epithelial - mesenchymal transition ( emt ) plays an important role in cancer tumorigenesis . during the emt ,
epithelial cells change their polarized epithelial phenotype to a spindle - shaped , myofibroblast - like phenotype with high motility , and cancer cells increased cell migration and invasion , which are crucial to cancer prognosis .
the partial loss of e - cadherin is generally accepted as a hallmark of the emt , which reduces cell - cell adhesion and destabilizes the epithelial architecture .
moreover , vimentin bestows a motile phenotype to cancer cells through changes in cellular architecture and cell - matrix interactions .
further , many transcription factors , such as snail , act as repressors of e - cadherin   and have been linked to the induction of the emt under different cellular contexts .
signal transducer and activator of transcription 3 ( stat3 ) is also involved in emt by regulating the transcriptional regulators of e - cadherin .
large studies indicated that the emt was induced by transforming growth factor 1 ( tgf-1 ) in various cancer cells .
reported that tgf-1 was overexpressed in anaplastic thyroid cancer , and they found that knockdown of tgf-1 could inhibited cell proliferation and colony formation , and promoted apoptosis in anaplastic thyroid cells .
park et al . found that tgf-1 promotes cancer immune escape , and that it promoted cell proliferation , colony formation , and inhibited apoptosis .
tgf- is known to activate akt through pi3k , which in turn activates the mtor complex 1 ( mtorc1 ) .
the activation of mtorc1 affects cell growth , proliferation , and invasion by modulating protein synthesis through its downstream effector , eukaryotic translation initiation factor p70 s6 kinase .
recently , studies reported that sorafenib combined with a mammalian target of rapamycin ( mtor ) inhibitor was a more effective and tolerable treatment strategy for advanced hcc .
inhibition of the mtor pathway reduced migration and invasion ability , and knockdown of mtorc1 induced mesenchymal - epithelial transition .
recent studies showed that pkm2 ( pyruvate kinase m2 ) is needed to induce emt .
pkm2 is normally not expressed in adult tissues , but is reactivated in tumor tissues .
pkm2 controls the final rate - limiting step of glycolysis and is an alternatively spliced variant of the pkm gene .
moreover , pkm2 is a crucial glycolytic enzyme in the oncogenic mtor - induced warburg effect , in which hypoxia inducible factor-1 ( hif-1 ) and c - myc - hnrnp cascades are the transducers of mtor regulation of pkm2 .
in addition , a study reported that loss of snail inhibits cellular growth and metabolism through the mir-128-mediated p70s6k / pkm2 signaling pathway .
furthermore , atsushi et al . found that pkm2 knockdown failed to induce spindle - shaped morphological changes , and hindered e - cadherin reduction and vim increase compared with the control group .
we infer that the mtor / p70s6k / pkm2 pathway can regulate the emt state . in this study
, we hypothesized that the mtor / p70s6k / pkm2 pathway is involved in regulating tgf-1-induced emt .
we investigated how the mtor inhibitor regulated emt , and explored the mechanisms of tumor suppression .
our findings suggest that inhibition of mtor / p70s6k / pkm2 signaling promotes cervical tumor suppression .
cervical cancer cell lines ( hela , siha ) ( american type culture collection , manassas , va ) were grown in dmem , supplemented with 10% fetal bovine serum , 100 units / ml penicillin , and 100 g / ml streptomycin and maintained at 37c with 5% co2 .
cells were treated with or without 10 ng / ml tgf-1 ( peprotech , usa ) in a serum - free medium for 48 h. cells were incubated with 0 , 25 , 50 , and 100 nm of rapamycin ( cst , danvers , ma ) .
cell viability was assessed by use of a cell counting kit-8 ( cck-8 ) assay .
cells were plated in a 96-well plate at 310 cells / well in triplicates and treated with or without rapamycin and tgf-1 for 24 , 48 , and 72 h. ten l cck-8 was added to each well and incubated for 1 h at 37c .
cell viability was calculated by the following formula : cell viability ( % ) = ( od treatment  od blank)/(od control  odblank ) 100% .
cells were seeded in 6-well plates at 410 cells / well and then treated with 50 nm rapamycin with or without 10ng / ml tgf-1 for 24 h. apoptotic cells were detected by flow cytometry using an annexin v - fitc kit according to the kit instructions .
