Anastrozole promotes implantation by altering the expression of paxillin and fak in rat luminal uterine epithelium / El anastrozol promueve la implantación alterando la expresión de paxilina y fak en el epitelio uterino luminal de rata

An alternative hyper-ovulator inducer to replace clomiphene citrate (CC) is needed as it is unsuitable for women with polycystic ovarian syndrome and is associated with low pregnancy rates. Anastrozole is an effective hyper-ovulator inducer, but has not been well researched. In order to determine the effectiveness of anastrozole as a hyper-ovulator inducer and to an extent compare it with CC in similar situations, this study ascertained the effects of these drugs on the expression of the focal adhesion proteins, paxillin and FAK, which are uterine receptivity markers in the surface luminal uterine epithelial cells of day 1 and day 6 pregnant Wistar rats. The results show that paxillin is localized in focal adhesions at the base of the uterine epithelial cells at day 1 of pregnancy whereas at day 6, paxillin disassembles from the basal focal adhesions and localizes and increases its expression apically. FAK is faintly expressed at the basal aspect of the uterine epithelial cells while moderately expressed at the cell-to-cell contact at day 1 in all groups from where it disassembles and relocates apically and becomes more intensely expressed at day 6 of pregnancy in untreated and anastrozole treated rats. Although paxillin is localized apically at day 6, its expression is significantly down-regulated with CC treatment suggesting its interference with the implantation process. These findings seem to suggest that anastrozole could favor implantation.

Para reemplazar el citrato de clomifeno (CC) es necesario un inductor de hiperovulación alternativo, ya que no es adecuado para mujeres con síndrome de ovario poliquístico y está asociado con tasas bajas de embarazo. El anastrozol es un inductor eficaz del hiper-ovulador, pero no se ha investigado adecuadamente. Con el fin de determinar la efectividad del anastrozol como inductor del hiper-ovulador y, en cierta medida, compararlo con CC en situaciones similares, este estudio determinó los efectos de estos fármacos en la expresión de las proteínas de adhesión focal, paxillin y FAK, uterinas marcadores de receptividad en la superficie luminal de células uterinas epiteliales, del día 1 y día 6 en ratas Wistar preñadas. Los resultados muestran que la paxilina se localiza en adherencias focales en la base de las células epiteliales uterinas en el día 1 del embarazo, mientras que en el día 6, la paxilina se desmonta de las adherencias focales basales y localiza y aumenta su expresión apicalmente. FAK se expresa débilmente en el aspecto basal de las células epiteliales uterinas, mientras que se expresa moderadamente en el contacto de célula a célula en el día 1 en todos los grupos, donde se separa y se reubica apicalmente y se expresa con mayor intensidad el día 6 de la preñez, en pacientes no tratados y tratados. ratas tratadas con anastrozol. Aunque la paxillina se localiza apicalmente en el día 6, su expresión está significativamente disminuida con el tratamiento con CC, lo que sugiere su interferencia con el proceso de implantación. Estos hallazgos sugieren que el anastrozol podría favorecer el proceso de implantación.

Effect of downregulation of phosphatase and tensin homolog gene expression on p130crk- related substrate protein and paxillin signal transduction in activated hepatic stellate cells in vitro / 中华肝脏病杂志

