Functional characterization of breast cancer using pathway profiles

BACKGROUND: The molecular characteristics of human diseases are often represented by a list of genes termed “signature genes”. A significant challenge facing this approach is that of reproducibility: signatures developed on a set of patients may fail to perform well on different sets of patients. As diseases are resulted from perturbed cellular functions, irrespective of the particular genes that contribute to the function, it may be more appropriate to characterize diseases based on these perturbed cellular functions. METHODS: We proposed a profile-based approach to characterize a disease using a binary vector whose elements indicate whether a given function is perturbed based on the enrichment analysis of expression data between normal and tumor tissues. Using breast cancer and its four primary clinically relevant subtypes as examples, this approach is evaluated based on the reproducibility, accuracy and resolution of the resulting pathway profiles. RESULTS: Pathway profiles for breast cancer and its subtypes are constructed based on data obtained from microarray and RNA-Seq data sets provided by The Cancer Genome Atlas (TCGA), and an additional microarray data set provided by The European Genome-phenome Archive (EGA). An average reproducibility of 68% is achieved between different data sets (TCGA microarray vs. EGA microarray data) and 67% average reproducibility is achieved between different technologies (TCGA microarray vs. TCGA RNA-Seq data). Among the enriched pathways, 74% of them are known to be associated with breast cancer or other cancers. About 40% of the identified pathways are enriched in all four subtypes, with 4, 2, 4, and 7 pathways enriched only in luminal A, luminal B, triple-negative, and HER2+ subtypes, respectively. Comparison of profiles between subtypes, as well as other diseases, shows that luminal A and luminal B subtypes are more similar to the HER2+ subtype than to the triple-negative subtype, and subtypes of breast cancer are more likely to be closer to each other than to other diseases. CONCLUSIONS: Our results demonstrate that pathway profiles can successfully characterize both common and distinct functional characteristics of four subtypes of breast cancer and other related diseases, with acceptable reproducibility, high accuracy and reasonable resolution

OR-036 Aerobic exercise activates CHI3L1/PAR2 to promote cardiomyocyte proliferation and protect cardiac function in rats with MI

Objective Objectives: The aim of this study is to investigate the changes of protein expression of chitinase-3-like protein 1 (CHI3L1) and its receptor PAR2 (protease-activated receptor 2) after exercise, and the possible mechanism to promote the proliferation of cardiomyocytes and protect MI rats heart.Methods Methods: Using rhCHI3L1 (150 ng/ml), AMPK agonists (AICAR, 50 mM) separately or together administer stimulation for 24 hours.H9C2 cells were divided into control group (H9C2 group), CHI3L1 recombinant protein intervention group (H9C2+rhCHHI3L1), AMPK agonist intervention group (H9C2+AICAR), and CHI3L1 recombinant protein and AMPK agonist combined intervention group (H9C2+rhCHI3L1+AICAR).SD rats were subjected to left anterior descending(LAD) coronary artery ligation to prepare MI models and randomly divided into sham operation group (S), myocardial infarction group (MI), myocardial infarction aerobic exercise group (ME), and injection of PAR-2 blocker agent (FSLLRY-amide) was divided into 7-day PAR2 blocker injection group (7d+FS), MI 7-day saline injection group (7d+SA), MI 14 days PAR2 blocker injection group (14d+FS), MI 14-day saline injection group (14d+SA). One week after surgery, the ME group were subjected to oneweekadaptively exercise followed by four weeks aerobic exercise.At the end of the training, the rats were intraperitoneally anesthetized the next day. Hemodynamic measurements of LVEDP, ±dp/dt max, and LVSP were used to evaluate cardiac function.The protein levels of CHI3L1/PAR2, pPI3K/PI3K, pAKT/AKT, pERK/ERK and Cyclin D1 in H9C2 and rat hearts were determined by Western blotting. The proliferation of H9C2 was detected by CCK-8. The proliferation of H9C2 and cardiac tissue was observed by immunofluorescence. Masson staining was used to observe myocardial collagen volume percent (CFV%).Results Results: Compared with H9C2 control group, the expression of CHI3L1, pPI3K/PI3K, pAKT/AKT, pERK/ERK, and Cyclin D1 protein increased significantly after rhCHI3L1 and AICAR intervention for 24 h, respectively. CCK-8 test and immunofluorescence indicated that H9C2 had significant proliferation effect. In MI rat heart, compared with MI group, the expression of CHI3L1, pPI3K/PI3K, pAKT/AKT, pERK/ERK and Cyclin D1 protein in ME group increased significantly, the number of cell proliferation increased, and LVEDP significantly decreased, ±dp/dt max, LVSP significantly increased, and the CFV% decreased significantly.Conclusions Conclusion: Exercise may promote cardiomyocyte proliferation and improve cardiac function in MI rats by activating the CHI3L1/PAR2-PI3K-AKT-ERK signaling pathway

