MCF7 HER2 tumors were much more sensitive to gefitinib and RAD001 than JIMT 1. Rising the gefitinib dose to 200 mg/kg and RAD001 over 2. 5 mg/ kg resulted inside a higher therapeutic impact represented by stable condition in lieu of tumor regression in animals bearing MCF7 HER2 tumors. Gefitinib used at one hundred mg/kg and RAD001 employed at 1. 75 mg/kg diminished tumor volume by two. seven fold and one. 6 fold, respectively, relative on the automobile management group but these variations weren’t statistically important.

Having said that, the common MCF7 HER2 tumor volume on the final day of therapy while in the mixture inhibitor,modulator,library treated group was signifi cantly smaller sized than during the manage or RAD001 group. In contrast, the main difference concerning the combination and gefitinib handled tumors was not statistically major. These data demonstrate the combination therapy was a lot more potent compared to the single medication when in contrast to automobile taken care of controls. Importantly, the combination prevented even more development of TZ delicate and resistant tumors. The synergy analy sis based around the median impact methodology developed by Chou and Talalay couldn’t be performed around the in vivo information mainly because the combination was only examined at 1 dose of gefitinib.

It needs to be mentioned that none with the therapy regi mens triggered any considerable physique bodyweight reduction in ani mals. Comprehensive animal health monitoring data suggested that gefitinib and RAD001 were nicely tolerated with the doses made use of, whether the medication have been made use of alone or in combination. It’s crucial to note that we also tested sensitivity of JIMT one tumors to TZ in Rag2M mice. The outcomes of this review presented in Further selleck file 1 show that remedy with TZ over the course of 27 days did not result in inhibition of tumor volume, thus, confirming the resistance of JIMT 1 cells to TZ, as previously established by many others.

Effects of gefitinib, RAD001 along with the blend on tumor tissue characteristics Immunohistochemistry based mostly tumor tissue map ping methods have been used to investigate adjustments in JIMT 1 tumors harvested from animals treated for 28 days with 100 mg/kg gefitinib, one. 25 mg/kg RAD001 or the gefitinib and RAD001 mixture and in MCF7 HER2 tumors harvested from animals handled for 25 days with one hundred mg/kg gefitinib, one. 75 mg/kg RAD001 or even the blend. The area of confluent TUNEL optimistic tissue, herein described as necrosis and TUNEL staining inside areas of viable tumor selleck chemical tissue, indicative of apoptotic cells, in conjunction with CD31 staining and proliferation status of tumor tissue were assessed.

The outcomes indicate the imply degree of necrosis and apoptosis didn’t differ involving therapy groups in JIMT 1 and MCF7 HER2 tumors. Since gefitinib and RAD001 happen to be reported to exert anti angiogenic effects, we also investigated possible modifications in tumor vascularization. An general larger ves sel density was noticed within the MCF7 HER2 tumors in which the median distance of tumor tissue towards the nearest CD31 optimistic object was half that from the JIMT one tumors. The median dis tance of tumor tissue towards the nearest CD31 favourable ves sel in JIMT one tumors derived from animals handled with gefitinib was considerably decreased in contrast to car manage suggesting a rise in vasculariza tion. No alterations had been noticed in tumors derived from animals treated with RAD001 alone and the blend for that most part reflected the results of gefitinib.