cells were seeded in 6-well plates , and were cultured with fresh serum - free medium containing tgf-1 with or without the indicated concentration of rapamycin for 24 h. cell monolayers were wounded by scratching with sterile plastic 200 l micropipette tips and photographed using phase - contrast microscopy .
the migration distance of each cell was measured after the photographs were converted to photoshop files .
scratch distance at 24 h)/scratch distance at 0 h. the cells were harvested by centrifugation and washed with pbs , and then they were lysed in ripa buffer containing protease inhibitors .
equal amounts of the protein lysates were electrophoretically separated on 10% sds - page gels and transferred to pvdf membranes .
the membranes were blocked with 5% nonfat milk and then incubated overnight at 4c with the primary antibodies : sanil2 , stat3 , phosphor - p70s6k , e - cadherin , vimentin , and -actin , which were purchased from cst .
after incubation with the secondary antibody for 1 h at room temperature , the protein bands were detected using the ecl detection system ( bd biosciences ) .
cervical cancer cell lines ( hela , siha ) ( american type culture collection , manassas , va ) were grown in dmem , supplemented with 10% fetal bovine serum , 100 units / ml penicillin , and 100 g / ml streptomycin and maintained at 37c with 5% co2 .
cells were treated with or without 10 ng / ml tgf-1 ( peprotech , usa ) in a serum - free medium for 48 h. cells were incubated with 0 , 25 , 50 , and 100 nm of rapamycin ( cst , danvers , ma ) .
cell viability was assessed by use of a cell counting kit-8 ( cck-8 ) assay .
cells were plated in a 96-well plate at 310 cells / well in triplicates and treated with or without rapamycin and tgf-1 for 24 , 48 , and 72 h. ten l cck-8 was added to each well and incubated for 1 h at 37c .
cell viability was calculated by the following formula : cell viability ( % ) = ( od treatment  od blank)/(od control  odblank ) 100% .
cells were seeded in 6-well plates at 410 cells / well and then treated with 50 nm rapamycin with or without 10ng / ml tgf-1 for 24 h. apoptotic cells were detected by flow cytometry using an annexin v - fitc kit according to the kit instructions .
cells were seeded in 6-well plates , and were cultured with fresh serum - free medium containing tgf-1 with or without the indicated concentration of rapamycin for 24 h. cell monolayers were wounded by scratching with sterile plastic 200 l micropipette tips and photographed using phase - contrast microscopy .
the migration distance of each cell was measured after the photographs were converted to photoshop files .
the cells were harvested by centrifugation and washed with pbs , and then they were lysed in ripa buffer containing protease inhibitors .
equal amounts of the protein lysates were electrophoretically separated on 10% sds - page gels and transferred to pvdf membranes .
the membranes were blocked with 5% nonfat milk and then incubated overnight at 4c with the primary antibodies : sanil2 , stat3 , phosphor - p70s6k , e - cadherin , vimentin , and -actin , which were purchased from cst .
after incubation with the secondary antibody for 1 h at room temperature , the protein bands were detected using the ecl detection system ( bd biosciences ) .
to detect whether tgf-1 induced emt , we used 10 ng / ml tgf-1 to stimulate hela and siha cells for 48 h. we observed that siha ( figure 1b ) and hela cells ( figure 1c ) cells became scattered , acquired a spindle - shaped morphology , and lost cell - cell contacts , which are characteristics of a mesenchymal - like morphology .
moreover , e - cadherin expression was abundant in the absence of tgf-1 ( pre - emt ) . when stimulated with 10 ng / ml tgf-1 , e
in contrast , when stimulated by tgf-1 , vimentin increased ( post - emt ) compared with the pre - emt state .
these changes in cell morphology and marker proteins indicate that emt was induced when siha and hela cells were stimulated with 10 ng / ml tgf-1 .
we evaluated the effect of inhibition of mtor pathway on suppressing proliferation of cervical carcinoma cells .
we used cell counting kit-8 ( cck-8 ) assays to determine the effects of rapamycin ( an mtor inhibitor ) with or without tgf-1 on cell proliferation .
cells were treated with 0 , 25 , 50 , and 100 nm rapamycin , and 50 nm rapamycin with or without 10 ng / ml tgf-1 . as shown in figure 2 , rapamycin inhibited the proliferation of in hela cells in a dose - dependent manner .
treatment with tgf-1 significantly increased the proliferation of both cell lines at 48 and 72 h , which was abolished by the addition of rapamycin ( figure 2a ) .
cells were treated with 50 nm rapamycin with or without 10 ng / ml tgf-1 .
tgf-1 significantly increased cell migration in hela cell lines at 24 h , which was abolished by the addition of rapamycin ( figure 3a ) .
similar data were obtained from the wound - healing assays of siha cells ( figure 3b ) . to evaluate the effect of inhibition of mtor for antagonizing the anti - apoptosis effect of tgf-1 on cervical carcinoma cells by annexin v - fitc and pi staining , we assessed the effect of rapamycin with or without tgf-1 on the apoptosis of hela ( figure 4a ) and siha ( figure 4b ) cells . when cells were treated with tgf-1 , the total number of apoptotic cells ( early apoptotic+apoptotic ) was significantly decreased compared to untreated cells in both cell lines . in cells treated with 50 nm rapamycin , the apoptosis rate significantly increased .
furthermore , the addition of rapamycin significantly abolished the tgf-1-induced anti - apoptosis effects in both cell lines . to determine whether the emt condition induces an increase in pkm2 , pkm2 expression was detected to compare levels pre- and post - emt state .
as showed in figure 1 , siha ( figure 1b ) and hela ( figure 1c ) cells changed morphology from epithelial to fibroblastic - like and spindle - shaped and lost cell - cell contacts , which are characteristics of a mesenchymal - like morphology .
we found that the level of e - cadherin expression was suppressed compared with the pre - emt state .
vimentin and snail family zinc finger 2 ( snai2 ) expressions were increased in the post - emt state ( figure 5 ) .
pkm2 expression was stimulated to increase in the post - emt condition ( figure 5 ) .
the data show that the induction of emt resulted in a decreased level of e - cadherin , and increased vim and pkm2 expression in hela ( figure 5a ) and siha cells ( figure 5b ) .
moreover , atsushiet et al . reported that they were able to knock down pkm2 under emt conditions , but pkm2 knockdown failed to induce spindle - shaped morphological changes .
meanwhile , pkm2 knockdown hindered e - cadherin loss and vim gain compared with the control .
we evaluated the effects of rapamycin ( an mtor inhibitor ) on pkm2 ( a critical glycolytic enzyme ) , and p70s6k ( s6k1 , a downstream effector of mtor ) . to investigate the inhibitory effect on mtor ,
we used 0 , 12.5 , 25 , 50 , and 100 nm rapamycin to treat hela ( figure 6a ) and siha ( figure 6b ) cells for 24 h. ribosomal p70 s6 kinase ( s6k1 ) is a main downstream mtor effector . as shown in figure 6 , rapamycin , a specific mtor inhibitor , inhibited the phosphorylation of p70s6k in a dose - dependent manner .
moreover , we investigated whether mtor / p70s6k signaling decreased pkm2 expression , which is a main downstream p70s6k effector .
we found that rapamycin treatment significantly decreased pkm2 in hela ( figure 6a ) and siha ( figure 6b ) cells .
based on these facts , inhibition of mtor appears to affect cervical cancer through the mtor / p70s6k / pkm2 pathway in cervical cancer cells . to determine whether rapamycin is involved in regulating emt in cervical cancer
tgf-1 significantly decreased the expression of e - cadherin and increased the expressions of vimentin , stat3 and snail2 in hela ( figure 7a ) and siha cells ( figure 8a ) .
in addition , at concentration of 50 nm rapamycin reversed tgf-1-induced emt - markers expression by repressing vimentin , stat3 and snail2 expressions and restoring e - cadherin expression in in hela ( figure 7a ) and siha cells ( figure 8a ) .
as shown in figures 7 and 8 , we found that tgf-1 significantly increased the phosphorylation of p70s6k , which is the main downstream signaling intermediate of mtor signaling .
simultaneously , tgf-1 significantly increased the expression of pkm2 in hela ( figure 7a ) and siha cells ( figure 8a ) , while rapamycin significantly decreased the expressions of p - p70s6k and pkm2 .
these results suggest that rapamycin reverses tgf-1-induced emt via the mtor / p70s6k / pkm2 pathway .
further , we examined the effect of rapamycin with or without tgf-1 on the morphology of hela and siha cell lines .