Objective@#To investigate the role of adenovirus-mediated short hairpin RNA (shRNA) in down-regulating the expression of phosphatase and tensin homologue deleted on chromosome ten (PTEN) on p130Crk-related substrates(p130Cas) and paxillin signal transduction to activate hepatic stellate cell (HSC) in vitro.@*Methods@#The rat hepatic stellate cell line, HSC-T6 was cultured and activated in vitro. The adenovirus was used as a vector to transiently transfect shRNA targeting PTEN to activate HSC in vitro, and then PTEN low expression model of activated HSC in vitro was established. Western blot and real-time fluorescence quantitative PCR were used to detect the protein and mRNA expression of PTEN, p130cas and paxillin in activated HSC. The experiment was divided into control group (HSC were transfected with DMEM medium instead of adenovirus), Ad-GFP group (HSC were infected with empty the adenovirus expressing green fluorescent protein (GFP) alone), and Ad-shRNA/PTEN group (HSC were infected with the recombinant adenovirus containing both shRNA targeting PTEN and GFP gene). One-way analysis of variance was used for comparison of multiple groups, and LSD test was used for inter-group comparison.@*Results@#shRNA targeting PTEN was successfully transfected and significantly down-regulated the PTEN protein and mRNA expression of HSC in vitro (P < 0.05), and the PTEN low expression model of HSC in vitro was successfully constructed. Compared with the expression of p130cas mRNA in the three groups of HSC, the expression fold of p130cas mRNA in the Ad-GFP group and the Ad-shRNA / PTEN group was 1.01 times and 1.52 times, respectively. The expression of p130cas mRNA in HSC of the Ad-shRNA / PTEN group was significantly higher than control group and Ad-GFP group (P < 0.05), but there was no statistically significant difference between the control group and the Ad-GFP group (P > 0.05). The expression of p130cas protein in the three groups was higher than that in the control group (0.74 ± 0.07) and the Ad-GFP group (0.72 ± 0.02); P < 0.05, but there was no statistically significant difference between the Ad-GFP group and the control group (P > 0.05). The expression of paxillin mRNA in the three groups of HSCs was compared with the expression of paxillin mRNA in the control group of HSC being 1, the expression folds of paxillin mRNA in the Ad-GFP group and Ad-shRNA / PTEN group were 0.97 times and 1.58 times, respectively. The expression of paxillin mRNA in the Ad-shRNA / PTEN group was higher than that in the control group and the Ad-GFP group (P < 0.05), and there was no statistically significant difference between the control group and the Ad-GFP group (P > 0.05). The expression of paxillin protein in the three groups of HSCs was higher in the Ad-shRNA / PTEN group (0.91 ± 0.05) than control group (0.46 ± 0.03) and Ad-GFP group (0.50 ± 0.04), P < 0.05, and there was no statistically significant difference between the Ad-GFP group and the control group (P > 0.05).@*Conclusion@#Down-regulation of PTEN expression can significantly boost p130cas and signal transduction activity of paxillin protein in activated HSC in vitro.

Trypanosoma cruzi down-regulates mechanosensitive proteins in cardiomyocytes

BACKGROUND Cardiac physiology depends on coupling and electrical and mechanical coordination through the intercalated disc. Focal adhesions offer mechanical support and signal transduction events during heart contraction-relaxation processes. Talin links integrins to the actin cytoskeleton and serves as a scaffold for the recruitment of other proteins, such as paxillin in focal adhesion formation and regulation. Chagasic cardiomyopathy is caused by infection by Trypanosoma cruzi and is a debilitating condition comprising extensive fibrosis, inflammation, cardiac hypertrophy and electrical alterations that culminate in heart failure. OBJECTIVES Since mechanotransduction coordinates heart function, we evaluated the underlying mechanism implicated in the mechanical changes, focusing especially in mechanosensitive proteins and related signalling pathways during infection of cardiac cells by T. cruzi. METHODS We investigated the effect of T. cruzi infection on the expression and distribution of talin/paxillin and associated proteins in mouse cardiomyocytes in vitro by western blotting, immunofluorescence and quantitative real-time polymerase chain reaction (qRT-PCR). FINDINGS Talin and paxillin spatial distribution in T. cruzi-infected cardiomyocytes in vitro were altered associated with a downregulation of these proteins and mRNAs levels at 72 h post-infection (hpi). Additionally, we observed an increase in the activation of the focal adhesion kinase (FAK) concomitant with increase in β-1-integrin at 24 hpi. Finally, we detected a decrease in the activation of FAK at 72 hpi in T. cruzi-infected cultures. MAIN CONCLUSION The results suggest that these changes may contribute to the mechanotransduction disturbance evidenced in chagasic cardiomyopathy.

Regulation of paxillin tyrosine phosphorylation via inhibition of c-Abl kinase to protect ventilator induce lung injury in vivo in rats / 中华危重病急救医学

Objective To determine whether the inhibition of paxillin tyrosine residues 31 and tyrosine residues 118 (Pxn Y31 and Pxn Y118) phosphorylation via inhibition of c-Abl kinase will effectively block its downstream effector molecules vessel endothelium-cadherin (VE-cad), and whether Rho/Rho kinase activation which will induce the vascular barrier dysfunction. Methods Ninety healthy male Sprague-Dawley (SD) rats were randomly divided into nine groups (each n =10). Only tracheotomy was undergone in the sham group. Groups of protective ventilation were set at a volume tidal (VT) of 6 mL/kg, a positive end-expiratory pressure (PEEP) of 5 cmH2O (1 cmH2O =0.098 kPa) for 1 hour or 2 hours (namely group PVT 1 h and group PVT 2 h), respectively. Groups of high VT were put on mechanical ventilation (MV) at high VT 30 mL/kg, PEEP 0 for 1 hour or 2 hours (namely group HVT 1 h and group HVT 2 h), respectively. Groups UO126 and AG957 pretreatment were set on MV at HVT for 1 hour or 2 hour respectively, but they were given p42/44 mitogen-activated protein kinase (p42/44MAPK) inhibitor UO1261 mg/kg by intraperitoneal injection or c-Abl kinase inhibitor AG95710 mL/kg by intragastric injection 1 hour before HVT ventilation. All the animals were sacrificed after experiments and specimens of lung tissues and bronchoalveolar lavage fluid (BALF) were harvested. Pulmonary vascular permeability was measured by Evans blue (EB). The levels of tumor necrosis factor-α(TNF-α) in BALF were measured by enzyme linked immunosorbent assay (ELISA). Then the change of lung tissue pathology was observed with light microscope, diffuse alveolar damage system (DAD) score and lung wet/dry ratio (W/D) were estimated. The myeloperoxidase (MPO) activity was measured by colorimetric analysis, phosphorylations of c-Abl Y245, Pxn Y31, Pxn Y118, VE-cad Y658, p42/44MAPK Y202/Y204, myosin light chain (MLC) and myosin-associated phosphatasetype Y696 (MYPT Y696) were determined by Western Blot. Results ① There were no obvious pathological changes in the lung tissue in the sham group and PVT 1 h or 2 h group, and also there were no significant differences in all the parameters between above groups. However, the injury in lung tissue was severe in the HVT groups. In addition, DAD score, lung W/D ratio, EB content, the activity of MPO, and TNF-α in BALF in HVT groups were significantly higher than those in sham group and PVT groups. After pretreatment with AG957 or UO126, all the parameters were significantly decreased as compared with those of groups HVT. ② The levels of phosphorylation of the proteins in lung tissue in HVT groups were increased as compared with those of group sham and groups PVT, especially at 2 hours of MV. However, compared with groups HVT, the level of p-VE-cad Y658 in lung tissue decreased significantly in group AG957 and group UO126 at 2 hours after HVT. However, the levels of all phosphorylated proteins at 2 hours were significantly lowered in the AG957 group compared with those of the HVT group [p-c-Abl Y245 (gray value): 0.29±0.04 vs. 0.42±0.04, p-Pxn Y31 (gray value): 0.51±0.03 vs. 0.70±0.05, p-Pxn Y118 (gray value):0.65±0.04 vs. 0.91±0.04, p-VE-cad Y658 (gray value): 0.77±0.07 vs. 1.32±0.07, p-p42/44MAPK Y202/Y204 (gray value): 0.38±0.06 vs. 0.61±0.03, p-MLC (gray value): 0.37±0.04 vs. 0.77±0.05, p-MYPT Y696 (gray value):0.54±0.05 vs. 0.87±0.06, all P < 0.05]. After pretreatment with UO126, the phosphorylation level of VE-cad in lung tissue at 2 hours was significantly lower than that of HVT group (gray value: 0.74±0.04 vs. 1.32±0.07), and the phosphorylation levels of p42/44MAPK and its downstream effector molecules MLC and MYPT Y696 were also significantly decreased [p-p42/44MAPK Y202/Y204 (gray value): 0.38±0.07 vs. 0.61±0.03, p-MLC (gray value):0.37±0.04 vs. 0.77±0.05, p-MYPT Y696 (gray value): 0.55±0.05 vs. 0.87±0.06, all P < 0.05]. Conclusions Pxn Y31 and Pxn Y118 phosphorylation could be blocked by inhibition of c-Abl kinase, which could strengthen VE-cad at attachment junction and might block formation of Pxn-guanine nucleotide-exchange factor H1 (GEF-H1)-p44/42MAPK signalosome which induce activation local Rho signaling, lead to activation of MLC phosphorylation, actomyosin contraction, and increase endothelial permeability.

Comparative Effects of Non-Thermal Atmospheric Pressure Plasma on Migration and Invasion in Oral Squamous Cell Cancer, by Gas Type

PURPOSE: The fourth state of matter, plasma is known as an ionized gas with electrons, radicals and ions. The use of non-thermal plasma (NTP) in cancer research became possible because of the progresses in plasma medicine. Previous studies on the potential NTP-mediated cancer therapy have mainly concentrated on cancer cell apoptosis. In the present study, we compared the inhibitory effect of NTP on cell migration and invasion in the oral squamous cancer cell lines. MATERIALS AND METHODS: We used oral squamous cancer cell lines (SCC1483, MSKQLL1) and different gases (N₂, He, and Ar). To investigate the mechanism of plasma treatment, using different gases (N₂, He, and Ar) which induces anti-migration and anti-invasion properties, we performed wound healing assay, invasion assay and gelatin zymography. RESULTS: The results showed that NTP inhibits cancer cell migration and invasion of oral squamous cancer cell. In addition, focal adhesion kinase expression and matrix metalloproteinase-2/9 activity were also inhibited. CONCLUSION: The suppression of cancer cell invasion by NTP varied depending on the type of gas. Comparison of the three gases revealed that N₂ NTP inhibited cell migration and invasion most potently via decreased expression of focal adhesion kinase and matrix metalloproteinase activity.

Estudo da participação das proteínas Paxilina e Miosina-va na infectividade do Vírus Linfotrópico de Células T Humanas do tipo 1 (HTLV-1) / Study participation of Paxillin proteins and myosin-va in infectivity of the virus lymphotropic Human T cells type 1 (HTLV-1)

As ORFs I e IV do genoma do HTLV-1 codificam, respectivamente, as proteínas p12/p8 (acessória) e Tax (regulatória). p12/p8, de 99 aminoácidos, pode ser clivada em sua extremidade amino terminal gerando a proteína p8. A primeira clivagem proteolítica de p12 remove o sinal de retenção ao RE, enquanto a segunda clivagem, gera o produto de 8kDa, referido como p8. p12 localiza-se no sistema de endomembranes, residindo em RE e aparato de Golgi, enquanto p8 dirige-se para a membrana plasmática, onde é recrutada para a sinapse imunológica, através da ligação com o receptor de células T (TCR), além de participar da sinapse virológica e da formação de conduítes. A proteína Tax, por outro lado, atua como transativador transcricional do HTLV-1, sendo referida também na indução da expressão de diversos genes celulares, aumentando a proliferação e a migração das células infectadas. Na via de transporte de vesículas secretórias, vesículas produzidas como pós-Golgi são transportadas ao longo do citoesqueleto por motores celulares. A Miosina-Va, um motor não convencional, transporta diversos cargos, incluindo vesículas secretórias, vesículas sinápticas e de retículo endoplasmático. Outra proteína relacionada ao citoesqueleto é a Paxilina, que atua como molécula adaptadora nas adesões focais e cuja expressão está aumentada em indivíduos TSP-HAM...

HTLV-1 ORFs I and IV encode respectively p12/p8 (accessory protein) and Tax (regulatory protein). The 99 amino acid p12 protein can be proteolytically cleaved at the amino terminus to generate the p8 protein. The first proteolytic cleavage removes the ER retention/retrieval signal at the amino terminus of p12, while the second cleavage generates the p8 protein. The p12 protein localizes to cellular endomembranes, within the ER and Golgi apparatus, while p8 traffics to lipid rafts at the cell surface and is recruited to the immunological synapse upon T-cell receptor (TCR) ligation, virological synapse and conduits. Tax on the other hand acts as viral transactivator and induces expression of many cellular genes, increasing proliferation and migration of infected cells. In secretory vesicle transport, vesicles produced as post-Golgi are moved along the cytoskeleton by motor proteins. The unconventional myosin motor, Myosin-Va, moves several cargoes including secretory vesicles, synaptic vesicles, and the endoplasmic reticulum. Another cytoskeleton associated protein is Paxillin, an adapter on focal adhesions which expression is increased in TSP-HAM patients...