Numerically, we are able to calculate the inter bin sensitivity error working with the next equation is in opposition to tumor suppressors, proteins that have been shown to possess a unfavorable correlation with the devel opment of cancer. To capture the conduct of oncogenes, we partially formulate our dilemma on two rulesRule 1 basically says that if inhibiting a number of target proteins has blocked signaling pathways, then inhibiting more target proteins won’t open any path which has already been blocked. Rule two captures the truth that if a set of target protein inhibitors is unsuccessful in blocking the paths of the circuit, then any diminished quantity of target professional tein inhibitors amongst the inhibiting proteins can’t block all the paths.
The over guidelines presume the kinases in emphasis are oncogenes, genes that encourage PF-04929113 price cancer development and whose inhibition can prevent tumor improvement. Nearly all kinases from the Drug Display panel behave as oncogenes, and as such, our method utilizes the above principles. Target sets leading to mixture scores that don’t follow the rule based behavior incur an inconsistency penalty. This penalty is calculated as follows the place ? would be the indicator perform which can be one when the experimental drug score is inconsistent with the predicted subsetsuperset bin score. We now current the finish target set score, and as such, the equation that we wish to resolve This evaluation has a single notable flaw if we attempt selleck chemical to only separate the many medication into bins based on inter bin sensitivity error, we will make an more than fitted alternative by breaking every single drug into a person bin.
We get two ways to prevent this. 1st, we attempt to minimize the amount of targets during construction of T0. 2nd, we which reduces for the minimization issue we wish to fix integrate an inconsistency phrase to account for target conduct that we take into consideration to get biologically inaccurate. To expand about the above point, we consider you’ll find two complementary guidelines by which kinase targets behave. Exploration has shown that the bulk of viable kinase tar gets behave as tumor promoters, proteins whose presence and lack of inhibition is connected for the continued survival and growth of a cancerous tumor. These targets in essence have a optimistic correlation with cancer progression. This For brevity, we will denote the scoring function of the target set with respect to your binarized EC50 values S as well as scaled sensitivity scores Y. Since the S and Y sets are going to be fixed when target set generation commences, we lessen this notation even more to. Note that T ? K exactly where K denotes the set of all attainable targets. two|K| could be the complete amount of prospects for T that is really large and thus prohibits exhaustive search.

Then, the media was modified plus the cells had been cul tured for two days from the presence of serum. We identified that micromolar concentrations had been cytotoxic, since cell numbers decreased after two days, whereas nanomolar concentrations were growth inhibitory. Melanoma cells showed dose dependent inhibition with 0. 01 nM to one hundred nM of BAY43 9006, or rapamycin. Proliferation with the cells was inhibited in both 5% or 0. 5% serum. Amid the melanoma cell lines, there was a significant distinction within the volume of inhibition at ten nM BAY43 9006 or rapamycin. We observed that melanoma cell lines that have the V599E mutation in B Raf have been far more delicate to BAY43 9006 and also to rapamycin, compared to cell lines with wild sort B Raf. This variation in development inhibition was observed in two extra cell lines, one particular wild kind and one V599E.
Thus, nanomo lar concentrations of either BAY43 9006 or rapamycin inhibit the proliferation of melanoma cells, whether they’ve mutated B MLN2480 Raf. Combining Rapamycin with BAY43 9006 synergistically inhibits serum dependent proliferation of melanoma cells Melanoma cell proliferation was inhibited by either BAY43 9006 or rapamycin in excess of the 0. 01 one hundred nM con centration selection. A mixture of your two medicines was markedly a lot more productive than either drug alone at inhibit ing serum stimulated melanoma cell proliferation. For example, 0. 01 nM of each drug together was far more effec tive at inhibiting melanoma cell proliferation than one nM of either drug alone. To assess synergism versus additivity quantitatively, we used a focused isobologram strategy.
Remedy read review of three melanoma cell lines with rapamycin alone induced a 70% growth inhibition from approxi mately ten nM to 2 nM. These were plotted within the ordinate. The IC70 concentra tion for BAY43 9006 alone was within the choice of approxi mately 5 to 10 nM, in numerous cell lines, and these were plotted about the abscissa. In contrast on the single agents, the IC70 for your dose pairs falls beneath the line, for every of those melanoma cell lines, indicating the combination is synergistic. On top of that, VMM18, which includes the V599E substitution, was far more delicate towards the combina tion treatment method than melanoma cell lines with wild style B Raf, consistent together with the enhanced sensitivity with the 10 nM dose of each agent.
On the other hand, all melanoma cell lines tested displayed synergistic inhibi tion of proliferation, indicating that these medication had been more efficient in mixture than alone. Rapamycin and BAY43 9006 inhibit phosphorylation of proteins in the mTOR signaling pathway in melanoma cells Melanoma cells had been treated with rapamycin and BAY 43 9006, either singly or in blend, for 1 hour, and protein phosphorylation was examined by Western blot examination 24 hours later on. Rapamycin is an inhibitor of mTOR kinase and lowers phosphorylation of its sub strates, p70S6K and 4EBP1.

