[目的]探索CLIC4/ARF6信号通路对SiHa细胞增殖、侵袭能力的影响。[方法]用蛋白免疫印迹实验检测HUCEC细胞、SiHa细胞、C33A细胞以及HeLa细胞中CLIC4与ARF6的蛋白表达水平;将人宫颈癌SiHa细胞分为3组:si NC组、si CLIC4组与si ARF6组。通...[目的]探索CLIC4/ARF6信号通路对SiHa细胞增殖、侵袭能力的影响。[方法]用蛋白免疫印迹实验检测HUCEC细胞、SiHa细胞、C33A细胞以及HeLa细胞中CLIC4与ARF6的蛋白表达水平;将人宫颈癌SiHa细胞分为3组:si NC组、si CLIC4组与si ARF6组。通过CCK-8实验检测SiHa细胞的增殖活性;Transwell实验检测SiHa细胞的侵袭数量;通过TUNEL实验检测SiHa细胞的凋亡率。[结果]与HUCEC细胞相比,SiHa细胞、C33A细胞及HeLa细胞的CLIC4与ARF6的蛋白表达增加(0.13±0.02 vs 0.92±0.11 vs 0.89±0.08 vs 0.87±0.05;0.21±0.05 vs 0.72±0.06 vs 0.81±0.03 vs 0.77±0.03)。与si NC组比较,si CLIC4组及si ARF6组的SiHa细胞的增殖活性降低(P<0.05);si CLIC4组、si ARF6组的SiHa细胞侵袭数量减少(85.23±9.22 vs 36.62±7.75 vs 39.83±11.58);si CLIC4组、si ARF6组的SiHa细胞的凋亡率增加(5.73%±0.76%vs 26.58%±0.29%vs 29.03%±0.58%)。[结论]CLIC4与ARF6在宫颈癌细胞中表达上调,下调CLIC4或ARF6的表达后,宫颈癌SiHa细胞的增殖活性与侵袭数量降低。此外,下调CLIC4或ARF6的表达后,宫颈癌SiHa细胞凋亡率增加。展开更多
ADP-ribosylation factor 6(Arf6),a small G-protein of the Ras superfamily,plays pivotal roles in multiple cellular events,including exocytosis,endocytosis,actin remodeling,plasma membrane reorganization and vesicular t...ADP-ribosylation factor 6(Arf6),a small G-protein of the Ras superfamily,plays pivotal roles in multiple cellular events,including exocytosis,endocytosis,actin remodeling,plasma membrane reorganization and vesicular transport.Arf6 regulates the progression of cancer through the activation of cell motility and invasion.Aberrant Arf6 activation is a potential therapeutic target.This review aims to understand the comprehensive function of Arf6 for future cancer therapy.The Arf6 GEFs,protein structure,and roles in cancer have been summarized.Comprehending the mechanism underlying Arf6-mediated cancer cell growth and survival is essential.The structural features of Arf6 and its efforts are discussed and may be contributed to the discovery of future novel protein-protein interaction inhibitors.In addition,Arf6 inhibitors and mechanism of action are listed in the table.This review further emphasizes the crucial roles in drug resistance and attempts to offer an outlook of Arf6 in cancer therapy.展开更多
Cell migration requires the generation of branched actin networks and recruitment of vesicular membrane that power the protrusion of the plasma membrane in lamellipodia.However,the molecular mechanisms underlying dyna...Cell migration requires the generation of branched actin networks and recruitment of vesicular membrane that power the protrusion of the plasma membrane in lamellipodia.However,the molecular mechanisms underlying dynamic recruitment of vesicular membrane during cell migration remain elusive.Here,we report a critical mechanism underlying epidermal growth factor(EGF)-elicited Akt signaling-steered cell migration.Using functional proteomics screen,we identified a novel ADP-ribosylation factor 6(ARF6)–ACAP4 signaling regulator,Acapin,which inhibits the GTPase-activating protein(GAP)activity of ACAP4 to activate ARF6 GTPase in vitro.In cells,EGF stimulation elicits Akt signaling,which recruits Acapin to the lamellipodium membrane via phosphorylation of Acapin at its Ser247 residue and enhances the binding of Acapin to ACAP4 to elevate the ARF6-GTP level.Therefore,Acapin is required for efficiently stimulating cell migration by EGF–Akt signaling.Together,our results demonstrate the role of Acapin in relaying the Akt signaling cascade during cell migration processes.展开更多