Expression of claudin, paxillin and FRA-1 in non-nodular breast lesions in association with microcalcifications / Expressão de claudina, paxilina e FRA-1 em lesões mamárias não nodulares associadas a microcalcificações

CONTEXT AND OBJECTIVE The possible role of adhesion molecules in early breast carcinogenesis has been shown in the literature. We aimed to analyze early adhesion imbalances in non-nodular breast lesions and their association with precursor lesions, in order to ascertain whether these alterations exist and contribute towards early carcinogenesis. DESIGN AND SETTING Retrospective cross-sectional study based on medical records at a private radiological clinic in São Paulo, Brazil. METHODS We retrospectively reviewed the medical records of all consecutive women attended between August 2006 and July 2007 who presented mammographic evidence of breast microcalcifications classified as Breast Imaging Reporting and Data System Atlas (BI-RADS) type 4. These women underwent stereotaxic biopsy. Clinical, radiological and pathological data were collected, and immunohistochemical assays searched for claudin, paxillin, FRA-1 and HER-2. RESULTS Over this period, 127 patients were evaluated. Previous BI-RADS diagnoses showed that 69 cases were in category 4A, 47 in 4B and 11 in 4C. Morphological assessment showed benign entities in 86.5%. Most of the benign lesions showed preserved claudin expression, associated with paxillin (P < 0.001). Paxillin and HER-2 expressions were correlated. FRA-1 expression was also strongly associated with HER-2 expression (P < 0.001). CONCLUSIONS Although already present in smaller amounts, imbalance of adhesion molecules is not necessarily prevalent in non-nodular breast lesions. Since FRA-1 expression reached statistically significant correlations with radiological and morphological diagnoses and HER-2 status, it may have a predictive role in this setting. .

CONTEXTO E OBJETIVO A literatura tem mostrado a importância de moléculas de adesão na carcinogênese precoce de mama. Objetivamos analisar desequilíbrios precoces de adesão em lesões não nodulares da mama e associação com lesões precursoras, a fim de verificar se essas alterações existem e contribuem com a carcinogênese. TIPO DE ESTUDO E LOCAL Estudo retrospectivo baseado em prontuários médicos, numa clínica radiológica privada em São Paulo, Brasil. MÉTODOS Revisamos retrospectivamente prontuários de todas as mulheres consecutivamente atendidas com evidência mamográfica de microcalcificações mamárias, classificadas como tipo 4 do Breast Imaging Reporting and Data System Atlas (BI-RADS) entre agosto de 2006 e julho de 2007. Elas foram submetidas a biópsia estereotáxica. Dados clínicos, radiológicos e histopatológicos foram coletados e ensaios de imunoistoquímica procuraram por claudina, paxilina, HER-2 e FRA-1. RESULTADOS No período, 127 pacientes foram avaliadas. Diagnósticos de BI-RADS anteriores tinham 69 casos na categoria 4A, 47 em 4B, e 11 em 4C. A avaliação morfológica mostrou entidades benignas em 86,5%. A maioria das lesões benignas mostrou expressão preservada de claudina, associada a paxilina (P < 0,001). Expressões de paxilina e HER-2 foram correlacionadas. Expressão de FRA-1 associou-se à de HER-2 (P < 0,001). CONCLUSÕES Embora já presente em menor quantidade, o desequilíbrio de moléculas de adesão não é necessariamente prevalente em lesões mamárias nodulares e talvez a expressão de FRA-1 possa ter um papel preditivo neste cenário, uma vez que atingiu correlações ...

Expressiou and clinical significance of p130Cas and paxillin in breast carcinoma / 中国医师进修杂志

ObjectiveTo explore the expression of p130Cas and paxillin in breast carcinoma and investigate the relationships of p130Cas and paxillin levels with clinical and pathological characteristics.MethodsReverse transcriptase polymerase chain reaction (RT-PCR) was applied to detect the expression of p130Cas and paxillin in tumor tissues from 30 cases of primary breast carcinoma,15 cases of normal breast tissues and 10 cases of breast fibroadenoma.ResultsThe levels of p130Cas and paxillin were 0.444 ± 0.088,0.493 ± 0.073,0.739 ± 0.092,0.755 ± 0.137 in normal breast tissues and breast fibroadenoma tissues.The levels of p130Cas and paxillin were 0.914 ±0.186,0.303 ±0.043 in breast carcinoma tissues.Breast carcinoma tissues showed higher levels of p130Cas and lower levels of paxillin than normal breast tissues and breast fibroadenoma tissues(P ＜ 0.01 ).The expression of p130Cas was related with age (r =0.599,P =0.000 ),menopausal status (t =5.602,P =0.000 ),estrogen receptor ( ER ),progesterone receptor ( PR ) status (t =5.768,P =0.000; t =4.151,P =0.000 ) and histological grades (r =-0.668,P =0.000 ),but not related with tumor size (F =0.717,P =0.497 ),lymph node status (F =1.230,P =0.308 ) and pathological stages(r =0.283,P =0.137).The expression of paxillin was related with rumor size (F=4.114,P =0.028 ),pathological stages( r =-0.520,P =0.003),and lymph node status(F=12.418,P=0.000 ),but not related with age(r =0.294,P=0.115),menopausal status(t =-0.403,P =0.690),ER,PR status(t =0.749,P =0.460;t =0.006,P =0.995) and histological grades (r =-0.173,P =0.362).Conclusions p130Cas and paxillin have relations with the malignant transformation,invasion and metastasis of breast carcinoma.Measurement of p130Cas and paxillin may provide useful prognostic information for patients with primary or metastatic breast carcinoma.