PO-190 Exercise Training Protects Against Cardiac Pathological Remodeling in Myocardial Infarction rats via Improving Mitochondrial Biogenesis

Objective  Growing evidence suggests that exercise training reverses cardiac pathological remodeling and cardiac dysfunction during myocardial infarction (MI), but the underlying mechanisms have not been fully understood. In this study, we investigated the impact of exercise training on cardiac function, myocardial fibrosis, apoptosis, oxidative stress and mitochondrial biogenesis. Methods Sprague Dawley rats were subjected to MI by permanent ligation of the left anterior descending (LAD) coronary artery or Sham operation. Rats with MI were randomly assigned to sedentary MI group (MI) and MI with exercise training group (MI+EX), and compared to sham-operated group (Sham). Haemodynamics and Masson staining were conducted to evaluate the effect of exercise training on cardiac function and myocardial fibrosis. Myocardial apoptosis, oxidative stress, mitochondrial biogenesis and molecular signaling mechanism were analyzed. Results  Exercise training significantly improves cardiac function and mitigates the MI-induced cardiac pathological remodeling. Meanwhile, Exercise training significantly attenuates MI-induced apoptosis, oxidative stress and mitochondrial biogenesis. In addition, activation of PI3K pathway following MI is further induced by exercise training. Conclusions  Exercise training protects against MI-induced cardiac dysfunction and pathological remodeling through preventing myocardial apoptosis and oxidative stress, and enhancing mitochondrial biogenesis

PO-163 Aerobic exercise activates myocardial FGF21/FGFR1/PI3K-AKT signaling pathway and inhibits cardiomyocyte apoptosis in post-myocardial infarction rats

Objective To investigate the effect of aerobic exercise on the expression of fibroblast growth factor 21 (FGF21) and cardiomyocyte apoptosis in Myocardial Infarction (MI) rats. Methods male SD rats were randomly divided into three groups：the sham operation (S), sedentary MI group (MI) and MI with aerobic exercise group (ME). The MI model was established by ligation of the left anterior descending branch of the left coronary artery. ME group were trained four weeks after the operation. LVSP, LVEDP and ±dp/dtmax were used to evaluate cardiac function. H9C2 cardiomyocytes were stimulated by 400 μmol/L H2O2 for 4h to simulate myocardial apoptosis mode. AMPK agonist AICAR and FGF21 receptor inhibitor PD166866 were used to interfere with H9C2. Myocardial collagen volume fraction was calculated by Masson staining and myocardium FGF21, FGFR1, Bax, Bcl-2 and PI3K-AKT pathway by western blotting or RT-Qpcr. Cardiomyocytes apoptosis was evaluated by TUNEL. Results Compared with S, the expression of FGF21, FGFR1, Bax, Bcl-2 and PI3K, AKT increased significantly in MI group, the apoptotic cardiomyocytes and collagen fibers increased significantly, but the cardiac function decreased. Compared to MI, myocardium FGF21, FGFR1 and PI3K, AKT were further increased in ME group, the Bax/Bcl-2 and the apoptotic cardiomyocytes decreased significantly. The percentage of collagen fibers decreased and the cardiac function was improved. Myocardium FGF21 was positively correlated with the inhibition of cardiomyocyte apoptosis and the improvement of cardiac function. Furthermore, the expression of Bax/Bcl-2, TNF-α/IL-10 and the apoptotic cardiomyocytes was significantly increased by PD166866, but PI3K-AKT pathway decreased significantly by PD166866. However, AICAR single intervention or PD166866 simultaneous intervention also can reverse this adverse effects. Conclusions Exercise can increase myocardial FGF21/FGFR1 with MI. The one of the mechanisms is to activate PI3K-AKT pathway to inhibit cardiaomyocyte apoptosis and inflammatory. It indicates that FGF21/FGFR1/PI3K-AKT signaling pathway plays an important role in inhibiting myocardial apoptosis and improving cardiac function

