC are recommended to test MET exon 14 skipping mutations who may benefit from MET inhibitors-targeted therapy. MET inhibitors have a high objective response rate and good safety profiles, which could prolong the survival of NSCLC patients with MET exon 14 skipping mutations. The Lung Cancer Specialty Committee of Chinese Elderly Health Care Association organized multidisciplinary experts to give suggestions on the important issues of clinical aspects for targeted therapy of MET exon 14 skipping mutation in NSCLC according to the clinical practice experiences and evidences based medicine. "Expert Consensus on Targeted Therapy of NSCLC with MET Exon 14 Skipping Mutation" is proposed, aiming to provide standardized guidances for the clinical practice of Chinese physicians.
.
Humans ; Aged ; Carcinoma, Non-Small-Cell Lung/genetics* ; Lung Neoplasms/genetics* ; Consensus ; Proto-Oncogene Proteins c-met/genetics* ; Mutation ; Exons ; Protein Kinase Inhibitors/therapeutic use*

Humans ; Carcinoma, Non-Small-Cell Lung/genetics* ; China ; Consensus ; East Asian People ; Immunohistochemistry ; Lung Neoplasms/genetics* ; Proto-Oncogene Proteins c-met/genetics*

Mesenchymal-epithelial transition factor (MET) amplification is an important driver of resistance in epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer (NSCLC), and the combination of MET proto-oncogene (MET) and EGFR-tyrosine kinase inhibitors (TKIs) has shown promise in overcoming this molecularly defined acquired resistance. Emerging data also demonstrate MET amplification as a resistance driver to TKIs-treated anaplastic lymphoma kinase (ALK)-, RET-, and ROS1-fusion NSCLC. Here, we review the literature on recent research progress of MET amplification as a resistance driver to targeted therapy in oncogene-driven NSCLC and summarize the progress of clinical strategies to overcome the resistance mechanism.
.
Carcinoma, Non-Small-Cell Lung/genetics* ; Drug Resistance, Neoplasm/genetics* ; ErbB Receptors/genetics* ; Humans ; Lung Neoplasms/pathology* ; Mutation ; Protein Kinase Inhibitors/therapeutic use* ; Protein-Tyrosine Kinases ; Proto-Oncogene Proteins/genetics* ; Proto-Oncogene Proteins c-met/genetics*

OBJECTIVETo explore the effect of c-met ribozyme transfection on biological behavior of gastric carcinoma cells.

METHODSU1/met292 plasmid containing c-met ribozyme gene was transfected into L2 subline of SGC-7901 gastric cell line, and the proliferative ability, distribution of cell cycle, protein expression of VEGF and c-met, as well as the potential of liver metastasis of the transfected subline were determined.

RESULTSThere were no significant difference in proliferative ability, distribution of cell cycle between the transfected cells and the control cells. The protein expression of VEGF and c-met, as well as the liver metastatic potential significantly decreased in the transfected cells than those in the control cells (P< 0.05).

CONCLUSIONThe liver metastatic potential of c-met positive gastric cancer cells may be prevented by inhibiting c-met expression.

Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Plasmids ; Proto-Oncogene Proteins c-met ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transfection ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo explore the effects of c-met-siRNA on the growth and invasion of hepatocellular carcinoma MHCC97-H cells by pSuppressorRetro/c-met-siRNA recombinant plasmid transfection.

METHODSRecombinant plasmid transfection to Phoenix A cells was constructed using the lipofectin method and then the retrovirals containing c-met-siRNA were used to infect target cells MHCC97-H. In vitro, c-met expression was tested by Western blot. Cell proliferation, motility and invasiveness were studied using MTT, cell migration assay, and cell invasion assay, respectively.

RESULTSThe expression of c-met decreased significantly in MHCC97-H cells, and the most effective site of the target sequence was at 537. The growth, motility and invasiveness of MHCC97-H cells were inhibited.

CONCLUSIONThe results indicate that c-met-siRNA can down-regulate the expression of c-met and inhibit hepatocellular carcinoma cell proliferation, motility and invasiveness.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Plasmids ; Proto-Oncogene Proteins c-met ; genetics ; metabolism ; RNA, Small Interfering

Targeted therapy is an important therapeutic method for advanced non-small cell lung cancer with driver gene alteration.However,resistance to targeted therapy will inevitably happen in clinical practice,which has become a major issue demanding prompt solution.Studies have demonstrated that bypass resistance mediated by the activation of hepatocyte growth factor(HGF)/mesenchymal-epithelial transition factor(MET)signaling pathway is a common cause of resistance to targeted therapy.Presently,relevant studies have accumulated rich experience in the specific mechanisms.To be brief,HGF/MET is an important target for overcoming the resistance to targeted therapy and promises to be a leading biomarker for judging and observing the occurrence of resistance.This paper introduces the recent studies concerning the effects and mechanisms of HGF/MET signaling pathway on resistance to targeted therapy.
Carcinoma, Non-Small-Cell Lung/genetics* ; Epithelial-Mesenchymal Transition ; Hepatocyte Growth Factor ; Humans ; Lung Neoplasms/genetics* ; Proto-Oncogene Proteins c-met/metabolism* ; Signal Transduction

OBJECTIVE: To explore the effects of c-Met-siRNA on the proliferation, movement and invasion of laryngeal carcinoma Hep-2 cells in vitro. METHOD: Firstly, the pSilencer 2.0/c-Met-shRNA recombinant plasmid was transfected into laryngeal carcinoma Hep-2 cells with transfecting agent of cationic liposome Lipofectamine 2000. Secondly,the transfection efficacy was tested by RT-PCR and Western-Blot, then the most inhibitive c-Met-siRNA sequence was elected. Cell proliferation, movement and invasion were detected with MTT, cell migration assay and cell invasion assay, respectively. RESULT: After the transfection of pSilencer 2.0/c-Met-shRNA recombinant plasmid into laryngeal carcinoma Hep-2 cells, the expression of mRNA and protein of c-Met decreased significantly in Hep-2 cells, and ability of the proliferation, movement and invasion of laryngeal carcinoma Hep-2 cells were also inhibited. CONCLUSION The results indicated that c-Met-siRNA can down-regulated the expression of c-Met and markedly inhibited laryngeal carcinoma Hep-2 cell proliferation, movement and invasion. It may have the potential as a therapeutic modality to treat human laryngeal carcinoma.
Apoptosis ; genetics ; Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; Liposomes ; Proto-Oncogene Proteins c-met ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection