AIM To clarify the underlying mechanism of formin-like 3(FMNL3)in the promotion of colorectal carcinoma(CRC)cell invasion.METHODS The in vitro biological function analyses of FMNL3 were performed by gain-and loss-of f...AIM To clarify the underlying mechanism of formin-like 3(FMNL3)in the promotion of colorectal carcinoma(CRC)cell invasion.METHODS The in vitro biological function analyses of FMNL3 were performed by gain-and loss-of function approaches.Changes in the F-actin cytoskeleton were detected by the technologies of phalloidin-TRITC labeling and confocal microscopy.The signaling pathway mediated by FMNL3 was explored by western blot,gelatin zymograph assay,co-immunoprecipitation(co-IP),immunofluorescence colocalization,and glutathione S-transferase(GST)pulldown assay.RESULTS The in vitro experimental results showed that FMNL3 significantly promoted the proliferation,invasion,and migration of CRC cells(P<0.05 and P<0.01).Moreover,FMNL3regulated the remodeling of actin-based protrusions such as filopodia and lamellipodia in a RhoC-dependent manner.The western blot and gelatin zymograph assay results indicated that FMNL3 was involved in the RhoC/focal adhesion kinase(FAK)pathway and acted as an effector of RhoC to activate the downstream signaling of p-FAK as well as p-MAPK and p-AKT.This resulted in the increased expression of matrix metalloproteinase 2(MMP2),matrix metalloproteinase 9(MMP9)and vascular endothelial growth factor(VEGF),and the subsequent promotion of CRC cell invasion.The results of TAE226,U0126 or Ly294002 treatment confirmed an essential role of FMNL3 in activation of the RhoC/FAK pathway and the subsequent promotion of CRC invasion.Co-IP,colocalization and GST pull-down assays showed the direct interaction of FMNL3 with RhoC in vivo and in vitro.CONCLUSION FMNL3 regulates the RhoC/FAK signaling pathway and RhoC-dependent remodeling of actin-based protrusions to promote CRC invasion.展开更多
Objective: To study the correlation of FMNL2 and HERC4 expression with the infiltrative growth of cancer cells in cervical cancer tissue. Methods: Patients with cervical cancer who underwent surgical resection in our ...Objective: To study the correlation of FMNL2 and HERC4 expression with the infiltrative growth of cancer cells in cervical cancer tissue. Methods: Patients with cervical cancer who underwent surgical resection in our hospital between July 2014 and September 2016 were selected as the research subjects, the cervical cancer tissue and adjacent tissue were collected from the enrolled patients after operation to extract RNA, and PCR reaction kit was used to determine the mRNA expression of FMNL2, HERC4, epithelial mesenchymal transition genes and proliferation genes. Results: FMNL2, HERC4, Exosomes, Cripto-1, ZEB1, Snail, Vimentin, Piwil2, STOML-2, BCL-2, Smo and XIAP mRNA expression in cervical cancer tissue were significantly higher than those in adjacent tissue;Exosomes, Cripto-1, ZEB1, Snail and Vimentin mRNA expression in cervical cancer tissue with high FMNL2 expression were significantly higher than those in cervical cancer tissue with low FMNL2 expression;Piwil2, STOML-2, BCL-2, Smo and XIAP mRNA expression in cervical cancer tissue with high HERC4 expression were significantly higher than those in cervical cancer tissue with low HERC4 expression. Conclusion: The high expression of FMNL2 and HERC4 in cervical cancer tissue can promote the epithelial mesenchymal transition and proliferation of cancer cells respectively so as to participate in the infiltrative growth of cancer cells.展开更多
[目的]探究食管癌中同源形成素样蛋白2(Formin-like 2,FMNL2)的表达及其与患者预后的关系,分析其在细胞中的生物学功能。[方法]使用免疫组化分析食管癌患者中FMNL2的表达;Kaplan-Meier分析FMNL2的表达与临床预后的关系,Cox回归模型分析F...[目的]探究食管癌中同源形成素样蛋白2(Formin-like 2,FMNL2)的表达及其与患者预后的关系,分析其在细胞中的生物学功能。[方法]使用免疫组化分析食管癌患者中FMNL2的表达;Kaplan-Meier分析FMNL2的表达与临床预后的关系,Cox回归模型分析FMNL2在预后中的价值。使用qRT-PCR和Western blot实验检测细胞中FMNL2的表达;使用CCK8和Transwell小室实验分析FMNL2对细胞增殖、侵袭与迁移的影响。[结果]FMNL2在食管癌组织中表达高于癌旁组织(124.395±18.422 vs 37.876±18.266,P<0.05);FMNL2表达与TNM分期有关(χ^2=6.411,P=0.011);Kaplan-Meier分析显示,与FMNL2低表达组相比,高表达组总生存时间(OS)、无病生存时间(DFS)均明显缩短(47.615 vs 30.125,P=0.01;50.923 vs 41.292,P=0.001);单因素和多因素分析发现FMNL2表达是患者OS的危险因素(HR=3.671,95%CI:1.272~10.591,P=0.016;HR=3.660,95%CI:1.211~11.065,P=0.021);FMNL2在食管癌细胞株中表达高于正常食管上皮细胞;沉默FMNL2能够显著抑制食管癌细胞的增殖、侵袭与迁移能力(P<0.05)。[结论]FMNL2在食管癌发生发展过程中具有重要作用,有望成为食管癌的临床治疗新靶标。展开更多