after stimulation with 10 ng / ml tgf-1 for 48 h , both hela ( figure 7b ) and siha cells ( figure 8b ) cells became scattered , acquired a spindle - shaped morphology , and lost cell - cell contact , which are characteristics of a mesenchymal - like morphology .
treatment with 50 nm rapamycin for 48 h abolished the tgf-1-induced morphological changes in siha and hela cell lines .
to detect whether tgf-1 induced emt , we used 10 ng / ml tgf-1 to stimulate hela and siha cells for 48 h. we observed that siha ( figure 1b ) and hela cells ( figure 1c ) cells became scattered , acquired a spindle - shaped morphology , and lost cell - cell contacts , which are characteristics of a mesenchymal - like morphology .
moreover , e - cadherin expression was abundant in the absence of tgf-1 ( pre - emt ) . when stimulated with 10 ng / ml tgf-1 , e
in contrast , when stimulated by tgf-1 , vimentin increased ( post - emt ) compared with the pre - emt state .
these changes in cell morphology and marker proteins indicate that emt was induced when siha and hela cells were stimulated with 10 ng / ml tgf-1 .
we evaluated the effect of inhibition of mtor pathway on suppressing proliferation of cervical carcinoma cells .
we used cell counting kit-8 ( cck-8 ) assays to determine the effects of rapamycin ( an mtor inhibitor ) with or without tgf-1 on cell proliferation .
cells were treated with 0 , 25 , 50 , and 100 nm rapamycin , and 50 nm rapamycin with or without 10 ng / ml tgf-1 . as shown in figure 2 , rapamycin inhibited the proliferation of in hela cells in a dose - dependent manner .
treatment with tgf-1 significantly increased the proliferation of both cell lines at 48 and 72 h , which was abolished by the addition of rapamycin ( figure 2a ) .
cells were treated with 50 nm rapamycin with or without 10 ng / ml tgf-1 .
tgf-1 significantly increased cell migration in hela cell lines at 24 h , which was abolished by the addition of rapamycin ( figure 3a ) .
similar data were obtained from the wound - healing assays of siha cells ( figure 3b ) . to evaluate the effect of inhibition of mtor for antagonizing the anti - apoptosis effect of tgf-1 on cervical carcinoma cells by annexin v - fitc and pi staining , we assessed the effect of rapamycin with or without tgf-1 on the apoptosis of hela ( figure 4a ) and siha ( figure 4b ) cells . when cells were treated with tgf-1 , the total number of apoptotic cells ( early apoptotic+apoptotic ) was significantly decreased compared to untreated cells in both cell lines . in cells treated with 50 nm rapamycin , the apoptosis rate significantly increased .
furthermore , the addition of rapamycin significantly abolished the tgf-1-induced anti - apoptosis effects in both cell lines .
to determine whether the emt condition induces an increase in pkm2 , pkm2 expression was detected to compare levels pre- and post - emt state .
cervical cancer cells were stimulated with 10 ng / ml tgf-1 . as showed in figure 1 , siha ( figure 1b ) and hela ( figure 1c ) cells changed morphology from epithelial to fibroblastic - like and spindle - shaped and lost cell - cell contacts , which are characteristics of a mesenchymal - like morphology .
we found that the level of e - cadherin expression was suppressed compared with the pre - emt state .
vimentin and snail family zinc finger 2 ( snai2 ) expressions were increased in the post - emt state ( figure 5 ) .
pkm2 expression was stimulated to increase in the post - emt condition ( figure 5 ) .
the data show that the induction of emt resulted in a decreased level of e - cadherin , and increased vim and pkm2 expression in hela ( figure 5a ) and siha cells ( figure 5b ) .
moreover , atsushiet et al . reported that they were able to knock down pkm2 under emt conditions , but pkm2 knockdown failed to induce spindle - shaped morphological changes .
meanwhile , pkm2 knockdown hindered e - cadherin loss and vim gain compared with the control .
we evaluated the effects of rapamycin ( an mtor inhibitor ) on pkm2 ( a critical glycolytic enzyme ) , and p70s6k ( s6k1 , a downstream effector of mtor ) . to investigate the inhibitory effect on mtor ,
we used 0 , 12.5 , 25 , 50 , and 100 nm rapamycin to treat hela ( figure 6a ) and siha ( figure 6b ) cells for 24 h. ribosomal p70 s6 kinase ( s6k1 ) is a main downstream mtor effector . as shown in figure 6 , rapamycin , a specific mtor inhibitor , inhibited the phosphorylation of p70s6k in a dose - dependent manner .
