Three drops cell suspension were placed in a single well of a sta

Three drops cell suspension were placed in a single well of a standard 12 well culture plate. The cells were allowed to adhere for two hours at 37 C, then 1 ml maintenance medium was added to each well. Geneticin or puromy cin pressure was maintained during chondrogenesis. Micro masses were cultured in the maintenance med ium sellectchem containing an ITS premix and 5 ug ml human transferrin for two weeks. The mineralization phase was induced using a MEM medium containing 5% fetal bovine serum, ITS premix, 5 ug ml human transferrin and 7 mM beta glycerolphosphate from Day 14 until Day 21. Each condition was performed in tripli cate. Total RNA from micro masses was isolated after 7, 14 or 21 days in culture using the Nucleospin RNA II kit.

Protein extraction of the micro masses stably overex pressing FRZB or controls after seven days was per formed using cell extraction buffer supplemented Inhibitors,Modulators,Libraries with 1 mM phenylmethanesulfonyl and 5% protease inhibitor cocktail, followed by quantification using the Pierce BCA Protein Assay kit. Some ATDC5 micro masses were fixed in 95% ice cold methanol for staining. For Picrosirius Red, Inhibitors,Modulators,Libraries micro masses were stained for one hour in Picrosirius Red in a saturated aqu eous solution of picric acid washed three times with 0. 5% acetic acid in water and air dried. For Safranin O, micro masses were stained for one hour in Safranin O washed three times with water and air dried. Quantifica tion of the staining was performed by dissolving the micro masses with 1 M NaOH or 6M Guanidine HCl and by measuring the absorbance at 540 and 512 nm respectively with the Infinite M200.

cDNA synthesis and Quantitative Real Time PCR Complementary Inhibitors,Modulators,Libraries DNA was synthesised from 1 ug of RNA isolated from tibia articular cartilage and For TaqMan assays analysis was performed using the PerfeCTa qPCR FastMix UNG using the following conditions, 1 minute at 95 C, 40 cycles of 3 seconds of denaturation at 95 C, followed by 20 seconds of annealing extension at 60 C. All experiments were performed in duplicate. For SYBRgreen, quantitative analysis was performed as follows, 10 minutes at 95 C, 40 cycles of 15 seconds of denaturation at 95 C, fol Inhibitors,Modulators,Libraries lowed by 60 seconds of annealing extension at 60 C. Melting curve analysis was performed to ensure amplifi cation of a specific product. The Corbett Rotor Gene 6000 was used for both systems.

Results are expressed using the comparative threshold method and were normalised to housekeeping gene Hprt. Mouse rib chondrocyte isolation and proliferation analysis Rib and sternum chondrocytes were isolated from three six week old wild type and three Inhibitors,Modulators,Libraries Frzb mice, as described with minor modifications. The sternum selleck chemicals was longitudinally cut, followed by complete removal of the ventral part of the ribcage. The ribcage was washed three times in Dulbeccos phosphate buffered saline with 1% AB.

AP 2a has previously been demonstrated to interact with the SUMO

AP 2a has previously been demonstrated to interact with the SUMO E2 conjugating enzyme, UBC9, in a yeast two hybrid screen, but to confirm that AP 2a can be sumoylated in vivo, expression constructs for UBC9 and SUMO 1 or SUMO 2 were cotransfected in HepG2 cells together with the different AP 2a isoforms. This resulted in the appearance, for isoform 1a only, of a slower considering migrating band at approximately 75 kDa, compatible with a mono sumoylated form of AP 2a. Further more, mutation of lysine 10 led to a very pronounced reduction in intensity of this sumoylated band, thus confirming that lysine 10 is the predominant sumoyla tion site Inhibitors,Modulators,Libraries Inhibitors,Modulators,Libraries in isoform 1a, and explaining why the other isoforms were not significantly modified.

To confirm that the inhibitory activity of isoform 1a is dependent on sumoylation, HepG2 cells Inhibitors,Modulators,Libraries were cotrans fected with the 3xAP2 Bluc reporter and increasing doses of SUMO 1 or SUMO 2. The transactivation activ ity was reduced with SUMO co transfection, particularly by SUMO 2, while the transactivation activity of the K10R mutant or AP 2a 1c was not altered significantly, suggesting that the most important sumoylation site is indeed lysine 10 of isoform 1a. To confirm that the inhibitory effect exerted Inhibitors,Modulators,Libraries by isoform 1a on the cyclin D3 reporter is due to sumoylation, we co trans fected with either wild type UBC9 or its sumoylation defective mutant, C93S. While transfection of wt UBC9 did not result in further inhibition of cyclin D3 reporter activity, cotransfection of UBC9 C93S partially reverted the inhibitory activity of isoform 1a.

