Chemoembolization, Therapeutic ; Humans ; Liver Neoplasms ; metabolism ; therapy ; Tetraspanin 24 ; metabolism

Antigens, CD ; genetics ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Tetraspanin 24

CD151 is a member of the tetraspanin family that is implicated as a promoter of pathological or physiological angiogenesis. C-Met is expressed on a variety of cells including vascular endothelial cells (VECs) and up-regulated during angiogenesis. In this study, we investigated whether CD151 regulated migration, proliferation, tube formation and angiogenesis of human umbilical VECs (HUVECs) with activation of C-Met. Moreover, we studied whether CD151 could affect the angiogenic molecules such as nitric oxide (NO), vascular cell adhesion molecule-1 (VCAM-1) and vascular endothelial growth factor (VEGF). The expression of CD151 was determined by Western blotting. The cell proliferation assay was performed using the cell counting kit-8 (CCK-8) method and cell migration was assessed in microchemotaxis chambers by using fetal bovine serum (FBS) as the chemotactic stimulus. The angiogenic molecules were evaluated using ELISA. The NO level was detected using NO detection kit. The potential involvement of various signaling pathways was explored using relevant antibodies. We found that proliferation, migration and tube formation of HUVECs were promoted by CD151 with activation of C-Met, FAK and CDC42, while they were suppressed with CD151 knockdown by RNAi. Similarly, the levels of NO, VCAM-1 and VEGF in HUVECs were increased by CD151, but they were inhibited with CD151 knockdown by RNAi. These data suggested that CD151 could promote migration, proliferation, tube formation and angiogenesis of HUVECs, which was possibly related to the C-Met signaling pathways.
Base Sequence ; Human Umbilical Vein Endothelial Cells ; Humans ; Neovascularization, Physiologic ; RNA, Small Interfering ; genetics ; Receptor Protein-Tyrosine Kinases ; metabolism ; Signal Transduction ; Tetraspanin 24 ; genetics ; metabolism

OBJECTIVETo investigate the expression and significance of CD151 in pituitary adenomas.

METHODSThirty-six pituitary adenomas were collected immediately after surgery together with five normal pituitary tissue. Real time-PCR, Western blot and immunohistochemistry analysis were performed to detect the expression of CD151 mRNA and protein in thirty-six pituitary adenomases and five normal pituitary tissues.

RESULTSThe expression of CD151 in all pituitary adenomases was observed to be significantly higher than that in normal pituitary tissues by Western blot, real time PCR, and immunohistochemistry analysis (P < 0.01). The expression levels of protein and mRNA in invasive pituitary adenomas were much higher than those in non-invasive pituitary adenomas (P < 0.01).

CONCLUSIONThe results suggested that the expression of CD151 was closely correlated with malignant degree of pituitary adenomas, which indicated the expression of CD151 was intimately correlated with occurrence and development of pituitary adenomas. Detecting CD151 might be a vital index to predict prognosis of pituitary adenomas.

Adenoma ; metabolism ; Blotting, Western ; Humans ; Immunohistochemistry ; Pituitary Gland ; pathology ; Pituitary Neoplasms ; metabolism ; Prognosis ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Tetraspanin 24 ; metabolism

Over-expression of CD151 was found to be associated with metastasis and poor prognosis of prostatic carcinoma. This study was designed to examine the mechanism by which CD151 promotes the proliferation and migration of prostatic cancer cells. The pAAV-CD151, pAAV-GFP and pAAV-CD151-AAA mutant plasmids were constructed and used to transiently transfect PC3 cells (a prostatic carcinoma 3 cell line) by the mediation of Fugene HD. Then, the cells were assigned to control group, pAAV-GFP group, pAAV-CD151 group, and pAAV-CD151-AAA group respectively. Cell proliferation was evaluated by using the 3-[4,5-dimet-hylthiazol-2-yl]-2,5, diphenyltetrazolium bromide (MTT) method. Cell migration assay was performed by using Boyden chambers. The formation of CD151-integrin α3/α6 complex was determined by the method of co-immunoprecipitation. The protein expression levels of CD151 and extracellular signal-regulated kinase (ERK) were measured by Western blotting. The results showed that transfection of pAAV-CD151 or pAAV-CD151-AAA mutant increased the expression of CD151 protein in PC3 cells. Co-immunoprecipitation showed that more CD151-integrin α3/α6 complex was formed in the pAAV-CD151 group than in the control group, the pAAV-GFP group and the pAAV-CD151-AAA mutant group. Furthermore, the proliferative and migrating capacity of PC3 cells was substantially increased in the pAAV-CD151 group but inhibited in the pAAV-CD151-AAA mutant group. CD151 transfection increased the expression of phospho-ERK. Taken together, it was concluded that CD151 promotes the proliferation and migration of PC3 cells through the formation of CD151-integrin complex and the activation of phosphorylated ERK.
Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Humans ; Integrin alpha3 ; metabolism ; Integrin alpha6 ; metabolism ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; Tetraspanin 24 ; metabolism

OBJECTIVETo investigate the effects of in vivo CD151 gene transfer on angiogenesis and heart function in rats with myocardial infarction.