moreover , we investigated whether mtor / p70s6k signaling decreased pkm2 expression , which is a main downstream p70s6k effector .
we found that rapamycin treatment significantly decreased pkm2 in hela ( figure 6a ) and siha ( figure 6b ) cells .
based on these facts , inhibition of mtor appears to affect cervical cancer through the mtor / p70s6k / pkm2 pathway in cervical cancer cells .
to determine whether rapamycin is involved in regulating emt in cervical cancer , we examined the expression of emt - related markers using western blot analysis .
tgf-1 significantly decreased the expression of e - cadherin and increased the expressions of vimentin , stat3 and snail2 in hela ( figure 7a ) and siha cells ( figure 8a ) .
in addition , at concentration of 50 nm rapamycin reversed tgf-1-induced emt - markers expression by repressing vimentin , stat3 and snail2 expressions and restoring e - cadherin expression in in hela ( figure 7a ) and siha cells ( figure 8a ) .
next , we explored the possible that signaling pathways may be involved . as shown in figures 7 and 8 , we found that tgf-1 significantly increased the phosphorylation of p70s6k , which is the main downstream signaling intermediate of mtor signaling .
simultaneously , tgf-1 significantly increased the expression of pkm2 in hela ( figure 7a ) and siha cells ( figure 8a ) , while rapamycin significantly decreased the expressions of p - p70s6k and pkm2 .
these results suggest that rapamycin reverses tgf-1-induced emt via the mtor / p70s6k / pkm2 pathway .
further , we examined the effect of rapamycin with or without tgf-1 on the morphology of hela and siha cell lines .
after stimulation with 10 ng / ml tgf-1 for 48 h , both hela ( figure 7b ) and siha cells ( figure 8b ) cells became scattered , acquired a spindle - shaped morphology , and lost cell - cell contact , which are characteristics of a mesenchymal - like morphology .
treatment with 50 nm rapamycin for 48 h abolished the tgf-1-induced morphological changes in siha and hela cell lines .
we investigated whether the mechanisms of the mtor pathway affect tumorigenesis in tgf-1-induced emt in cervical cancer cells .
rapamycin , an mtor inhibitor , can inhibit cell proliferation and migration , as well as promoting apoptosis and reversing tgf-1-induced emt via the mtor / p70s6k / pkm2 signaling pathway .
moreover , e - cadherin plays a critical role in maintaining the cell surface and mediating normal epithelial tissue functions .
overexpression of vimentin in cancer correlates well with accelerated tumor growth and invasion . during emt ,
cells start to exhibit a mesenchymal phenotype and show increased vimentin expression , with high motility rates . as shown in figure 1 , we established an emt model in cervical cancer cells .
consistent with these observations , our data indicated that tgf-1-induced cells acquired a striking morphological change and promoted e - cadherin loss and vim gain , as well as promoting proliferation and migration , and inhibiting apoptosis caused by emt .
the mtor signaling pathway plays an important role in integrating intracellular and extracellular signals , and is a central regulator of cell metabolism , growth , proliferation , and survival .
recent studies reported that using rapamycin derivatives in therapy against liver cancer achieves positive outcomes , leading to the implementation of large clinical trials .
pedro et al . showed that rapamycin ( an mtor inhibitor ) inhibits tumor growth in pten - negative ishikawa tumor cells .
our data also showed that inhibition of mtor significantly decreased cell proliferation , apoptosis , and migration , and reverses emt in cervical carcinoma cells .
when cancer cell stayed in the emt state , pkm2 knockdown failed to induce spindle - shaped morphological changes , and hindered e - cadherin reduction and vim increase compared with the control group .
our data show that the induction of emt resulted in a decrease level of e - cadherin , and increased vim and pkm2 expression in cervical cancer cells ( figure 5 ) , suggesting that pkm2 is involved in the emt state .
rps6kb1(p70s6k ) is a key downstream target of mtor , and an essential target for regulating translation rates and additional metabolic processes for cell growth .
pkm2 is a crucial glycolytic enzyme in the oncogenic mtor - induced warburg effect , in which hypoxia - inducible factor-1 ( hif-1 ) and c - myc - hnrnp cascades are the transducers of mtor regulation of pkm2 .