In addition, a clear reduction in the level of sumoylated Inhibitors,Modulators,Libraries http://www.selleckchem.com/products/brefeldin-a.html AP 2a was observed by western blot when the 1a isoform and UBC9 C93S were co expressed in the same propor tion in HepG2 cells. TFAP2A isoform 1a is a weaker transactivator of the ERBB2 promoter The transactivation activity of the different AP 2a iso forms was further tested on the ERBB2 promoter, which is known to possess two functional AP 2 binding sites, mapping at 210 and 500 upstream of the transcription start site, which are required for positive regulation of ERBB2 by AP 2a in breast tumour lines. A repor ter carrying ERBB2 promoter sequences was co transfected at different ratios with the AP 2a isoform expression constructs together with Cited2 p300. Isoform 1a significantly induced reporter activity at a 1,2 and 1,4 reporter,AP 2a ratio. In contrast, the transactivation activity of isoforms 1b and 1c was already significant at a ratio of 1,0. 5, and further increased at higher ratios, reaching a plateau at 1,4. Iso form 1d was the most potent transactivator and its activ ity increased at higher ratios in an almost linear manner.

Culture of primary HACs HACs from OA cartilage portions with less

Culture of primary HACs HACs from OA cartilage portions with less than 50% of thickness loss were selleck screening library released by enzymatic digestion, as previously described. Isolated chondrocytes were plated in 100 mm diameter Inhibitors,Modulators,Libraries dishes and cultured to 70% confluence in Dulbecco Modified Eagle Medium containing 10% fetal bovine serum, 100 IU ml peni cillin, and 100 ug ml streptomycin at 37 C in a humidified 5% CO2 atmosphere. After HACs were transferred to six well plates, they were stimulated for 4 hours with IL 1B in serum free media. The SOCS1 overexpressing HACs were cultured in pellets 24 hours before the stimulation with IL 1B. PCR products were digested with BamH1 and EcoRI and cloned into the pBABE viral vectors. To produce retrovirus, the pBABE SOCS1 viral vectors were trans fected into a Platinum A retroviral packing cell line.

Su pernatants were collected Inhibitors,Modulators,Libraries 72 hours after transfection. To infect SW1353 cells, viral supernatant was mixed with fresh medium with 8 ug Inhibitors,Modulators,Libraries ml of polybrene at 1,1 ratio, and the mixture was applied to freshly seeded cells. To deliver SOCS1 or control shRNA into the SW1353 cells, SOCS1 shRNA or copGFP lentiviral particles were mixed with fresh medium and 5 ug ml of polybrene, and the mixture was applied to freshly seeded cells. Stable overexpressing or knockdown cell lines were selected with puromycin. To establish SOCS1 overexpressing HACs, pShuttle2 SOCS1 or empty vector was electro transfected by using a Gene Pulser Xcell System under the condition of 50 V and 2 ms pulse.

Measurement of MMPs and TIMP 1 in culture supernatants Nontransfected and transfected SW1353 cells were plated onto 48 well plates for 24 hours and then pretreated with Inhibitors,Modulators,Libraries serum free media for 2 hours. The cells were stimulated with IL 1B for 24 hours. The concentrations of MMP 1, 3, and ?13 and TIMP 1 in the conditioned media were measured with commercial ELISA kits according to the manufac turers instructions. Western blotting and immunoprecipitation To prepare the total cell lysates, SW1353 cells were washed twice with ice cold PBS, harvested, and lysed in IP buffer, 150 mM NaCl, 1% Triton X 100, 25 mM B glycerophosphate, phosphatase inhibitor cocktail, and protease inhibitor cocktail for 60 minutes on ice. For Inhibitors,Modulators,Libraries immunoprecipitation, TAK1 antibody was added to the whole cell extracts and incubated on a rotator overnight at 4 C.

