Monitoring the dynamics of cellular pseudopodia at nanoscale has become essential for understanding their diverse and complex functions in living cells.This is made possible by combining single-molecule localization m...Monitoring the dynamics of cellular pseudopodia at nanoscale has become essential for understanding their diverse and complex functions in living cells.This is made possible by combining single-molecule localization microscopy(SMLM)with self-blinking dyes.However,existing self-blinking dyes often face limitations,such as nonspecific blinking and low photostability,which can bring background noise and yield erroneous localization signals,hindering their effectiveness for nanoscale visualization.Here,we present a method for long-term SMLM imaging of cellular pseudopodia dynamics using a blinkogenic probe that exhibits self-blinking activation upon molecular recognition.This approach enabled the precise tracking of various pseudopodia structures,including filopodia,lamellipodia,and(tunneling nanotubes)-nanoscale(TNTs),in living cells.We monitored the growth and fusion of filopodia,as well as the extension and shrinkage of lamellipodia,in real-time.Additionally,we identified two distinct fusion modes between filopodia and lamellipodia and captured the formation of TNTs and their interactions with filopodia,demonstrating the probe's utility in visualizing real-time pseudopodia dynamics at nanoscale.展开更多
Cell adhesion plays an important role in cell physiology. A better understanding of this process could facilitate many clinical therapies. In this study, Rat bone marrow-derived mesenchymal stem cells （rBMSCs） were ...Cell adhesion plays an important role in cell physiology. A better understanding of this process could facilitate many clinical therapies. In this study, Rat bone marrow-derived mesenchymal stem cells （rBMSCs） were cultured on glass substrate, and the morphology and adhesion strength were characterized. The cell morphology was defined as spherical, adhesive, and spreading. The adhesion strengths of the different morphologies exhibited different distribu- tion patterns. The spherical cells exhibited low adhesion strength; the adhesive cells exhibited rapidly increasing adhesion strength while their diameters remained relatively constant. The ad- hesion strength increased with the cell diameter in the spreading cells. These findings suggest that adhesion strength can be quickly assessed by examining the cell morphology.展开更多
目的探讨CDC28蛋白激酶调节亚基2(CKS2)对A2780细胞丝足形成的影响及其可能机制。方法采用慢病毒介导的sh RNA敲除A2780细胞的CKS2基因,显微镜下观察细胞丝足的变化;采用细胞划痕实验检测A2780细胞迁移能力的变化;采用real time PCR检测...目的探讨CDC28蛋白激酶调节亚基2(CKS2)对A2780细胞丝足形成的影响及其可能机制。方法采用慢病毒介导的sh RNA敲除A2780细胞的CKS2基因,显微镜下观察细胞丝足的变化;采用细胞划痕实验检测A2780细胞迁移能力的变化;采用real time PCR检测CKS2敲除对CDC42两种剪接变异体(CDC42-V1和CDC42-V2)表达的影响,以及不同卵巢癌标本中CKS2、CDC42-V1和CDC42-V2剪接变异体的表达情况。结果 CKS2表达抑制后,A2780细胞丝足明显减少,细胞迁移能力明显减弱(P<0.05),CDC42-V1 m RNA表达降低,而CDC42-V2 m RNA表达升高(P<0.05)。Real time PCR结果显示,卵巢癌组织标本中CKS2及CDC42-V1的表达高于对应的正常卵巢组织,而CDC42-V2的表达低于对应的正常卵巢组织(P<0.05)。结论 CKS2通过调控CDC42的选择性剪接而影响细胞丝足的形成,进而影响卵巢癌细胞A2780的迁移能力。展开更多
Intratumoral bacteria,especially Gram-positive bacteria(G^(+)),have a unique bacterial niche in breast cancer that promoted tumor progression.However,the effects of G^(+)have so far been overlooked,serving an“invisib...Intratumoral bacteria,especially Gram-positive bacteria(G^(+)),have a unique bacterial niche in breast cancer that promoted tumor progression.However,the effects of G^(+)have so far been overlooked,serving an“invisible driver”of breast cancer.Moreover,due to the altered biological structure of G^(+)in tumor cells and the penetration barrier of antibiotics,the effect of antibiotic-mediated eradication of G^(+)in tumors is limited.Here,to simultaneously inhibit intratumoral G^(+)and tumor cells via ferroptosis therapy,an amorphous nano-assembly(DFTV)was constructed by assembling doxorubicin(DOX),tannic acid(TA),FeSO_(4),and vancomycin(Van).DFTV treatment effectively targets intratumoral G^(+),thereby inhibiting the growth of the breast tumor and postoperative recurrence by downregulating the expression of inflammatory cytokines,including interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor necrosis factor-alpha(TNF-α).Moreover,inhibiting intracellular G^(+)also restrains the reorganization of F-actin to form pseudopodia,thereby impairing tumor cell motility and blocking metastasis.Collectively,DFTV improves the antitumor efficacy by targeting G^(+)in breast tumors,offering novel insights into overcoming the limitations associated with the lack of intratumoral antibacterial therapy in clinical breast cancer treatment protocols.展开更多
