1. The Influence On The Mice Germline Mutation Of ESTR By Subcnronically Exposed To Indoor-air In A Newly Decorated Room 2. JWA Functionally Regulates The Migration Of MCF-7 Induced By MIP-1α

(1) The influence on the mice germline mutation of ESTR bysubcnronically exposed to indoor-air in a newly decorated roomIn the recent years, the contaminations of chemical, physical andbiological were shown to be increasing in the indoor-air of China. Since thesynthetic materials and chemical products have been extensively used inbuilding decoration, the effects of the indoor-air quality on health has becomean important concerning. As a result, the concentration of volatile organiccompounds (VOCs), such as formaldehyde, was elevated in most newlydecorated buildings. The VOCs not only contaminate the indoor-air, but alsoobviously harmful to the health of the workers and the residents withoutawareness. The rodent expanded simple tandem repeat (ESTR) DNA consistsof 4- to 6-bp repeat units in long tandem arrays that are unstable in thegermline, tend to mutate by insertion or deletion of a number of repeat units,and has provided an effective genetic tool for detecting induced mutations inlab mice at lower treatment doses and smaller sample sizes than ever before.Here, we established two kinds of animal models to test ESTR germlinemutation rates in ICR laboratory mice chronic as exposed to indoor-air at anewly decorated room, and also in mice acute inhalation of FA (formaldehyde),with those exposed at an animal care facility location as control. The results of the concentions of TVOC, FA, benzene and ammonia of theindoor-air on the first day the ICR mice exposed to were 12, 3, 3.6, 3.18 timeshigher than the control site and 3.6, 22, 6.75, 1.75 times higher than indoor-air-quality (IAQ) related air quality standards in China respectively. Theconcentration of FA in the exposure room fell from 0.22 mg/m~3 to 0.084 mg/m~3during the 10 weeks exposure time. Body mass and general growth conditionsof the animals did not show significant differences among the three groupsfrom the first day of exposure up to 16 weeks.Single ESTR probe Ms6-hm detected significant elevations in germlinemutation frequency in the indoor-air exposure group and FA acute inhalationgroup over the control group, respectively. Furtehr analysis showed that thepaternal mutations of the two exposure groups were 2.0 and 1.9 times morefrequent than the control mice, which make up most of the overall elevation inthe exposed groups. Maternal mutation rates showed no significant defferencebetween the two groups and the control, respectively.Taken together, the germline mutations of Ms6-hm in both chronicexposure and FA acute inhalation were elevated compared with the control,respectively. Premeiotic male germ cells are sensitive to indoor-air pollutantsof newly decorated room and FA acute inhalation. According to our findings,these populations may be at risk of increased heritable mutation frequencythrough their exposed fathers. Obviously, further investigations are necessarynot only to understand the inherent relationships between germline mutationand indoor-air pollution, but also further reinforce IAQ management andpollution control.2. JWA functionally regulates the migration of MCF-7 induced by MIP-1αSeveral studies reported that some tumor cells such as MCF-7 humanbreast carcinoma cells expressed chemokine receptors, which possessedproperties that may be associated with migration and metastatic capacity. Chemokines and their receptors play important roles in acute and chronicinflammation, angiogenesis, immunological regulation, anti tumor and antiHIV infection, etc. Recently, studies have demonstrated that JM4, amembrane-associated protein, was found to be co-precipitated with CCR5, andit was suggested to function in trafficking and membrane localization for theCCR5 receptor. As a ubiquitously expressed protein, JM4 was reported toshare about 62 % sequence similarity with JWA (BenBank: AF070523), anovel cytoskeleton associated protein, was primarily cloned from humantracheal bronchial epithelial (HBE) cells. Mitogen -activated protein kinase(MAPK) families were implicated to be involved in cellular proliferation,differentiation, apoptosis and migration processes. Further studies showedJWA might function as a putative MAPK activated protien. As the migrationof MCF-7 cells might be induced by MIP-1α, the aim of this study was todetermine the role of JWA in MIP-1αinduced MCF-7 breast carcinoma cellmigration and the potential molecular mechanisms.The interaction of CCR5 and JWA in MCF-7 cells was confirmed first atendogenous level. The confocal laser scanning microscopy assays also showedthat both JWA and CCR5 were distributed similarly and intracellularco-localized in MCF-7 cells. After the treatment of MCF-7 cells with MIP-1α,up-regulated JWA protein and CCR5 receptor expression levels were observed.The further studies were designed to understand if JWA involves in theregulation of MIP-1αinduced cell migration. Results showed that after 24 hincubation, 100 ng/ml MIP-1 a induced a most significant migration in MCF-7breast carcinoma cells. In JWA-deficient MCF-7 cells, the activation ofMEK-ERK1/2 signal pathway was significantly blocked and the migration ofMCF-7 cell induced by MIP-1αwas also inhibited. After recovering theexpression of JWA protein by transfection of JWA express vector (rescueassay), the functions of MIP-1αin inducing cell migration were also recovered.Interestingly, once the two serines phosphorylation motifs in JWA protein were mutated, the functions of MIP-1αwere unrecoverable.Data in the present study demonstrated that JWA interacts with CCR5,and the expression levels of these two proteins are significantly induced byMIP-1αtreatment. The accelerated migration of MCF-7 cells induced by MIP-1αis mainly via the CCR5 receptor linked MEK-ERK1/2 signal pathwaywhich is dependent on the expression and activation of JWA. Eitherknock-down JWA or mutate JWA PKA/PKC phosphorylation motifssignificantly blocked cell migration in response to MIP-1α. Data indicate thatJWA protein and CCR5 are likely to be important targets for the developmentof inhibitors with potential application in breast cancer therapies.