sun et al . reported that mtor upregulation of pyruvate kinase isoenzyme type m2 is critical for aerobic glycolysis and tumor growth .
pkm2 level was augmented in mouse kidney tumors and consequent mtor activation , and was reduced by mtor suppression .
based on these studies on the interaction of mtor , p70s6k , and pkm2 , we investigated whether mtor / p70s6k signaling regulates pkm2 in cells in cervical cancer .
the results showed that the phosphorylation level of p70s6k was inhibited and decreased pkm2 expression , which suggests that the mtor / p70s6k / pkm2 pathway is involved in regulation of cervical cancer ( figure 6 ) .
our data show that tgf-1 significantly increases the expression of p70s6k and pkm2 ( figures 7 , 8) , suggesting the mtor pathway was activated in the process of tgf-1-induced emt , and pkm2 is involved in the process of tgf-1-induced emt .
further , we propose that rapamycin exerts its antitumorigenic effects and abolishes tgf-1-induced emt through mtor / p70s6k / pkm2 signaling in cervical carcinoma cells ( figure 9 ) .
rapamycin , a specific mtor inhibitor , was added to cell cultures to determine whether mtor regulates tgf-1-induced emt .
our data showed that rapamycin treatment significantly decreased phosphorylation of p70s6k and pkm2 in siha and hela cells .
more importantly , we demonstrated that inhibition of mtor / p70s6k / pkm2 signaling is the therapeutic target of cervical cancer .
inhibition of mtor reversed tgf-1-induced emt via the mtor / p70s6k / pkm2 signaling pathway .
we believe this is the first study showing that inhibition of mtor reverses tgf-1-induced emt in tumor cells through mtor / p70s6k / pkm2 pathways .
our data showed that tgf-1 induced proliferation and emt , and rapamycin inhibits cell proliferation and reverses emt .
the mechanism involves suppression of p70s6k activation and reduced pkm2 expression , mediated by inhibiting mtor / p70s6k signaling , which provides novel mechanistic insights into the anti - tumor effects of mtor .
 backgroundepithelial - mesenchymal transition ( emt ) plays an important role in cancer tumorigenesis . transforming growth factor 1 ( tgf-1 ) 
 can induced emt , which could increase tumor migration and invasion . 
 moreover , recent studies have been proven that mammalian target of rapamycin ( mtor ) is a critical regulator of emt . 
 we investigated the mechanisms of mtor in transforming growth factor 1 ( tgf-1)-induced emt in cervical cancer cells.material/methodshela and siha cells were treated with 10 ng / ml tgf-1 to induce emt . 
 then , they were treated with or without rapamycin . 
 cck8 assay was performed to determine cell proliferation . 
 cell migration was detected by wound - healing assay ; apoptosis was analyzed by flow cytometry ; mtor inhibitors inhibited mtor pathway to assess the expression of e - cadherin , vimentin stat3 , snail2 , p - p70s6k , and pkm2 expression.resultstgf-1 promoted proliferation and migration , and attenuated apoptosis in cervical carcinoma cells . 
 rapamycin abolished tgf-1-induced emt cell proliferation and migration and reversed tgf-1-induced emt . 
 e - cadherin were suppressed , whereas vimentin and pkm2 were increased in hela and siha cells after stimulation with tgf-1 . 
 moreover , mtor was activated in the process of tgf-1-induced emt . 
 rapamycin inhibited the phosphorylation of p70s6k . 
 furthermore , inhibition of the mtor pathway decreased pkm2 expression.conclusionsinhibition of the mtor pathway abolished tgf-1-induced emt and reduced mtor / p70s6k signaling , which downregulated pkm2 expression . 
 our results provide novel mechanistic insight into the anti - tumor effects of inhibition of mtor . 
Background
Material and Methods
Cell culture
Cell viability assay
Annexin V-FITC apoptosis assay
Migration assay
Western blot analysis
Statistics
Result
TGF-
Rapamycin inhibits TGF-
The EMT condition induced an increase in PKM2
mTOR is involved in regulation of PKM2 via stimulating mTOR/p70s6k/signaling
Rapamycin reverses EMT in cervical carcinoma cells via regulation of mTOR/p70s6k/PKM2 signaling pathways
Discussion
Conclusions