85 two. 81m, ten. 38 one. 52m, ten. 70 2. 35m and 9. 11 2. 44m in SMMC 7721, MHCC97 L, MHCC97 H and HCCLM6 cells, respectively. As expected, SMMC 7721 cells, which have the lowest amounts of pERK, were considerably significantly less sensitive to sorafenib mediated growth inhibition than the other three HCC cell lines with greater basal pERK lev els. Meanwhile, a significant adverse correlation was observed between the IC50 values of sorafenib in these HCC cell lines and their pERK density values, indicating the results of sorafenib on cell pro liferation were drastically correlated with basal pERK levels in these HCC cell lines. Opposite final results had been observed with remedy with the standard chemotherapy drug five FU. five FU inhibited HCC cell proliferation with an IC50 of four. 24 0. 87 mg l, 79. 71 24.
49 mg l, 41. 21 21. fifty five mg l and 187. 45 78. 05 mg l in SMMC 7721, MHCC97 L, MHCC97 H and HCCLM6 cells, respectively, with sizeable statistical variations. The SMMC 7721 cells, with decrease pERK expression, demonstrated a greater sensitivity more helpful hints to five FU. Nevertheless, MHCC97 L, MHCC97 H, and HCCLM6 cells, with larger pERK expression, exhibited much more resistance to this drug. The ultimate inhibition price ahead of reaching a plateau in these three cell lines was about 35%, 40%, and 45%, respectively, just about every in comparison with its handle group. On top of that, a substantial correlation was observed concerning the IC50 values of 5 FU and pERK density values, indicating the resistance to five FU was signifi cantly connected with basal pERK expression in these HCC cell lines.
Results of MEK ERK pathway inhibition and pERK reduction on sensitivity to sorafenib To a lot more straight selleck identify the romance amongst pERK expression and sensitivity to sorafenib, we inhibited the MEK ERK pathway and lowered basal pERK expres sion in MHCC97 H cells via U0126, a selective inhibitor of MEK 1 and MEK two, after which compared cellular responsiveness to sorafenib to that of untreated cells. To prevent achievable direct development inhibition by U0126, expo certain of cells to this drug was restricted to six hrs according to our preliminary experiments. Quantification of cellular pERK ranges by immunocytochemical evaluation indicated that constitutive ERK phosphorylation was strongly decreased in MHCC97 H cells soon after therapy with 20m U0126 for six hours, relative to your degree observed while in the untreated cells, which induced virtually no detectable systemic toxicity on cell proliferation. From the following experiments, we in contrast sorafenib responsiveness of MHCC97 H cells pretreated with 20m U0126 for 6 hrs to an untreated management. Cell viability assay uncovered the pretreated cells were considerably significantly less delicate to sorafenib mediated growth inhibition, with an IC50 of 17.

The Z aspect measures the variation and separation bands of an assay thereby provid ing a statistical measure of its high-quality. It requires into account the dynamic range and variability in the positive and nega tive control measurements. We determined a Z fac tor of 0. 48 employing the ImageXpress Micro and MetaXpress computer software from Molecular Devices and 0. 60 making use of the Cell Lab IC100 and Cytoshop software from Beckman Coulter. Both values are deemed appro priate for a thriving screen. Figure 1g shows the sig nal to background ratios using each imaging platforms. Principal screen So as to assess the utility in the above protocol, we performed a pilot experienced screen of 7368 compounds in duplicates for any total of 44 384 effectively plates. LNCaP S14 cells have been incubated with person com pounds for 18 h, followed by fixation, staining, imaging and analysis as described in Solutions.
We scored the percentage of cells good for p27 in every single effectively relative to the vehicle handle. Figure 2a shows the activity of selleckchem good control com pounds, the adverse control com pound DMSO, the non certain staining and small molecules derived from com pound libraries across all screening plates prior to normalization. Adverse controls gave basal percentages of p27 optimistic cells ranging between 2% and 13% even though the constructive control roscovitine improved these populations up to 50%. As anticipated, the typical of 320 nicely measurements per plate that is, the percentage of p27 positive cells in wells treated with person compounds was comparable to the percentage of p27 optimistic cells in the damaging manage, indicating that most compounds tested had been inactive in causing p27 accumulation.
Normalization of raw information for 7368 compounds was performed by calculating Z scores, which represent the amount of standard deviations by which a signal to get a offered compound differed in the imply signal of your whole plate. It excludes handle measurements alto gether under the assumption that most compounds are inactive and can serve as intrinsic controls. The summed Z scores for one particular set of duplicates showed clear separation amongst the optimistic control compound roscovitine and also the non particular staining. Additionally, most summed Z scores had values comparable to the adverse controls, indicating that we did not incur errors triggered by plate position of compounds. This was additional evaluated by visualizing just about every plate as a heat map of individual Z scores. As shown in the example of a single plate in Figure 2c, potential hits have been randomly distributed more than the plate. The evaluation also identified many false positive compounds with extremely higher Z scores, namely known DNA binding compounds of red colour.

