Background:Bladder cancer,characterized by a high potential of tumor recurrence,has high lifelong monitoring and treatment costs.To date,tumor cells with intrinsic softness have been identified to function as cancer s...Background:Bladder cancer,characterized by a high potential of tumor recurrence,has high lifelong monitoring and treatment costs.To date,tumor cells with intrinsic softness have been identified to function as cancer stem cells in several cancer types.Nonetheless,the existence of soft tumor cells in bladder tumors remains elusive.Thus,our study aimed to develop a microbarrier microfluidic chip to efficiently isolate deformable tumor cells from distinct types of bladder cancer cells.Methods:The stiffness of bladder cancer cells was determined by atomic force microscopy(AFM).The modified microfluidic chip was utilized to separate soft cells,and the 3D Matrigel culture system was to maintain the softness of tumor cells.Expression patterns of integrinβ8(ITGB8),protein kinase B(AKT),and mammalian target of rapamycin(mTOR)were determined by Western blotting.Double immunostaining was conducted to examine the interaction between F-actin and tripartite motif containing 59(TRIM59).The stem-cell-like characteristics of soft cells were explored by colony formation assay and in vivo studies upon xenografted tumor models.Results:Using our newly designed microfluidic approach,we identified a small fraction of soft tumor cells in bladder cancer cells.More importantly,the existence of soft tumor cells was confirmed in clinical human bladder cancer specimens,in which the number of soft tumor cells was associated with tumor relapse.Furthermore,we demonstrated that the biomechanical stimuli arising from 3D Matrigel activated the F-actin/ITGB8/TRIM59/AKT/mTOR/glycolysis pathways to enhance the softness and tumorigenic capacity of tumor cells.Simultaneously,we detected a remarkable up-regulation in ITGB8,TRIM59,and phospho-AKT in clinical bladder recurrent tumors compared with their non-recurrent counterparts.Conclusions:The ITGB8/TRIM59/AKT/mTOR/glycolysis axis plays a crucial role in modulating tumor softness and stemness.Meanwhile,the soft tumor cells become more sensitive to chemotherapy after stiffening,that offers new insights for hampering tumor progression and recurrence.展开更多
Background:Diminished sensitivity towards chemotherapy remains the major impediment to the clinical treatment of bladder cancer.However,the critical elements in control of chemotherapy resistance remain obscure.Method...Background:Diminished sensitivity towards chemotherapy remains the major impediment to the clinical treatment of bladder cancer.However,the critical elements in control of chemotherapy resistance remain obscure.Methods:We adopted improved collagen gels and performed cytotoxicity analysis of doxorubicin(DOX)and mitomycin C(MMC)of bladder cancer cells in a 3D culture system.We then detected the expression of multidrug resistant gene ABCB1,dormancy-associated functional protein chicken ovalbumin upstream-transcription factor 1(COUPTF1),cell proliferation marker Ki-67,and cellular senescence marker senescence-associatedβ-galactosidase(SA-β-Gal)in these cells.We further tested the effects of integrin blockade or protein kinase B(AKT)inhibitor on the senescent state of bladder cancer.Also,we examined the tumor growth and survival time of bladder cancer mouse models given the combination treatment of chemotherapeutic agents and integrinα2β1 ligand peptide TFA(TFA).Results:Collagen gels played a repressive role in bladder cancer cell apoptosis induced by DOX and MMC.In mechanism,collagen activated the integrinβ1/AKT cascade to drive bladder cancer cells into a premature senescence state via the p21/p53 pathway,thus attenuating chemotherapy-induced apoptosis.In addition,TFA had the ability to mediate the switch from senescence to apoptosis of bladder cancer cells in xenograft mice.Meanwhile,TFA combined with chemotherapeutic drugs produced a substantial suppression of tumor growth as well as an extension of survival time in vivo.Conclusions:Based on our finding that integrinβ1/AKT acted primarily to impart premature senescence to bladder cancer cells cultured in collagen gel,we suggest that integrinβ1 might be a feasible target for bladder cancer eradication.展开更多