The expression and clinical significance of Paxillin and CA Ⅸ in the benign and malignant lesions of gallbladder / 中华消化杂志

Objective To study the expression and clinicopathological significance of Paxillin and CAⅨ in the benign and malignant lesions of gallbladder.Methods The surgical resected specimens of 108 cases of gallbladder adenocarcinoma, 46 cases of peritumoral tissue, 15 cases of adenomatous polyp and 35 cases of chronic cholecystitis were routinely made paraffin embedded sections.The expressions of Paxillin and CAⅨ were stained with Envision immunohistochemistry.Results The positive rates of Paxillin and CAⅨ expression was significantly higher in gallbladder adenocarcinoma (60.2% and 49.1%) than those in peritumoral tissues (26.1%, x2 =15.00, P ＜0.01 and 23.9%,x2=8.41,P ＜0.01), adenomatous polyp (20.0%,x2=8.60,P＜0.01 and 20.0%,x2 =4.49,P＜0.05) and chronic cholecystitis(14.3% ,x2 =22.89, P＜0.01 and 11.4%,x2 =15.63,P ＜0.01).All the gallbladder epithelia of the benign cases with Paxillin and CA Ⅸ positive expression showed moderate to severe atypical hyperplasia.The positive expression rates of Paxillin and CA Ⅸ were significanctly lower in the cases of well-differentiated, maximal diameter of mass less than 2 cm, no lymph nodes metastasis and no surrounding tissues invasion than those of the cases with poorly differentiated, maximal diameter of mass over 2 cm, lymph nodes metastasis and surrounding tissues invasion.With Kaplan-Meier analysis, it suggested that the survival period after the surgery in Paxillin and CAⅨ positive expression cases was shorter than that of negative expression cases (x2 = 5.65,P＜0.05,5.65=5.92, P＜0.01).With multivariate Cox regression analysis, it indicated that both Paxillin and CAⅨ positive expression was an important indicator of the poor prognosis in gallbladder adenocarcinoma.Conclusion The expression of Paxillin and CA Ⅸ may be closely related to the carcinogenesis, tumor biological behaviors, and prognosis of gallbladder adenocarcinoma.The positive expression of Paxillin and/or CAⅨ is associated with poor prognosis.

Effects of Porphyromonas gingivalis on focal adhesive components paxillin and FAK of gingival epithelial cells / 实用口腔医学杂志

Objectives: To explore the destructive mechanism of P.gingivalis on periodontium and to better understand the pathogenic effects of P.gingivalis fimbriae. Methods:Western blotting was used to detect the degradation effects of P.gingivalis ATCC 33277 and its fimA-deficient mutant on focal adhesion components paxillin and focal adhesion kinase (FAK) of Hela cells and immortalized gingival epithelial cells (IHGE cells). Results:P.gingivalis ATCC 33277 wild strain and its fimA-deficient mutant induced degradation of paxillin and FAK both in Hela cells and in IHGE cells. fimA-deficient mutant had a remarkable weaker degradation ability than the wild strain. In IHGE cells, paxillin and FAK were degraded in a time-and MOI-dependent manner. Conclusion:P.gingivalis fimbriae-mediated adhesion and invasion to epithelial cells may promote the degradation of focal adhesion components. IHGE cells may be more suitable for the study of periodontal pathogens than Hela cells.