Lastly, the drugs at present utilised for the treatment of OA are aimed at minimizing discomfort and do not possess any illness modifying activity. Studying the synovial fluid proteome should really yield a higher concentration of possible biomarkers than serum or plasma, because the synovial fluid is in direct physical contact together with the synovium, ligament, meniscus, joint capsule and bone. Alterations within the structure and metabolism of any of these tissues in the course of illness should be reflected as al terations inside the composition of the synovial fluid proteome. Thus, the synovial fluid proteome has the prospective to indicate the severity and progression in the illness. Advances in proteomic technologies have facilitated exten sive proteomic characterization of several physique fluids.
A detailed molecular characterization from the synovial fluid could identify proteins connected with pathogenesis, which can be created as markers for evaluation in the disease in early stages MDV3100 clinical trial and its progression. Yamagiwa et al. demonstrated a 5 fold raise in the expression of 18 protein spots which includes haptoglobin among diverse synovial fluid samples from OA sufferers applying 2 DE platform. In yet another study, 135 proteins were identified from synovial fluid and 18 of them had been shown to become differentially expressed in OA sufferers. Pro teins identified to become elevated in OA included alpha 1 mi croglobulin, apolipoprotein E, complement component 3, haptoglobin, orosomucoid 1 and group particular compo nent. A approach of en dogenous profiling of peptides from OA synovial fluid that resulted in identification of 40 proteins was described by Kamphorst et al.
in 2007. Within a current study, abnor mally high levels of complement elements have been shown in OA synovial fluid. Sohn et al. identified 108 pro teins from OA synovial fluid and located that only selelck kinase inhibitor 36% of them were recognized to be in the plasma serum. Sixty six proteins, involved in acute phase response, comple ment and coagulation pathways have been reported to be differ entially expressed involving healthful and OA synovial fluid in a recent study by Ritter et al. A summary of earlier proteomic studies on OA synovial fluid is pro vided in Table 1. Most of these investigations had been carried out making use of low resolution mass spectrometers and with minimal fractionation of the samples, which limited the depth of coverage.
Within this study, we carried out a compre hensive cataloging of proteins from OA synovial fluid by like several fractionation approaches followed by higher resolution mass spectrometry analysis. Benefits and discussion Identification of proteins from OA synovial fluid Synovial fluid from five OA sufferers was pooled and the abundant proteins had been depleted using Human MARS 6 column. The resulting sample was then subjected to mul tiple fractionation techniques SDS Web page at the protein level and SCX and OFFGEL in the peptide level to minimize the complexity on the sample.

Background Hemophilia B is definitely the X linked monogenetic disorder triggered by the loss of functional coagulation issue IX, resulting within a deficiency inside the capacity of blood to clot. Moreover to enhanced propensity for bleeding following trauma or injury, spontaneous bleeds can take place in capillaries, specifically inside the joints, resulting in tissue harm more than time. Bleeds into essential closed spaces can be life threatening. At present, hemophilia B is treated by intravenous administration of F. IX concentrate, either plasma derived or recombinant, to be able to restore hemostasis. Because of the quick half life with the protein in circulation, frequent injections are required to supply prophylaxis or to treat sufferers with severe disease on demand.
Gene therapy represents an attrac tive option to protein replacement therapy, since it would involve a single injection to supply long-term in trinsic production of F. IX. Among potential gene therapies for hemophilia B, the usage of adeno associated virus as a gene delivery vector has shown essentially the most success to date. AAV can be a dependovirus, selleck EPZ005687 a parvovirus that may be unable to replicate in the absence of a helper virus. For use as a gene therapy vector, all viral genes are removed, leaving only the inverted terminal repeats expected for packaging about the transgenic construct. The several serotypes of AAV have different tropisms, which allow for gene transfer to several target tissues. For in stance, AAV1 can effectively transduce skeletal muscle, although AAV8 has powerful tropism for liver tissue. Pre clinical studies in animals established that the danger of immune responses to F.
IX is substantially impacted by the route of vector administration and by the underlying genetic defect. F9 null mutations are probably related with powerful immune response, even though mutations preserving Crizotinib some level of endogenous, albeit non functional F. IX expression, lessen the risk for immune responses. Current clinical trials are based on liver directed gene transfer. Hepatocytes are the typical web-site of F. IX syn thesis. In addition, high levels of antigen expression in hepatocytes promote induction of regulatory T cells, resulting in immune tolerance induction to the trans gene product. This approach is even in a position to reverse an ongoing antibody response against F. IX. Sustained expression of F.
IX by hepatic gene transfer has now been demonstrated in hemophilia B sufferers, following suc cesses in significant animals model, such as non human primates and hemophilia B dogs. AAV vectors traditionally contain a single stranded DNA genome using a packaging limit of ap proximately 5 kb. By modifying one of the inverted terminal repeats, it is probable to force the virus to pac kage a self complementary double stranded DNA ge nome, thereby bypassing the want to for second strand synthesis, one of the price limiting actions in AAV transduction.