The extracellular matrix(ECM)serves as signals that regulate specific cell states in tumor tissues.Increasing evidence suggests that extracellular biomechanical force signals are critical in tumor progression.In this ...The extracellular matrix(ECM)serves as signals that regulate specific cell states in tumor tissues.Increasing evidence suggests that extracellular biomechanical force signals are critical in tumor progression.In this study,we aimed to explore the influence of ECM-derived biomechanical force on breast cancer cell status.Experiments were conducted using 3D collagen,fibrinogen.展开更多
To the Editor:Testosterone is a fundamental male sex hormone produced by the testicular Leydig cells.Testosterone levels are affected by age,peaking in the 20s and 30s and gradually declining thereafter.[1]Low testost...To the Editor:Testosterone is a fundamental male sex hormone produced by the testicular Leydig cells.Testosterone levels are affected by age,peaking in the 20s and 30s and gradually declining thereafter.[1]Low testosterone level may predispose men,especially older men,to a poor prognosis or death in coronavirus disease 2019(COVID-19).[2]Therefore,testosterone levels have a significant impact on the health status of older men.The identification of modifiable non-drug factors affecting testosterone levels in older men is important for improving their health.展开更多
基金supported by the National Natural Science Foundation of China(Nos.81902578,81974098,8197032158)China Postdoctoral Science Foundation(No.2020M670057ZX)+3 种基金Programs from Science and Technology Department of Sichuan Province(No.2021YJ0462)Post-doctoral Science Research Foundation of Sichuan University(No.2020SCU12041)Post-Doctor Research Project,West China Hospital,Sichuan University(Nos.2018HXBH084,2019HXBH092)the National key research and development program of China(No.2020YFC2008601)
文摘Background:Bladder cancer,characterized by a high potential of tumor recurrence,has high lifelong monitoring and treatment costs.To date,tumor cells with intrinsic softness have been identified to function as cancer stem cells in several cancer types.Nonetheless,the existence of soft tumor cells in bladder tumors remains elusive.Thus,our study aimed to develop a microbarrier microfluidic chip to efficiently isolate deformable tumor cells from distinct types of bladder cancer cells.Methods:The stiffness of bladder cancer cells was determined by atomic force microscopy(AFM).The modified microfluidic chip was utilized to separate soft cells,and the 3D Matrigel culture system was to maintain the softness of tumor cells.Expression patterns of integrinβ8(ITGB8),protein kinase B(AKT),and mammalian target of rapamycin(mTOR)were determined by Western blotting.Double immunostaining was conducted to examine the interaction between F-actin and tripartite motif containing 59(TRIM59).The stem-cell-like characteristics of soft cells were explored by colony formation assay and in vivo studies upon xenografted tumor models.Results:Using our newly designed microfluidic approach,we identified a small fraction of soft tumor cells in bladder cancer cells.More importantly,the existence of soft tumor cells was confirmed in clinical human bladder cancer specimens,in which the number of soft tumor cells was associated with tumor relapse.Furthermore,we demonstrated that the biomechanical stimuli arising from 3D Matrigel activated the F-actin/ITGB8/TRIM59/AKT/mTOR/glycolysis pathways to enhance the softness and tumorigenic capacity of tumor cells.Simultaneously,we detected a remarkable up-regulation in ITGB8,TRIM59,and phospho-AKT in clinical bladder recurrent tumors compared with their non-recurrent counterparts.Conclusions:The ITGB8/TRIM59/AKT/mTOR/glycolysis axis plays a crucial role in modulating tumor softness and stemness.Meanwhile,the soft tumor cells become more sensitive to chemotherapy after stiffening,that offers new insights for hampering tumor progression and recurrence.