Expression and significance of Paxillin and VCAM-1 proteins in esophageal carcinoma / 中国综合临床

Objective To study the expression of Paxillin and VCAM-1 in esophageal carcinoma and the relationship between the expression of Paxillin, VCAM-1 and carcinogenesis and progression of esophageal carcinoma. Methods Paxillln and VCAM-1 expression were detected in 24 normal esophageal mucosa and 94 primary tumor tissues with SP immunohistochemistal method. Results The expression rate of PaxiUin was related to invasive depth (P <0.01) ,clinical staging (P <0.01) and metastasis of lymph node (P <0.05). The expression rate of VCAM-1 was related to invasive depth (P < 0.01) ,clinical staging (P < 0.01) and metastasis of lymph node (P<0.05). There was a positive correlation between Paxillin and VCAM-1 expression in this study (r = 0. 247 ,P < 0.05). Conlusion Paxillin and VCAM-1 are over expressed in esophageal carcinoma. They can be used as valuable biomakers to evaluate biological characteristics in esophageal carcinoma.

Expression of HER-2/neu and Paxillin in Ductal Carcinoma in situ, Invasive Ductal Carcinoma with Ductal Carcinoma in situ and Mucinous Carcinoma / 한국유방암학회지

PURPOSE: We purpose to determine the correlation of HER-2/neu and paxillin expression in ductal carcinoma in situ (DCIS), invasive ductal carcinoma with ductal carcinoma in situ (IDC with DCIS) and mucinous carcinoma. METHODS: To evaluate the expression of HER-2/neu and paxillin, the immunohistochemical staining was performed for 13 cases of DCIS, 13 cases of IDC with DCIS and 6 cases of mucinous carcinoma. RESULTS: The DCIS and IDC were associated with infiltration of the inflammatory cells, especially in the comedo type and solid type of tumor. In cases with infiltration of the inflammatory cells, HER-2/neu and paxillin were strongly expressed. When comparing the expression level of HER-2/neu from adjacent normal tissue between DCIS and IDC with DCIS, expression of HER-2/neu was similar to that of normal tissue adjacent to DCIS. However, in the adjoining normal ductal epithelial cells, paxillin was highly expressed in cells of all of the tumor types, and especially for IDC with DCIS. HER-2/neu and paxillin were not expressed in mucinous carcinoma cells in all cases. CONCLUSION: HER-2/neu in the DCIS and IDC with infiltration of inflammatory cells shows higher expression than non-inflammatory DCIS and IDC. If normal duct epithelial cells show a high level of HER-2/neu expression, the epithelial cells have a high probability of transformation into anaplastic cells. However, paxillin appears to have no value as a prognostic factor. The difference of expression of HER-2/neu between IDC with DCIS and DCIS suggests a different origin of tumor cells. The growth pattern of mucinous carcinoma cell is different from the that of DCIS or IDC cell, which grow slowly.

The role of the alpha4 integrin-paxillin interaction in regulating leukocyte trafficking

The movement of leukocytes from the blood into peripheral tissues is a central feature of immune surveillance, but also contributes to the pathogenesis of inflammatory and autoimmune diseases. Integrins are a family of adhesion and signaling molecules made up of paired alpha and beta subunits, and the integrin alpha4beta1 plays a prominent role in the trafficking of mononuclear leukocytes. We have previously described the direct interaction of the signaling adaptor molecule paxillin with the cytoplasmic domain of the alpha4 integrin subunit. This interaction is critical for alpha4beta1 integrin dependent cell adhesion under shear flow conditions as it provides a needed connection to the actin cytoskeleton. Furthermore, the alpha4-paxillin interaction is required for effective alpha4beta1 dependent leukocyte migration and does so through the temporal and spatial regulation of the small GTPase Rac. These findings make the alpha4-paxillin interaction a potentially attractive therapeutic target in controlling leukocyte trafficking.

Human kidney proximal tubular epithelial cell line expresses paxillin induced by TGF-?_1 / 第三军医大学学报

Objective To investigate the expression of Paxillin (Pax) in human kidney proximal tubular epithelial cell line (HKC) induced by transforming growth factor ?1(TGF?1). Methods HKC cells cultured in vitro were divided into three groups at random: control group(C): cultured with free serum medium(FSM), TGF?1-treated groups (T1 and T2): cultured with FSM containing different concentrations of TGF?1 (T1: 5 ng/ml, T2: 10 ng/ml). After 48-hour treatment, the expression of Pax mRNA and protein was detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical staining in HKC cells respectively. Results In Group C, the expression of Pax mRNA and protein in HKC cells was elementary. However, in T1 and T2 groups , the expression of Pax mRNA and protein in HKC cells was greatly increased, especially in T2 group which was more significantly increased than T1 group (P