Then the protein G agarose beads were further incu bated for 3 hours at selleckchem MEK162 4 C. The mixtures were centri fuged at 2,095 g for 3 minutes. The precipitates were washed 3 times by using pre cold IP buffer, and the beads were resuspended in 2�� SDS sample buffer. The immunoprecipitates or the whole cell lysates were separated on 10% denaturing polyacrylamide gels and transferred to polyvinylidene difluoride membranes. The membranes were probed with appropriate primary anti bodies and IgG horseradish peroxidase conjugated anti bodies. Signals were developed by using an enhanced chemiluminescence system.

Conclusions In summary, our

Conclusions In summary, our selleck products findings suggest that long term endo crine Inhibitors,Modulators,Libraries therapy facilitates translocation of GPR30 to cell membranes, resulting in inappropriate activation of the EGFR signaling pathway. Meanwhile, GPR30 attenuates the inhibitory effect of cAMP on MAP ki nases. Combination treatment with the GPR30 specific antagonist G15 plus Tam induces both cytocidal action in vitro and antitumor progression in vivo. Thus, GPR30 might be a useful target in developing better treatments for TAM R breast cancer patients. In the last decade, genomic studies have identified five breast cancer intrinsic subtypes, basal like and clau din low . In a recent study, an integrated analysis of copy number and gene expression split the intrinsic subtypes revealing novel subgroups with distinct clinical outcome, including a high risk ERa positive subgroup and a subset of ERa positive and ERa negative cases with a favorable outcome.

According to this analysis, the majority of the basal like tumors formed a high genomic instability subgroup Inhibitors,Modulators,Libraries with relatively good long term outcomes. Basal like pheno types represent tumors that express markers that are characteristic of the myoepithelium of the normal mam mary gland, such as epidermal growth factor receptor, p63 and the basal cytokeratins CK14, CK5 6 and CK17. They show partial overlap with the tri ple negative breast cancers that are characterized by a lack of HER2 gene amplification and estrogen and pro gesterone receptor expression. Inhibitors,Modulators,Libraries Approximately 75% of triple negative breast cancers are classified as basal like tumors on the basis of their overall gene expression pro file.

The basal like phenotype represents a more homoge neous group of cancers than the group of cancers defined by triple negativity. Basal like tumors are often Inhibitors,Modulators,Libraries resis tant to chemotherapy and develop distant metastases in characteristic tissues, such as lung and brain. Recent studies have suggested a correlation between the basal phenotypes and epithelial to mesenchymal transition. EMT has been reported to promote invasion during the progression of breast carcinomas and it is considered as an essential early step in tumor metastasis. EMT is characterized by loss of cellular adhesion, which is mediated by down regulation of adhesion molecules, such as CD44 and E cadherin. The expression of E cadherin is regulated Inhibitors,Modulators,Libraries by a number of transcriptional repressors, which include SNAIL, SLUG, SIP 1, EF1 and TWIST.

The family of micro RNAs 200 and the miR 205A regulate the expression of the transcriptional repressors of E cadherin ZEB 1 and ZEB 2 and, consequently, the levels of E cadherin in breast cancer cells and tissues. A decrease in the expres sion of these microRNAs has been observed never in cells that have undergone EMT and in mesenchymal regions of metaplastic breast cancer lacking E cadherin expression.

In the presence of IGF 1, glucose decreased the secretion proges

In the presence of IGF 1, glucose decreased the secretion proges terone and oestradiol by a factor of selleck chemical Belinostat about three. Similar results were obtained with a lower dose of glucose. Thus, a high glucose concentration decreased both basal and FSH or IGF 1 stimulated pro gesterone and oestradiol production in rat granulosa cells. We then investigated whether this inhibitory effect of glu cose on the production of both progesterone and oestra diol resulted from the production of smaller amounts of the three key enzymes in steroidogenesis and or of StAR, a major cholesterol carrier. Glucose treatment in the presence of FSH decreased production of 3 HSD and Inhibitors,Modulators,Libraries p450scc by a factor of about seven, halved the production of StAR and reduced by three fold the production of p450 aromatase, relative to the values in the presence of FSH without glucose.

In the presence of IGF 1, glucose decreased the amounts of the three Inhibitors,Modulators,Libraries key enzymes in steroidogenesis and StAR by a factor of three relative to IGF 1 treatment without glucose. Similar results were obtained with a lower glucose concentration. In the basal state, glucose treatment only halved the production of 3 HSD, StAR and p450 aromatase but did not affect the amount of p450scc protein. Thus, the decrease in FSH or IGF 1 induced progesterone and oestradiol secretions in response to glucose treatment appears to be caused by a reduction in the amounts of the 3 HSD, p450scc, StAR and p450 aromatase proteins. The inhibition of basal progesterone and oestradiol secretions in response to glucose could be the result of a reduction in production of the 3 HSD, StAR and p450 aromatase pro teins.

Effects Inhibitors,Modulators,Libraries of glucose on granulosa cell proliferation and viability We investigated whether the dose of glucose used affected the number of granulosa cells in culture, either by induction of mitosis or by altering the cell viabil ity. thymidine incorporation by granulosa cells treated with 10 g l glucose was determined after 24 h of culture in the presence or in the absence of FSH or IGF 1. As expected, FSH and IGF 1 treatment significantly increased thymidine incorporation. However, glucose treatment did not affect cell proliferation or cell number. Glucose had no effect on the cell viability in the absence or in the presence of FSH and IGF 1 as assessed by staining with Trypan blue.

Effects of glucose treatment on the MAPK ERK1 2 and AMPK phosphorylation and on the adiponectin receptor expression in rat granulosa cells We examined whether the inhibitory effect of Inhibitors,Modulators,Libraries glucose on progesterone and oestradiol Inhibitors,Modulators,Libraries production was associated with a variation in the phosphorylation of MAPK ERK1 2 and AMPK. These kinases have been implicated in the reg ulation of steroidogenesis. http://www.selleckchem.com/products/Gemcitabine-Hydrochloride(Gemzar).html We analysed the pat tern of MAPK ERK1 2 phosphorylation in cells incubated with 10 g l glucose for various times, or with 10 g l glucose for 48 h in the pres ence or absence of FSH or IGF 1.

Mitochondrial protein was collected after centrifuging at 15,000

Mitochondrial protein was collected after centrifuging at 15,000 rpm for 30 min at 4 C, aliquot and stored at ?70 C. Western blot inhibitor Dovitinib analysis of growth regulatory proteins and apoptosis proteins Cells were treated with ZD6474 and or UV B and then the cells were scraped and lysed in Nonidet P 40 lysis buf fer containing 1 mM sodium vanadate, 1 mM phenylmethylsulfonyl Inhibitors,Modulators,Libraries fluoride, and protease cocktail in hibitor for obtaining total cell extracts. Equal amount of cell extracts were separated on a 10% sodium dodecyl sulfate polyacrylamide electrophoretic gel and transferred to nitrocellulose membranes, which were blocked with 2% BSA and probed with the appropriate antibodies and secondary antibodies. Membranes were then developed using enhanced chemiluminescence or al kaline phosphatase based colorimetric methods.

Caspase 3 and caspase 7 activity assays Caspase 3 and Inhibitors,Modulators,Libraries Inhibitors,Modulators,Libraries caspase 7 activity was determined by meas uring the absorbance at 405 nm after cleavage of synthetic substrate acetyl Asp Glu Val Asp p nitroanilide as described previously with some modifications. Cells were treated with ZD6474 and or UV B radiation for 48 h, and lysed with buffer, followed by centrifugation at 20,000 g for 15 min at 4 C. The lysates were incubated in 200 uM solu tion of in a reaction buffer at 37 C. The reaction was monitored for 1 3 h, and the ab sorbance was recorded at 405 nm. If the signal was low, the reaction can be monitored for 12 24 h. The formation of pNA was calculated as the difference in the absorbance at 405 nm unit time per unit volume of sample.

The relative levels of pNA formation were normalized against the protein concentration of each extract to obtain specific Inhibitors,Modulators,Libraries activity. In vitro wounding assay To test the invasive behavior of treated cells, 1 105 cells were plated in 6 well tissue culture plates and grown for 24 h to obtain a confluent monolayer and migration was studied by in vitro wounding assay with slight modifications. The monolayer was scraped in a straight line to create a wound with a p200 pipette tip. The debris were re moved and the edge of the wound was made smooth by washing the cells once with 1 ml of the growth medium and then replaced with 3 ml of complete Inhibitors,Modulators,Libraries media along with ZD6474 and or UV B. Cells were observed 48 h post treatment. Cells invading the wound line were observed under an inverted phase contrast microscope.