METHODSAcute myocardial infarction (AMI) was induced in male Sprague-Dawley (SD) rats by left anterior descending coronary artery ligation. The surviving rats randomly received myocardial injection of saline (MI control), pAAV-CD151 and pAAV-GFP (n = 12/group). Sham-operated rats without myocardial injection (n = 12) were taken as normal control. Four weeks later, heart function and the expression of CD151 were measured. Micro vessels density (MVD) in infarct myocardium was observed by factor VIII related antigen immunochemical staining.

RESULTSThe expression of CD151 (1.98 +/- 0.23 vs. 0.91 +/- 0.09, P < 0.01) and MVD counting [(385.4 +/- 79.9) vs. (252.5 +/- 43.0) n/mm(2), P < 0.01] in pAAV-CD151 treated MI rats were significantly higher than that in MI control group, similarly, EF (64.0 +/- 8.7)% vs. (41.5 +/- 5.0)%, P < 0.01] and dp/dt(max) (6620.2 +/- 884.6 vs. 5545.5 +/- 693.0, P < 0.01) were also significantly increased post pAAV-CD151 treatment. These parameters were not affected by pAAV-GFP treatment.

CONCLUSIONCD151 in vivo gene transfer for rats with acute myocardial infarction enhanced myocardial angiogenesis and improved left ventricular function.

Animals ; Antigens, CD ; genetics ; Disease Models, Animal ; Genetic Therapy ; Genetic Vectors ; Male ; Myocardial Infarction ; physiopathology ; therapy ; Neovascularization, Physiologic ; Rats ; Rats, Sprague-Dawley ; Tetraspanin 24 ; Transfection ; Ventricular Function, Left

This study was designed to determine the effects of the recombinant adeno-associated virus vector containing sense CD151 gene (rAAV-CD151) and antisense CD151 gene (rAAV-antiCD151) on the migration of Tca8113 cell. Functional fragment of CD151 gene was amplified by RT-PCR, and inserted into the vector pAAV in the sense direction and antisense direction, respectively. The rAAV-CD151 and rAAV-antiCD151 were produced and the titers were determined by dot blot. The CD151, at protein level, was detected by Western blot. The Transwell chamber was used to detect the effects of the rAAV-CD151 and rAAV-antiCD151 on the tumor cell migration. The titers of the rAAV-CD151 and rAAV-antiCD151 were 2 x 10(11) pfu/ml and 1.0 x 10(11) pfu/ml, respectively. The expression of CD151 was increased by 108% in the cells transfected with rAAV-CD151 and decreased by 79% in the cells transfected with rAAV-antiCD151, as compared with non-transfected cells, respectively. The number of the migrating cells was significantly increased in the cells transfected with rAAV-CD151 (93.56 +/- 11.59) and decreased in the cells transfected with rAAV-antiCD151 (24.00 +/- 4.36) as compared with non-transfected and rAAV-GFP transfected cells (53.00 +/- 6.56 and 46.00 +/- 7.00, P<0.05). It is an important molecular mechanism of the tumor metastasis that the overexpression of CD151 promotes the migration of the tumor cells. The rAAV-antiCD151 is a novel tool, which can reduce the expression of CD151 and inhibit the migration of the tumor cells, and brings us a new approach of anti-sene gene therapy targeted at CD151 in human carcinoma.
Antigens, CD ; immunology ; pharmacology ; Carcinoma, Squamous Cell ; pathology ; Cell Movement ; DNA, Antisense ; pharmacology ; Dependovirus ; genetics ; Genetic Vectors ; Humans ; Recombinant Proteins ; biosynthesis ; genetics ; Tetraspanin 24 ; Tongue Neoplasms ; pathology ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo investigate the efficacy of CD151 gene delivery in promoting blood perfusion in swines after myocardial infarction.

METHODSSwines received coronary artery ligation and intramyocardial injection with rAAV-CD151, rAAV-anti-CD151 or rAAV-GFP. Eight weeks after vector injection, Western blot, immunostaining and 13N-labeled NH3 PET were performed to detect gene expression and biological effects of various treatments.

RESULTSHigh level of CD151 protein expression was detected in the rAAV-CD151 group. The capillary density in the rAAV-CD151 group [(83.8 +/- 6.7) n/mm2] was significantly higher than that in the control group [(33.2 +/- 4.5) n/mm2] and rAAV-GFP group [(41.6 +/- 5.6) n/mm2] (all P<0.05); the arteriole density in the rAAV-CD151 group [(16.4 +/- 2.5) n/mm2] was also higher than that in the control group [(6.6 +/- 2.3) n/mm2] and the rAAV-GFP group [(8.4 +/- 1.6) n/mm2] (all P<0.05). However, the lowest capillary density and arteriole density were evidenced in rAAV-anti-CD151 group. Myocardial blood perfusion was significantly increased in rAAV-CD151 group and significantly reduced in rAAV-anti-CD151 group (all P<0.05 vs. control).

CONCLUSIONIntramyocardial injection of rAAV-CD151 could enhance the myocardial express of CD151 protein, increase capillary and arteriole densities and improve blood perfusion in swine with myocardial infarction.

Animals ; Antigens, CD ; genetics ; Coronary Occlusion ; therapy ; Dependovirus ; genetics ; Female ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; Humans ; Male ; Myocardial Infarction ; therapy ; Neovascularization, Physiologic ; Swine ; Swine, Miniature ; Tetraspanin 24 ; Treatment Outcome