The Effect Of Membrane-cytoskeleton Organizer Ezrin And CD44v6 On The Proliferation And Metastasis In Gastric Carcinoma

ObjectiveThe Ezrin protein is a member of the Ezrin,Radixin,Moesin(ERM)proteins family that are proposed to act as linkers between the cytoskeleton and plasma membrane.Ezrin is a cytoplasmic protein enriched in microvilli and other cell surface structures.lt resides on chromosome 6q25-q26.Its N-terminal domain is associated with the plasma membrane and the C-terminal domain with the cytoskeleton.Ezrin regulates cell-cell and cell-matrix interactions playing a role in the regulation of cellular adhesion,movement and morphology in epithelia.Ezrin serves as a tyrosine kinase substrate,and is phosphorylated in epidermal growth factor-stimulated cells.Ezrin is highly expressed in several types of human cancers..Recent studies have discovered that over expression of CD44v6 playd an important role in the growth,invasion,and metastasis of several kind of tumors such as colon cancer,breast cancer,and lung cancer.MethodsClinical data and tissue microarrary:Surgically resected specimens of 84 cases of gastric cancer (84 cases at the same time on the edge of greater of greater than 5cm from the pair of normal fastric mucosa as contrl),4um consecutice sections were cut,one performed conventional HE stainion,others were stored at romm temperature for further Immunohistochemistry staining.Immunohistochemical methed and assessment:S-P method was used to detect the expression of Ezrin protein and CD44v6 protein in 84 cases of gastric carcinoma,84 cases normal gastric marcinoa and 18 cases lymph node metastasis.Mouse anti-human Ezrin protein monoclonal antibody (work dilution 1:300) purchased from the company SANDA,Rabbit anti-human CD44v6 protein polylonal antibody (work dilution 1:200) purchased form Beijing bo ao shen Biologocal Technology Co.Immunostaining results assessment:Ezrin protein and CD44v6 protein reaction was defined as clearly brown granules located in the cytoplasm and membrane.For each case,200 cell were counted in high-power visual field.Staining intensity of positive cells was assessed into three graded:one point weak color;two point medium;three point height color.positivw cell≤10% zero point;positive cells 11-50% two point;positive cells 51-80% three point; positive cell>80% four point.add the two group the results were divided into four grades:if positive cells less than<10%(-);3 (+); 4-5 (++); 6-7 (+++)。Real-time quantitative results according to△CT= (experimental group-internal reference group)-(control group-internal reference group) to determine the value of the size of gastric Ezrin protein and CD44v6 protein expression.ResultsThe positive rate of Ezrin protein in gastric cancer (84.5%);were significantly higher than that in normal gastric carcinoma(61.9%) respectively,P<0.05 Ezrin protein expression in lymph node metastasis rate (88%), case and control groups were significantly different, P<0.05. In the case of the tissue typing in well-differentiated gastric cancer Ezrin protein expression rate (93.8%); poorly differentiated gastric Ezrin protein expression rate (71.4%); two type, P<0.05, there are differences. In the patient group with lymph node metastasis and without lymph node metastasis, the positive rate of,P<0.05.CD44v6 protein expression in the cell membrane, gastric cancer, the positive expression rate (75%); CD44v6 protein expression in normal gastric mucosa rate (51.2%); CD44V6 protein expression in lymph node metastasis rate (78%), case and control groups were significantly different, P<0.05. Poorly differentiated gastric cancer in the positive expression rate of CD44v6 protein (65.7%); two type P<0.05, there are differences.Quantitative RT-PCR results according to△CT= (experimental group-internal reference group)-(control group-internal reference group) to determine the value of the size of gastric Ezrin protein and CD44v6 protein expression. ConclusionEzrin protein and CD44v6 protein expression in gastric cancer group was significantly higher than in normal gastric mucosa of the control group, suggesting that Ezrin, and CD44v6 expression may be involved in the growth of gastric cancer. RT-PCR quantitative experimental results prove that in the gastric cancer group, Ezrin protein and CD44v6 protein expression of genes in a high state, that the previous results.Gastric cancer with lymph node metastasis group Ezrin protein expression was significantly higher than those without lymph node metastasis, suggesting that the expression of Ezrin protein may be involved in gastric cancer metastasis.