There was no mention concerning MMP expression right after TLR blockade, and it remains unclear whether or not TLR is involved in MMP expression within a additional direct way. Our preliminary benefits have shown that S. aureus culture supernatant and complete cell lysate induce the mRNA expression of quite a few members on the TLR family members, which includes TLR 2. To elucidate the MMP induction by S. aureus, we turned to two effectively characterized mutant strains of S. aureus lacking Sar A and Agr. Agr and Sar would be the two finest characterized loci accountable for modulating the expression of S. aureus viru lence components. S. aureus strains lacking either locus have already been shown to result in attenuation of S. aureus in various models of staphylococcal diseases. Current investiga tions by Blevins and colleagues have also shown that mutation of Sar A and or Agr triggered lowered capacity to induce both SA and osteomyelitis.
The precise mechanisms of lowered effectiveness of Sar Agr mutants to bring about SA or osteomyelitis aren’t identified. Research by Nilsson selleck p53 inhibitor and col leagues showed that mice inoculated with all the Sar A sta phylococcal strain exhibited a a lot more pronounced T and B lymphocyte activation and larger levels of serum IL 6 and IFN, compared having a Sar A mutant, and infection with Sar A staphylococci induced pronounced fat reduction also. These research recommended that Sar A locus may possibly control molecules that happen to be significant virulence determinants inside the induction and progression of SA. We therefore tested the MMP 1, three, and 13 expression patterns in response to Sar, Agr, or Sar Agr mutants in human dermal fibroblasts.
The three MMPs have been selected as a result of their recognized involvement in a variety of models of arthritis and their respective degrading actions on collagen variety I, II, and III and proteoglycan, selleckchem that are crucial constit uents of connective tissues and cartilage in the joints. Our final results didn’t show any considerable differences in MMP 1 and MMP three mRNA levels, and 13 mRNA levels have been minimal and could not be quantified with affordable accuracy in dermal fibroblasts upon exposure to culture supernatants or cell lysates obtained in the mutants and isogenic parent strain. Nevertheless, interestingly, the expression of TIMPs was notably enhanced in fibroblasts treated with Sar Agr mutants compared with isogenic parent strain. This could mean that the effective biologically active MMPs are less abundant in cells treated with the Sar Agr mutants com pared with cells treated with isogenic parent strain.
It will likely be essential to estimate the levels of biologically active MMPs to identify the net effect of Sar Agr mutants on MMP expression. Temporal estimation of biologically active MMPs inside the joints immediately after infection with isogenic parent and mutants will help to clarify the issue of MMPs as a factor inside the observed differences in severity of ailments brought on by wild kind and mutant strains.