ATP-induced focal adhesion kinase activity is negatively modulated by phospholipase D2 in PC12 cells

Extracellular ATP has been known to modulate various cellular responses including mitogenesis, secretion and morphogenic activity in neuronal cells. In the ATP-induced morphogenic activity, focal adhesion kinase(s) such as Fak have been suggested to play a critical role. Binding of ATP to its specific cell surface receptor in PC12 cells induces phospholipase D (PLD) activity. However, the role of PLD on ATP-induced Fak activation in PC12 cells remains unclear. In this study, we investigated the role of PLD on the ATP-induced Fak activation and paxillin phosphorylation using two established cell lines: wild type PLD2- and lipase-inactive mutant PLD2-inducible PC12 cells. Stimulation of cells with ATP caused PLD2 activation via classical protein kinase C activation. ATP also induced Fak activation, and paxillin phosphorylation, and were dramatically reduced by wild type PLD2 overexpression but not by lipase-inactive mutant PLD2 overexpression. When the PC12 cells were pretreated with propranolol, a specific inhibitor for phosphatidic acid phosphohydrolase resulting in the accumulation of PA, ATP-induced Fak activation and paxillin phosphorylation were also reduced. We found that inhibition of tyrosine phosphatases by pervanadate completely blocked PLD2-dependent Fak and paxillin dephosphorylation. Taken together, we suggest that PLD2 activity might play a negative role in ATP-induced Fak and paxillin phosphorylation possibly through tyrosine phosphatases.

Tyrosine phosphorylation of paxillin may be involved in vascular smooth muscle contraction

Paxillin is a regulatory component of the complex of cytoskeletal proteins that link the actin cytoskeleton to the plasma membrane. However, the role of paxillin during smooth muscle contraction is unclear. We investigated a possible role for the membrane-associated dense plaque protein paxillin in the regulation of contraction in rat aortic vascular smooth muscle. The tyrosine phosphorylation of paxillin, which was increased by norepinephrine, reached a peak level after 1 min stimulation and then decreased with time. However, norepinephrine induced a sustained contraction that reached a steady state 30 min after application. Pretreatment with tyrphostin, an inhibitor of tyrosine kinase, inhibited the tyrosine phosphorylation of paxillin and also the contraction stimulated by norepinephrine. Both inhibitions were concentration-dependent, and the degree of correlation between them was high. These results show that, in rat aortic smooth muscle, tyrosine kinase(s) activated by norepinephrine may phosphorylate the tyrosine residues of paxillin, thereby providing a source of regulation during vascular smooth muscle contraction.

Expression of p130Cas and paxillin in human breast carcinoma / 中国普通外科杂志

Objective To explore the expression of p130Cas and paxillin in human breast carcinoma and to investigate the relationships of p130Cas and paxillin levels with clinical and pathological characteristics.Methods SP immunohistochemistry staining was applied to detect the expression of p130Cas and paxillin in tumor tissues from 53 cases of primary breast carcinoma,10 cases of breast fibroadenoma and in 10 cases of normal breast tissues.Results Breast carcinoma tissues showed higher levels of p130Cas(P

The effect of TGF-?_1 on expression of paxillin in human renal proximal tubular epithelial cell line / 解放军医学杂志

Objective To explore the effect of transforming growth factor ?_1 (TGF-?_1) on expression of paxillin(Pax) in human kidney proximal tubular epithelial cell line (HKC cells) in different phases. Methods HKC cells cultured in vitro were divided into four groups at random. Control group(C group): HKC cells were cultured with serum free medium (FSM), and TGF-?_1-treated groups (T1, T2, T3 group): HKC cells were cultured with FSM containing 10ng/ml TGF-?_1. In the latter groups, duration of treatment varied as follows: T1 group for 24h, T2 group for 48h, T3 group for 72h. The proliferation of HKC cells induced by TGF-?_1 was assessed by MTT and the expression of Pax was determined. The gene expression of Pax was determined by reverse transcripton polymerase chain reaction (RT-PCR) and the protein expression of Pax was assessed by immunohistochemical staining and Western blot. Results TGF-?_1 could induce the proliferation of HKC cells in a time-dependent manner. In C group there was expression of Pax mRNA and protein in HKC cells. The expression of Pax mRNA and protein in HKC cells was positive in all the T groups, especially in T2 group and T3 group they were significantly increased compared with T1 group (P