Static and dynamic performance evaluation of a 3-DOF spindle head using CAD–CAE integration methodology

Accurate and rapid modeling and performance evaluation over the entire workspace is a crucially important issue in the design optimization of parallel kinematic machines (PKMs), especially for those dedicated for high-speed machining where high rigidity and high dynamics are the essential requirements. By taking a 3-DOF spindle head named A3 head as an example, this paper presents a feature-based CAD–CAE integration methodology for the static and dynamic analyses of PKMs. The approach can be implemented by four steps: (1) creation of a parameterized geometric (CAD) model with analysis features in SolidWorks; (2) extraction of the features from the CAD model using the Application Programming Interface (API) available in SolidWorks; (3) formulation of a CAD model in SAMCEF by mapping the configuration features from SolidWorks to SAMCEF; and (4) conversion of the analysis features into a scripting language named Bacon for Finite Element Analysis (FEA). The merit of this approach lies in that the FE model at different configurations can be updated automatically in batch mode, and PKMs having different topologies can be modeled with ease thanks to the down to link/joint level featuring. The experiment is also carried out to verify the effectiveness of the proposed approach

PO-167 Effects of Hydrogen-rich Water on Rat Skeletal Muscles' Oxidative Damage and Autophagy Induced by Repeated Exhaustive Exercise

Objective This study was carried out to detect the effects of hydrogen-water intervention on oxidative stress and cell autophagy in skeletal muscle of rats with repeated exhaustion.  Methods 30 male SD rats in the age of 3 months, weighing 180-210g, were randomly divided into control group (C), repeated exhaustion group (EX), and the repeated exhaustion with hydrogen intervention group (EH), 10 rats in each group. The EX and EH groups were subjected to a four-weeks of repeated-exhaustive exercise. The initial speed of the exercise was 15 m/min, and increasedby 5 m/min every 5 min, the final speed is 35m/min until  exhaustion, 5 d/wk, with a total of 4 wk. In EH group, hydro-water was given to rats 30 mins before exercise. The ultrastructural changes of skeletal muscle were observed by using a transmission electron microscopy. Activity of SDH and CK in serum and SOD activity, MDA content and T-AOC level in skeletal muscle tissue were detected. Western blotting was used to detect the proteins expression of autophagy related proteins in skeletal muscle, mTOR, p-mTOR, LC3B-2 and P53.  Results Compared with the EX group, in the EH group, the ultrastructural damage and mitochondrial swelling were significantly reduced, and the time of exhaustion was significantly prolonged (p<0.05), Serum SDH activity increased significantly (p<0.05), CK activity decreased significantly (p<0.05), and skeletal muscle tissue SOD activity and total antioxidant capacity significantly increased (p<0.05), MDA content decreased significantly (p<0.01), mTOR and p-mTOR protein expression was significantly increased(p<0.05), the LC3B-2 and P53 protein expression was significantly lower (p<0.05). Conclusions Hydrogen water intervention could significantly improve repeatedly exhaustion exercise result in rat skeletal muscle injury, oxidative stress and cell ultrastructure damage excessive autophagy, improving oxidation resistance of the skeletal muscle and exercise endurance

PO-144 Intermittent Exercise Activates NRG1-SERCA2a Pathway to Improve Cardiac Function in Myocardial Infarction Rats