We previously reported that ApoE3 induces bAPP expression but ApoE4 does not, confirming the findings of Ezra et al. In this regard, elevations of ApoE by the method of neuroinflammation, or other stressors, would reflect a requisite part for the lipopro tein in enhancing the useful roles of bAPP and or other acute phase response proteins. Thus, it could be the inability of ApoE4 to participate in this chain of salutary events that makes it detrimental. We’ve pre viously shown that the enhance in ApoE brain levels that happens with aging continues to occur in AD, with a large fraction being deposited in plaques. This increase in ApoE levels is distinguishable from alterations in bAPP, which rises with age but declines markedly in AD.
This illness linked severance in the coor dinate regulation of ApoE and bAPP additional strengthens the correlation of brain wellness together with the coregulation of these two proteins, to wit, with ApoE expression itself, provided that the ApoE will not be ApoE4. Multi MLN9708 lineage kinase pathways could be invoked inside the regulation of ApoE expression, and may themselves be invoked by ApoE, suggesting a feedback loop in between MLK pathways and ApoE expression in neu rons. Our findings of involvement of multiple MLKs ERK, p38 MAPK, and JNK in expression of ApoE in neurons exposed to IL 1b, Ab, or sAPP, collectively with prior reports of ERK pathway invocation of ApoE expression and vice versa, are consistent together with the exis tence of a complex feedback method that may be impor tant in acute phase responses to neuronal injury as well as prospective exacerbation of neurodegenerative events.
Our obtaining that glutamate regulates supplier MG-132 ApoE expression via ERK and JNK, but not by p38 MAPK pathways may possibly be indicative of a correlation amongst glutamatergic induction of ApoE and neuronal survival. Excitotoxic effects of glutamate are largely dependent upon activa tion of extrasynaptic NMDA receptors, p38 MAPK, and the inhibition of ERK signaling, synaptic receptors, alternatively, appear to activate ERK and market survival. In conclusion, the induction of neuronal ApoE by either neuroinflammatory or excitotoxic agents or neu rotoxins, acting by way of MLK pathways suggests that alterations in these signaling pathways, with each other with other neuropathological entities in AD brain, may have consequences for ApoE expression.
Differences within this expression might be crucial, taking into consideration the role of APOE genotype in AD danger. The response of ApoE to IL 1b we show right here in rodent brain suggests that elevation of IL 1 results in the increases in ApoE that we and other folks have observed inside the AD brain. This may have added significance with regard towards the self propagating nature of IL 1 driven cascades, specially when such cascades are instigated inside the context of an ?four allele of APOE.

Several research have previously reported the OA effects around the cell metabolism. Cable et al. observed that OA impacted the heme metabolism of human hepatic cell lines. Also, Shisheva and Shechter showed that OA mimicked some of insulin bioef fects stimulating the glucose and lipid metabolism in rat adipocytes, and Tanti et al. found that glycolysis was stimulated and glucose transport was enhanced right after OA remedy in mouse skeletal muscle. Far more recently, an additional study showed that OA depressed the metabolic price of rat hepatocytes and changed glucose uptake in these cells. Related to electron transport chain, OA was previously found to induce alterations in mitochondrial membrane potential and elevated oxidative anxiety in the rat brain after intracerebroventri cular injection, and in distinctive cell sorts in vitro.
The altered expression levels in genes related to cell metabolism and electron transport chain identified within this study could help to clarify the effects described in all these works. Apart from, 8% of your genes altered right after the 3 h OA treatment have been related to cellular transport processes. OA was previously hop over to this website discovered to interfere inside the secretion of newly synthesized proteins and exocytosis in rats. both effects may be associated with the expres sion alterations identified inside the present study. When cells have been treated with OA for 24 h, the obtained genes were also categorized into diverse groups which includes translation, signal transduction, elec tron transport chain and redox homeostasis, metabo lism, cell cycle and apoptosis, transcription and nuclear precise proteins, transport, and cytoskeleton and cell adhesion.
Similar for the 3 h OA remedy, an important quantity of these genes are involved in metabolism like electron transport chain, but in addition a terrific percentage of genes related to selleck chemicals OC000459 translation had been observed. The expression alterations discovered within the genes involved in processes of translation and transcription may well be related to the previously reported OA induced inhibition of protein synthesis. Among the genes altered following the 48 h OA remedy, most were associated with signal transduction, translation, cell cycle and apoptosis, electron transport chain and redox homeostasis, metabolism, cytoskeleton and cell adhesion, transcription and nuclear particular proteins, and transport. Fewer genes related to metabo lism and transcription had been discovered altered at 48 h, but similarly to 24 h a crucial percentage of altered genes are involved in cell cycle and apoptosis. In another preceding study, the gene expression alterations in mouse BALBc3T3 cells soon after various OA treatment instances were evaluated by microarray analysis, plus a total of 177 differentially expressed genes have been identified.