文摘[目的]探索CLIC4/ARF6信号通路对SiHa细胞增殖、侵袭能力的影响。[方法]用蛋白免疫印迹实验检测HUCEC细胞、SiHa细胞、C33A细胞以及HeLa细胞中CLIC4与ARF6的蛋白表达水平;将人宫颈癌SiHa细胞分为3组:si NC组、si CLIC4组与si ARF6组。通过CCK-8实验检测SiHa细胞的增殖活性;Transwell实验检测SiHa细胞的侵袭数量;通过TUNEL实验检测SiHa细胞的凋亡率。[结果]与HUCEC细胞相比,SiHa细胞、C33A细胞及HeLa细胞的CLIC4与ARF6的蛋白表达增加(0.13±0.02 vs 0.92±0.11 vs 0.89±0.08 vs 0.87±0.05;0.21±0.05 vs 0.72±0.06 vs 0.81±0.03 vs 0.77±0.03)。与si NC组比较,si CLIC4组及si ARF6组的SiHa细胞的增殖活性降低(P<0.05);si CLIC4组、si ARF6组的SiHa细胞侵袭数量减少(85.23±9.22 vs 36.62±7.75 vs 39.83±11.58);si CLIC4组、si ARF6组的SiHa细胞的凋亡率增加(5.73%±0.76%vs 26.58%±0.29%vs 29.03%±0.58%)。[结论]CLIC4与ARF6在宫颈癌细胞中表达上调,下调CLIC4或ARF6的表达后,宫颈癌SiHa细胞的增殖活性与侵袭数量降低。此外,下调CLIC4或ARF6的表达后,宫颈癌SiHa细胞凋亡率增加。
基金the National Natural Science Foundation of China(NSFC)(No.81773594,82204224)Chunhui Program-Cooperative Research Project of the Ministry of Education+3 种基金Liaoning Province Natural Science Foundation(No.2022-MS241,China)China Postdoctoral Science Foundation(No.2021M693957,China)Shenyang Young and Middle-aged Innovative Talents Support Program(RC210446,China)Project of the Educational Department of Liaoning Province(No.LJKZ0919,China),for financial supports。
文摘ADP-ribosylation factor 6(Arf6),a small G-protein of the Ras superfamily,plays pivotal roles in multiple cellular events,including exocytosis,endocytosis,actin remodeling,plasma membrane reorganization and vesicular transport.Arf6 regulates the progression of cancer through the activation of cell motility and invasion.Aberrant Arf6 activation is a potential therapeutic target.This review aims to understand the comprehensive function of Arf6 for future cancer therapy.The Arf6 GEFs,protein structure,and roles in cancer have been summarized.Comprehending the mechanism underlying Arf6-mediated cancer cell growth and survival is essential.The structural features of Arf6 and its efforts are discussed and may be contributed to the discovery of future novel protein-protein interaction inhibitors.In addition,Arf6 inhibitors and mechanism of action are listed in the table.This review further emphasizes the crucial roles in drug resistance and attempts to offer an outlook of Arf6 in cancer therapy.
基金supported by grants from the Ministry of Science and Technology of China and the National Natural Science Foundationof China(2022YFA1303100,32090040,92254302,91953000,32300649,32400593,32170737,31501130,91854203,31621002,and 2017YFA0503600)the Ministry of Education(IRT_17R102)+1 种基金the Fundamental Research Funds for the Central Universities(KB910000oo07,KB9100000006,and KB9100000013)Plans for Major Provincial Science&Technology Projects of Anhui Province(202303a0702003).
文摘Cell migration requires the generation of branched actin networks and recruitment of vesicular membrane that power the protrusion of the plasma membrane in lamellipodia.However,the molecular mechanisms underlying dynamic recruitment of vesicular membrane during cell migration remain elusive.Here,we report a critical mechanism underlying epidermal growth factor(EGF)-elicited Akt signaling-steered cell migration.Using functional proteomics screen,we identified a novel ADP-ribosylation factor 6(ARF6)–ACAP4 signaling regulator,Acapin,which inhibits the GTPase-activating protein(GAP)activity of ACAP4 to activate ARF6 GTPase in vitro.In cells,EGF stimulation elicits Akt signaling,which recruits Acapin to the lamellipodium membrane via phosphorylation of Acapin at its Ser247 residue and enhances the binding of Acapin to ACAP4 to elevate the ARF6-GTP level.Therefore,Acapin is required for efficiently stimulating cell migration by EGF–Akt signaling.Together,our results demonstrate the role of Acapin in relaying the Akt signaling cascade during cell migration processes.