Objective  Intermittent exercise can improve cardiac function in rats with myocardial infarction. The Neuregulin-1(NRG1)/SERCA2a palys a critical role in maintain cardiac function. We want to investigate the effect of Neuregulin-1 (NRG1) on NRG1-SERCA2a signaling pathway activated by intermittent exercise and on improves cardiac function in rats with MI. Methods 32 male sprague-dawley rats were randomly divided into four groups (n=8): Sham-operated group (S), sedentary MI group (MI), MI with interval training group (ME), ME with inhibitor AG1478 group (MA). ME and MA model after the MI model was established by ligation of the left anterior descending coronary artery, and began training 1 week after MI surgery. The S model only by threading without ligation. Rats in ME and MA model taken one week adaptive training, then began 8-week interval training. MA model were injected with inhibitor AG1478, once every two days. The 24h after training, rats were anesthetized, the LVSP, LVEDP, ±dp/dt max were tested by carotid artery intubation which in order to evaluate cardiac function. The protein expression of NRG1, PI3K, Akt, eNOS, PKG, PLN, SERCA2a in myocardium were measured by Westernblotting, themRNA expression of serca2a were tested by RT-qPCR. Results Compared with S, the protein expression of NRG1, PKG, peNOS, pAkt, pPLN, pPI3K and SERCA2a decreased, serca2a mRNA expression decreased, LVSP and ±dp/dt max significantly decreased, LVEDP significantly increased; Compared with MI, the protein expression of NRG1, PKG, peNOS, pAkt, pPLN, pPI3K and SERCA2a increased, serca2a mRNA expression increased, LVSP and ±dp/dt max significantly increased, LVEDP decreased, and the effect of exercise were weaken by inhibitor AG1478. Correlation analysis showed that the myocardial pPLN and SERCA2a protein expression both were positively correlated with LVSP, ±dp/dtmax, and negatively correlated with LVEDP. Conclusions Intermittent exercise can increased myocardial NRG1 protein expression and activates NRG1-SERCA2a signaling pathway, improve myocardial infarction cardiac function

PL - 027 Up-regulation of NRG1 improves cardiac repair in zebra fish and involved in the cardioprotective effects of exercise training in rats of myocardial infarction

Objective Myocardial infarction (MI) remains a leading cause of morbidity and mortality worldwide. Exercise training could improve cardiac function following MI. However, the mechanisms are still not well-known. Neuregulin 1 (NRG1)plays an important role in heart development and regeneration.In this study, we investigated the effect of NRG1 on cardiac regeneration in a zebrafish model, detected whether exercise could improve cardiac function through regulating NRG1 expression in infarcted heart and explore the possible role of up-regulation of NRG1 in skeletal muscle play in the cardioprotective effects in rats with MI. Methods Transgenic zebrafish line, cmlc2:CreERandβ-act2:BSNrg1,wereusedto study the effect of NRG1 on heart growth and regeneration after injury. PCNA was detected by immunofluorescence staining andmRNAexpression of gata4, nkx2.5, tbx5, smyd1b, hsp90α and murf were tested by RT-PCR.Sprague-Dawley rats were used to establish MI model and underwent fourweeks of exercise training (ET) or pAAV-{dMCK promoter}rNRG1-eGFP intervention.AG1478 was used asan inhibitor of NRG1/ErbBs signaling pathway. Cardiac function and structure,cardiomyocyte proliferation and NRG1 expression were detected in the heart or skeletal. Results Cardiac-specific overexpression of NRG1 induced cardiac hypertrophy and cardiomyocyte proliferation, regulated the mRNA expression of gata4, nkx2.5, tbx5, smyd1b, hsp90α andmurf in uninjuriedzebrafish, and promote cardiac repair and regeneration after injury in the zebrafish.Exercise activated NRG1/ErbBs signaling pathway, improved cardiac remodeling and heart function, enhanced cardiomyocyte proliferation, reduced cardiomyocyte apoptosis, ROS level and MuRF1 protein expression in rats with MI. BlockingErbB signaling attenuated the ET-induced cardioprotection effects in rat with MI.up-regulation of NRG1 expression in skeletal muscle could increase the protein level of NRG1 in serum and infarcted heart, improve cardiomyocyte proliferation and reduce the level of cardiac fibrosis, finally promote cardiac function. Conclusions Up-regulation of NRG1 expression in the heart or skeletal musclemay be one of the underlying mechanisms of thebeneficial effects of exercise training following MI

PO-132 Exogenous VD3 alleviates chronic fatigue syndrome by activating MEKs/ERKs-SIRT1 signaling pathway in skeletal muscle