The dis tances between one sides of the scratch with another inhibitor price were measured after the indicated time intervals using the Leica Qwin software. The distance of each wound clo sure was the measure of wound healing. P values of wound size were calculated using un paired t test between the same treatment group, prior and post treatment. Each experiment was performed three times with triplicate samples. Scanning electron microscopy Cells were grown in cover slip at a density of 10,000 cells per cover slip.

Likewise, knock out of KRASG13D in DLD 1 cells signifi cantly rev

Likewise, knock out of KRASG13D in DLD 1 cells signifi cantly reverted the migration ability of DLD 1 cells. BRAFV600E enhances the ability of Caco 2 cells to migrate and invade in vitro www.selleckchem.com/products/Lenalidomide.html through RhoA activation Overexpression of BRAFV600E in Caco 2 cells had a pro found effect on the RAS effector Inhibitors,Modulators,Libraries protein RhoA. In Caco BR cells activation of RhoA is increased as well as phosphorylation of its down stream target Cofilin, a protein that is related to stress fibre formation. These findings are closely related to the observation regarding increased stress fibre formation indicated by phalloidin staining in Caco BR13 cells. Notably, an extra band of lower molecular weight is detected for RhoA in Caco BR and DLD 1 cells, which potentially represents the main active GTPase form.

A variant of lower molecular weight for RhoA protein has previously been reported both in colon and breast tissues. However, RT PCR analysis and treatment with the proteasome inhibitor MG 132, both in Caco BR and DLD 1 cells, suggested no association of this faster migrating RhoA band with alternative Inhibitors,Modulators,Libraries splicing or proteasomal Inhibitors,Modulators,Libraries degrada tion. These data suggested that the additional band potentially represents a post transla tional modification of RhoA protein. To further explore the role of BRAFV600E in the activation of the RhoA pathway, transient transfection of the oncogene in Caco 2 cells was performed. Subsequent analysis of the migration and invasion properties showed Inhibitors,Modulators,Libraries that moderate RhoA activation induced a partial cell migration and cell invasion response.

Notably in the invasion assay cell phenotype became slightly altered and resembled that of the stable Caco BR clones, suggesting that a stable expression of BRAFV600E is required to achieve complete cell transformation and extensive RhoA activation. Regarding the importance of RhoA activation in the induced cell migration and invasion observed in Caco BR cells, siRNA against RhoA Inhibitors,Modulators,Libraries was performed leading to significant protein depletion in both Caco 2 and Caco BR13 cells. Depletion of RhoA substantially impaired both acquired properties with more profound effect in Caco BR13 cells, further illustrating its central role in the BRAFV600E oncogene induced transformation of colon adenocarcinoma cells. Moreover, following RhoA depletion in Caco 2 cells, the number and size of stress fibres were notably reduced as com pared to Caco BR cells, where no such alteration was observed.

In order to study further the impact of RhoA GTPase on cell migration, silencing of RhoA was performed in DLD 1 and HT29 cells. Considering that these cell lines bear mutation in KRASG13D and BRAFV600E respectively, RhoA depletion was also performed in selected 17-AAG Sigma clones where KRASG13D or BRAFV600E was knocked out or down regulated via shRNA respectively. This approach can implement the connection between each oncogene and the small GTPase.

A total of 38 rDNA units organized in transcriptional units, incl

A total of 38 rDNA units organized in transcriptional units, includ ing a small subunit rRNA gene, a 5. 8S rRNA gene, and a large subunit rRNA gene in a 5 3 orientation, have been detected in the genome. The sizes of the small subunit, 17-AAG msds the large subunit and the 5. 8S rRNA gene are 1. 8 kb, 2. 45 kb and 0. 44 kb, respectively. Some units are tan demly duplicated, up to four copies on scaffold 18, and some may also be localized in subtelomeric regions, as revealed by a co mapping of telomeric sequences and rDNA subunits at scaffold 6 and 9 extremities. These two scaffolds could correspond to entire chromosomes. Inhibitors,Modulators,Libraries Due to the sequencing method, some units are incomplete. The alignment of 20 complete small subunit rRNA genes shows polymorph ism between copies, which is also the case for 29 large subunit rRNA gene copies.