基金supported by the National Natural Science Foundation of China(Grants No.81902578,81974098,and 8197032158)the National Key Research and Development Program of China(Grants No.2017YFC0908003 and 2017YFC0908004)+2 种基金the Project of Health Commission of Sichuan Province(Grant No.20PJ062)Post-doctoral Science Research Foundation of Sichuan University(Grant No.2020SCU12041)Post-doctor Research Project,West China Hospital,Sichuan University(Grant No.2018HXBH084).
文摘Background:Diminished sensitivity towards chemotherapy remains the major impediment to the clinical treatment of bladder cancer.However,the critical elements in control of chemotherapy resistance remain obscure.Methods:We adopted improved collagen gels and performed cytotoxicity analysis of doxorubicin(DOX)and mitomycin C(MMC)of bladder cancer cells in a 3D culture system.We then detected the expression of multidrug resistant gene ABCB1,dormancy-associated functional protein chicken ovalbumin upstream-transcription factor 1(COUPTF1),cell proliferation marker Ki-67,and cellular senescence marker senescence-associatedβ-galactosidase(SA-β-Gal)in these cells.We further tested the effects of integrin blockade or protein kinase B(AKT)inhibitor on the senescent state of bladder cancer.Also,we examined the tumor growth and survival time of bladder cancer mouse models given the combination treatment of chemotherapeutic agents and integrinα2β1 ligand peptide TFA(TFA).Results:Collagen gels played a repressive role in bladder cancer cell apoptosis induced by DOX and MMC.In mechanism,collagen activated the integrinβ1/AKT cascade to drive bladder cancer cells into a premature senescence state via the p21/p53 pathway,thus attenuating chemotherapy-induced apoptosis.In addition,TFA had the ability to mediate the switch from senescence to apoptosis of bladder cancer cells in xenograft mice.Meanwhile,TFA combined with chemotherapeutic drugs produced a substantial suppression of tumor growth as well as an extension of survival time in vivo.Conclusions:Based on our finding that integrinβ1/AKT acted primarily to impart premature senescence to bladder cancer cells cultured in collagen gel,we suggest that integrinβ1 might be a feasible target for bladder cancer eradication.
基金This work was supported by following fundings:CAMS Innovation Fund for Medical Sciences(CIFMS)2021-I2M-1-014the National Natural Science Foundation of China(Grant No.81902578,81974098,8197032158)+3 种基金the National key research and development program of China(Grant No.2017YFC0908003,2017YFC0908004)the Project of Health Commission of Sichuan Province(20PJ062)Post-doctoral Science Research Foundation of Sichuan University(2020SCU12041)Post-Doctor Research Project,West China Hospital,Sichuan University(2018HXBH084).
文摘The extracellular matrix(ECM)serves as signals that regulate specific cell states in tumor tissues.Increasing evidence suggests that extracellular biomechanical force signals are critical in tumor progression.In this study,we aimed to explore the influence of ECM-derived biomechanical force on breast cancer cell status.Experiments were conducted using 3D collagen,fibrinogen.
基金National Key Research and Development Program of China(No.2017YFC0908003)National Natural Science Foundation of China(Nos.81902578 and 81974098)+3 种基金China Post-doctoral Science Foundation(No.2017M612971)Post-doctoral Science Research Foundation of Sichuan University(No.2020SCU12041)Post-doctoral Research Project,West China Hospital,Sichuan University(No.2018HXBH085)National Clinical Research Center for Geriatrics,West China Hospital,Sichuan University(No.Z2018C01)
文摘To the Editor:Testosterone is a fundamental male sex hormone produced by the testicular Leydig cells.Testosterone levels are affected by age,peaking in the 20s and 30s and gradually declining thereafter.[1]Low testosterone level may predispose men,especially older men,to a poor prognosis or death in coronavirus disease 2019(COVID-19).[2]Therefore,testosterone levels have a significant impact on the health status of older men.The identification of modifiable non-drug factors affecting testosterone levels in older men is important for improving their health.