Objective To investigate the effects of exogenous VD3 on exercise-induced chronic fatigue syndrome (CFS) and its mechanism. Methods 80 male mice were randomly divided into 5 groups（n=16）. One group was named as quiet control group (C) while the other four were used to build CFS model by forced swimming. Among the four groups, three were selected for feeding VD3 1 h before exercise (bVD), 15 min after starting exercise (mVD) and 1 h after exercise (aVD) respectively, and the group without VD3 feeding was CFS model group (CFS). The behavior and condition of mice was observed during the whole process. After the training was completed totally, the tail suspension test and the forced swimming test were implemented, and then the blood and quadriceps were dissected. Serum testosterone and corticosterone content, as well as the antioxidant system indexes in the quadriceps, including malondialdehyde (MDA), total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-PX) and total superoxide dismutase (T-SOD), were measured. Routine western blotting experimentation was used to measure signal pathway indicators, containing SIRT1, SIRT3, ERK1/2, p-ERK1/2 and MEK1/2. Results Compared with C group, CFS group appeared serious damage caused by fighting, and the concentration of serum testosterone decreased significantly (p<0.01 or p<0.05) while cortisol concentration increased significantly (p<0.01 or p<0.05). Regarding antioxidant stress system indexes, the expression of MDA, T-SOD and GSH-PX significantly increased (p<0.05). The concentration of MEK and SIRT3 decreased significantly (p<0.01 or p<0.05). Compared with CFS group, VD intervention group (bVD, mVD and aVD) showed less damage caused by fighting and significantly lighter body weight (p<0.05), and the concentration of serum testosterone increased significantly (p<0.05) while that of cortisol decreased significantly (p<0.01 or p<0.05). The expression of MDA decreased significantly (p<0.05), on the countrary, T-AOC, T-SOD and GSH-PX increased significantly (p<0.05). The expressions of MEK, p-ERK/ERK, SIRT1 and SIRT3 were significantly upregulated (p<0.01 or p<0.05). Conclusions Exogenous VD3 could alleviate CFS, which probably related to activate the MEKs/ERKs-SIRT1 signal transduction axis and hence regulate the expression of SIRT3

PO-169 Effect of Exercise-Induced Skeletal Muscle FSTL1 on Cardiac Structure and Function in Myocardial Infarction Rats

Objective The aim of this study is to investigate the effect of skeletal muscle-derived FSTL1 on cardioprotection in myocardial infarction rats after resistance exercise or tibialis anterior muscle injection of follistatin-like protein 1 (FSTL1) adeno-associated virus vector and its possible signaling mechanisms. Methods The male Sprague-Dawley rats were randomly divided into five groups (n=10): Sham-operated group(S), sedentary MI group (MI), MI with resistance exercise group (MR), MI with empty adeno-associated virus (AAV) vector group (MV) and MI with FSTL1-AAV group (MF) after the MI model established which was induced by left anterior descending (LAD) coronary artery ligation. S group underwent threading without ligation. 1 week post MI, rats in MR group underwent resistance exercise for 4 weeks, rats in MV and MF group were injected AAV empty vector and FSTL1-AAV in the tibialis anterior muscle of the left limb, respectively. The next day after exercise, rats were anesthetized and heart function was measured. Collagen volume fraction(%) of myocardium were observed and calculated by Masson staining; cardiomyocyte proliferation was measured by immunofluorescence; cardiomyocyte apoptosis was detected by TUNEL staining; The protein expression of skeletal muscle and serum FSTL1 and myocardium FSTL1, DIP2A, pAkt/Akt, p-mTOR/mTOR, CyclinD1, CDK4 and Bcl2/Bax in myocardium were measured by Western blotting. Results The skeletal muscle FSTL1 protein expression was decreased but the serum and myocardium FSTL1 were upregulated in MI group. The myocardium fibrosis, cardiomyocyte proliferation and cardiomyocyte apoptosis were increased and the heart function was declined after MI. After resistance exercise or tibialis anterior muscle injection of FSTL1-AAV, the skeletal muscle, serum and myocardium FSTL1 protein expression were significantly increased, and there was a significant positive correlation between each data. Myocardium fibrosis and cardiomyocyte apoptosis were also decreased, cardiomyocyte proliferation was increased and the heart function was significantly improved after FSTL1-AAV injection. Conclusions Resistance exercise increases skeletal muscle FSTL1 expression. Skeletal muscle-derived FSTL1 can reach the heart through blood circulation, promote cardiomyocyte proliferation, inhibit cardiomyocyte apoptosis, reduce myocardium fibrosis and improve heart function in MI rats. Myocardium FSTL1 binds to its receptor, DIP2A, and activates the Akt-mTOR signaling pathway might be the potential mechanism of this protective effect