Inhibitors,Modulators,Libraries The number of genes in Blastocystis is Inhibitors,Modulators,Libraries reduced in comparison with other stramenopiles. Surprisingly, a large por tion of genes were probably duplicated since 404 clusters of paralogous protein coding genes were identified, Inhibitors,Modulators,Libraries con taining 1,141 genes, that is, 19% of Blastocystis genes. Excluding the large multigenic families, most of the dupli cated genes are present in only two copies. As described in other organisms, the duplicated genes are more conserved than single copy genes in Blastocystis sp. Indeed, they have more orthologs. see Materials and methods and display higher similarities with their ortho logs. They also tend to display higher expression levels than single copy genes. We investigated whether these gene duplications could have arisen from a whole genome duplication or smaller scale segmental duplications.

WGD, the duplica tion Inhibitors,Modulators,Libraries of the entire genome by polyploidization, has been shown to have played a key role in the evolutionary history of several animal and plant lineages. Segmental duplications occur continually by several mechanisms that can duplicate parts of genes, entire genes, or several adjacent genes. These mechanisms include unequal crossing over, or gene conversion, and tandem duplication. We were able to identify 320 blocks of duplicated genes, that is, paralogous seg ments of several adjacent genes, some of which are very large, suggesting a WGD. These blocks cover about 39% of the genome representing 38% of the unrepeated fraction of the gen ome. As shown in Figure 1, each scaffold is a mosaic of blocks of homology with several other scaffolds scaf folds cannot be grouped by pairs as would be expected from a recent WGD. Additionally, some segments are present in more http://www.selleckchem.com/products/Perifosine.html than two copies in the genome, suggesting that segmental duplications are likely to have played a role in the current duplication pattern.

Oestrogen deprivation resistant MCF7 X cells differ in that they

Oestrogen deprivation resistant MCF7 X cells differ in that they do not have up regulation of HER2. and indeed such growth factor receptors selleck products only rarely increase in clinical endocrine relapse samples. It is thus probable that some dependence on the PI3K Akt pathway in MCF7 X cells coupled with only low HER2 activity cumulatively results in their prominent growth insensitivity to RAD001. Rapalogue insensitive acquired endocrine resistant cells retain sensitivity to mTOR kinase inhibition Given their substantial RAD001 growth insensitivity, TamR and MCF7 X cells could comprise useful models to under stand everolimus resistance in tamoxifen or oestrogen deprivation resistant patients and to determine improved treatments. In Inhibitors,Modulators,Libraries this regard, our findings suggest that inhib ition of both TORC2 Akt and TORC1 may be critical.

We have shown that growth of cancer cells unresponsive to an allosteric mTOR inhibitor can still be sensitive to mTOR kinase blockade, with AZD8055 substantially inhibiting samples and our additional results here are encour aging in that they show AZD8055 is also highly growth inhibitory Inhibitors,Modulators,Libraries in a further ER acquired endocrine resistant model, T47D tamR. Our TamR and MCF7 X signalling studies indicate that the difference in the mechanism of action of these drugs enables AZD8055 to target mTORC2 signalling in addition to mTORC1. thus, basal p Aktser 473 is rapidly and substantially inhibited by AZD8055 in both models but is unaffected by RAD001. Furthermore, inhibition of a negative feedback loop downstream of mTORC1 can result in induction of Akt activity in patients treated with everolimus.

This event may limit the effectiveness of mTORC1 targeting therapy since it can be associated with shortened time to progression in pa tients. However, in both Inhibitors,Modulators,Libraries TamR and MCF7 X cells, basal Akt ser473 inhibition by AZD8055 was sus tained with no up regulation over 24 hours treatment. A similar response to AZD8055 has been reported in fur ther breast cancer cell lines, while clinically a close analogue of AZD8055, AZD2014, also inhibits Inhibitors,Modulators,Libraries pAKT, pS6 and p4EBP 1 in some tumours. AZD8055 has previously been reported to induce cell death and autophagy in lung and leukaemia cancer cells, contrasting rapalogues that are often poor inducers of cell death. In the present study, some cell death was induced in TamR but not in MCF7 X cells by AZD8055. Since bcl 2 mRNA was detected basally in MCF7 X, but Inhibitors,Modulators,Libraries not in TamR cells, this anti apoptotic factor may contribute towards the somewhat reduced AZD8055 sensitivity in MCF7 X versus TamR cells. Interest ingly, rapamycin resistant tumours have previously been reported to express high levels of bcl 2.