基金supported by the National Natural Science Foundation of China(Nos.22225806,22078314,22278394,22378385)Dalian Institute of Chemical Physics(Nos.DICPI202227,DICPI202436)。
文摘Monitoring the dynamics of cellular pseudopodia at nanoscale has become essential for understanding their diverse and complex functions in living cells.This is made possible by combining single-molecule localization microscopy(SMLM)with self-blinking dyes.However,existing self-blinking dyes often face limitations,such as nonspecific blinking and low photostability,which can bring background noise and yield erroneous localization signals,hindering their effectiveness for nanoscale visualization.Here,we present a method for long-term SMLM imaging of cellular pseudopodia dynamics using a blinkogenic probe that exhibits self-blinking activation upon molecular recognition.This approach enabled the precise tracking of various pseudopodia structures,including filopodia,lamellipodia,and(tunneling nanotubes)-nanoscale(TNTs),in living cells.We monitored the growth and fusion of filopodia,as well as the extension and shrinkage of lamellipodia,in real-time.Additionally,we identified two distinct fusion modes between filopodia and lamellipodia and captured the formation of TNTs and their interactions with filopodia,demonstrating the probe's utility in visualizing real-time pseudopodia dynamics at nanoscale.
基金financial support received from the Sate Key Laboratory of Tribology, Tsinghua University
文摘Cell adhesion plays an important role in cell physiology. A better understanding of this process could facilitate many clinical therapies. In this study, Rat bone marrow-derived mesenchymal stem cells （rBMSCs） were cultured on glass substrate, and the morphology and adhesion strength were characterized. The cell morphology was defined as spherical, adhesive, and spreading. The adhesion strengths of the different morphologies exhibited different distribu- tion patterns. The spherical cells exhibited low adhesion strength; the adhesive cells exhibited rapidly increasing adhesion strength while their diameters remained relatively constant. The ad- hesion strength increased with the cell diameter in the spreading cells. These findings suggest that adhesion strength can be quickly assessed by examining the cell morphology.
文摘目的探讨CDC28蛋白激酶调节亚基2(CKS2)对A2780细胞丝足形成的影响及其可能机制。方法采用慢病毒介导的sh RNA敲除A2780细胞的CKS2基因,显微镜下观察细胞丝足的变化;采用细胞划痕实验检测A2780细胞迁移能力的变化;采用real time PCR检测CKS2敲除对CDC42两种剪接变异体(CDC42-V1和CDC42-V2)表达的影响,以及不同卵巢癌标本中CKS2、CDC42-V1和CDC42-V2剪接变异体的表达情况。结果 CKS2表达抑制后,A2780细胞丝足明显减少,细胞迁移能力明显减弱(P<0.05),CDC42-V1 m RNA表达降低,而CDC42-V2 m RNA表达升高(P<0.05)。Real time PCR结果显示,卵巢癌组织标本中CKS2及CDC42-V1的表达高于对应的正常卵巢组织,而CDC42-V2的表达低于对应的正常卵巢组织(P<0.05)。结论 CKS2通过调控CDC42的选择性剪接而影响细胞丝足的形成,进而影响卵巢癌细胞A2780的迁移能力。
基金supported by the National Natural Science Foundation of China(No.82173754).
文摘Intratumoral bacteria,especially Gram-positive bacteria(G^(+)),have a unique bacterial niche in breast cancer that promoted tumor progression.However,the effects of G^(+)have so far been overlooked,serving an“invisible driver”of breast cancer.Moreover,due to the altered biological structure of G^(+)in tumor cells and the penetration barrier of antibiotics,the effect of antibiotic-mediated eradication of G^(+)in tumors is limited.Here,to simultaneously inhibit intratumoral G^(+)and tumor cells via ferroptosis therapy,an amorphous nano-assembly(DFTV)was constructed by assembling doxorubicin(DOX),tannic acid(TA),FeSO_(4),and vancomycin(Van).DFTV treatment effectively targets intratumoral G^(+),thereby inhibiting the growth of the breast tumor and postoperative recurrence by downregulating the expression of inflammatory cytokines,including interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor necrosis factor-alpha(TNF-α).Moreover,inhibiting intracellular G^(+)also restrains the reorganization of F-actin to form pseudopodia,thereby impairing tumor cell motility and blocking metastasis.Collectively,DFTV improves the antitumor efficacy by targeting G^(+)in breast tumors,offering novel insights into overcoming the limitations associated with the lack of intratumoral antibacterial therapy in clinical breast cancer treatment protocols.