基金Supported by the National Natural Science Foundation of China,No.81201972the China Postdoctoral Science Foundation,No.2013M531555the Postdoctoral Science Foundation of Jiangxi province,No.2013KY44
文摘AIM To clarify the underlying mechanism of formin-like 3(FMNL3)in the promotion of colorectal carcinoma(CRC)cell invasion.METHODS The in vitro biological function analyses of FMNL3 were performed by gain-and loss-of function approaches.Changes in the F-actin cytoskeleton were detected by the technologies of phalloidin-TRITC labeling and confocal microscopy.The signaling pathway mediated by FMNL3 was explored by western blot,gelatin zymograph assay,co-immunoprecipitation(co-IP),immunofluorescence colocalization,and glutathione S-transferase(GST)pulldown assay.RESULTS The in vitro experimental results showed that FMNL3 significantly promoted the proliferation,invasion,and migration of CRC cells(P<0.05 and P<0.01).Moreover,FMNL3regulated the remodeling of actin-based protrusions such as filopodia and lamellipodia in a RhoC-dependent manner.The western blot and gelatin zymograph assay results indicated that FMNL3 was involved in the RhoC/focal adhesion kinase(FAK)pathway and acted as an effector of RhoC to activate the downstream signaling of p-FAK as well as p-MAPK and p-AKT.This resulted in the increased expression of matrix metalloproteinase 2(MMP2),matrix metalloproteinase 9(MMP9)and vascular endothelial growth factor(VEGF),and the subsequent promotion of CRC cell invasion.The results of TAE226,U0126 or Ly294002 treatment confirmed an essential role of FMNL3 in activation of the RhoC/FAK pathway and the subsequent promotion of CRC invasion.Co-IP,colocalization and GST pull-down assays showed the direct interaction of FMNL3 with RhoC in vivo and in vitro.CONCLUSION FMNL3 regulates the RhoC/FAK signaling pathway and RhoC-dependent remodeling of actin-based protrusions to promote CRC invasion.
文摘Objective: To study the correlation of FMNL2 and HERC4 expression with the infiltrative growth of cancer cells in cervical cancer tissue. Methods: Patients with cervical cancer who underwent surgical resection in our hospital between July 2014 and September 2016 were selected as the research subjects, the cervical cancer tissue and adjacent tissue were collected from the enrolled patients after operation to extract RNA, and PCR reaction kit was used to determine the mRNA expression of FMNL2, HERC4, epithelial mesenchymal transition genes and proliferation genes. Results: FMNL2, HERC4, Exosomes, Cripto-1, ZEB1, Snail, Vimentin, Piwil2, STOML-2, BCL-2, Smo and XIAP mRNA expression in cervical cancer tissue were significantly higher than those in adjacent tissue;Exosomes, Cripto-1, ZEB1, Snail and Vimentin mRNA expression in cervical cancer tissue with high FMNL2 expression were significantly higher than those in cervical cancer tissue with low FMNL2 expression;Piwil2, STOML-2, BCL-2, Smo and XIAP mRNA expression in cervical cancer tissue with high HERC4 expression were significantly higher than those in cervical cancer tissue with low HERC4 expression. Conclusion: The high expression of FMNL2 and HERC4 in cervical cancer tissue can promote the epithelial mesenchymal transition and proliferation of cancer cells respectively so as to participate in the infiltrative growth of cancer cells.
文摘[目的]探究食管癌中同源形成素样蛋白2(Formin-like 2,FMNL2)的表达及其与患者预后的关系,分析其在细胞中的生物学功能。[方法]使用免疫组化分析食管癌患者中FMNL2的表达;Kaplan-Meier分析FMNL2的表达与临床预后的关系,Cox回归模型分析FMNL2在预后中的价值。使用qRT-PCR和Western blot实验检测细胞中FMNL2的表达;使用CCK8和Transwell小室实验分析FMNL2对细胞增殖、侵袭与迁移的影响。[结果]FMNL2在食管癌组织中表达高于癌旁组织(124.395±18.422 vs 37.876±18.266,P<0.05);FMNL2表达与TNM分期有关(χ^2=6.411,P=0.011);Kaplan-Meier分析显示,与FMNL2低表达组相比,高表达组总生存时间(OS)、无病生存时间(DFS)均明显缩短(47.615 vs 30.125,P=0.01;50.923 vs 41.292,P=0.001);单因素和多因素分析发现FMNL2表达是患者OS的危险因素(HR=3.671,95%CI:1.272~10.591,P=0.016;HR=3.660,95%CI:1.211~11.065,P=0.021);FMNL2在食管癌细胞株中表达高于正常食管上皮细胞;沉默FMNL2能够显著抑制食管癌细胞的增殖、侵袭与迁移能力(P<0.05)。[结论]FMNL2在食管癌发生发展过程中具有重要作用,有望成为食管癌的临床治疗新靶标。
