BACKGROUND Hepatocellular carcinoma(HCC)is a major health challenge with high incidence and poor survival rates in China.Systemic therapies,particularly tyrosine kinase inhibitors(TKIs),are the first-line treatment fo...BACKGROUND Hepatocellular carcinoma(HCC)is a major health challenge with high incidence and poor survival rates in China.Systemic therapies,particularly tyrosine kinase inhibitors(TKIs),are the first-line treatment for advanced HCC,but resistance is common.The Rho GTPase family member Rho GTPase activating protein 12(ARHGAP12),which regulates cell adhesion and invasion,is a potential therapeutic target for overcoming TKI resistance in HCC.However,no studies on the expression of ARHGAP12 in HCC and its role in resistance to TKIs have been reported.AIM To unveil the expression of ARHGAP12 in HCC,its role in TKI resistance and its potential associated pathways.METHODS This study used single-cell RNA sequencing(scRNA-seq)to evaluate ARHGAP12 mRNA levels and explored its mechanisms through enrichment analysis.CellChat was used to investigate focal adhesion(FA)pathway regulation.We integrated bulk RNA data(RNA-seq and microarray),immunohistochemistry and proteomics to analyze ARHGAP12 mRNA and protein levels,correlating with clinical outcomes.We assessed ARHGAP12 expression in TKI-resistant HCC,integrated conventional HCC to explore its mechanism,identified intersecting FA pathway genes with scRNA-seq data and evaluated its response to TKI and immunotherapy.RESULTS ARHGAP12 mRNA was found to be highly expressed in malignant hepatocytes and to regulate FA.In malignant hepatocytes in high-score FA groups,MDK-[integrin alpha 6(ITGA6)+integrinβ-1(ITGB1)]showed specificity in ligand-receptor interactions.ARHGAP12 mRNA and protein were upregulated in bulk RNA,immunohistochemistry and proteomics,and higher expression was associated with a worse prognosis.ARHGAP12 was also found to be a TKI resistance gene that regulated the FA pathway.ITGB1 was identified as a crossover gene in the FA pathway in both scRNA-seq and bulk RNA.High expression of ARHGAP12 was associated with adverse reactions to sorafenib,cabozantinib and regorafenib,but not to immunotherapy.CONCLUSION ARHGAP12 expression is elevated in HCC and TKI-resistant HCC,and its regulatory role in FA may underlie the TKI-resistant phenotype.展开更多
Focal adhesion kinase(FAK)is a 125-kDa non-receptor protein tyrosine.Growth factors or the clustering of integrins facilitate the rapid phosphorylation of FAK at Tyr-397 and this in turn recruits Src-family protein ty...Focal adhesion kinase(FAK)is a 125-kDa non-receptor protein tyrosine.Growth factors or the clustering of integrins facilitate the rapid phosphorylation of FAK at Tyr-397 and this in turn recruits Src-family protein tyrosine kinases,resulting in the phosphorylation of Tyr-576 and Tyr-577 in the FAK activation loop and full catalytic FAK activation.FAK plays a critical role in the biological processes of normal and cancer cells including the gastrointestinal tract.FAK also plays an important role in the restitution,cell survival and apoptosis and carcinogenesis of the gastrointestinal tract.FAK is over-expressed in cancer cells and its over-expression and elevated activities are associated with motility and invasion of cancer cells.FAK has been proposed as a potential target in cancer therapy.Small molecule inhibitors effectively inhibit the kinase activity of FAK and show a potent inhibitory effect for the proliferation and migration of tumor cells,indicating a high potential for application in cancer therapy.展开更多
Focal adhesion kinase(FAK) is a major integrin- dep-endent tyrosine phosphorylated protein, recently, FAK association with colorectal cancer(CRC) has gained at-tention. The various cancer-promoting mechanisms that ass...Focal adhesion kinase(FAK) is a major integrin- dep-endent tyrosine phosphorylated protein, recently, FAK association with colorectal cancer(CRC) has gained at-tention. The various cancer-promoting mechanisms that associated with FAK can be implicated in the progression of CRC. The interactions between structural features of FAK and various kinases could be closely related to growth, survival, and metastasis in CRC cells. These interactions include human epithelial growth factor re-ceptor, c-Met, platelet-derived growth factor receptor, vascular endothelial growth factor receptor, and Src. Such interactions can trigger the survival signaling of CRC cells and are also involved signaling downstream of phosphatidylinositol 3-kinase, AKT, and the extracellular regulated kinase. Based on this scientific background, many pharmaceutical companies are taking efforts to develop FAK inhibitors to treat solid cancer including CRC. Although the anti-cancer efficacies have been noted in many studies, the commercial drugs have not been deve-loped yet. Therefore, the FAK research on CRC is expec-ted to gain momentum and be highly appreciated as a potential field for developing the new drugs. Therefore, the studies on FAK that effect on the progression of human CRC s would be possible to suggest various app-roaches to CRC treatment, and FAK could be a potential target as an anticancer candidate for CRC therapies.展开更多
动脉粥样硬化（AS）是一类主要累及大中动脉血管壁的慢性炎症疾病,主要由血脂紊乱引起,炎症反应参与AS发生、发展的全过程.在AS发生发展过程中,炎症细胞、血管内皮细胞、平滑肌细胞和细胞因子、趋化蛋白、黏附分子等相互作用,相互影响...动脉粥样硬化（AS）是一类主要累及大中动脉血管壁的慢性炎症疾病,主要由血脂紊乱引起,炎症反应参与AS发生、发展的全过程.在AS发生发展过程中,炎症细胞、血管内皮细胞、平滑肌细胞和细胞因子、趋化蛋白、黏附分子等相互作用,相互影响动脉粥样化过程.促炎细胞因子可改变动脉粥样硬化早期血管内皮功能,诱导血管内皮细胞表达趋化因子和黏附分子,促进白细胞、淋巴细胞和单核细胞迁移、募集、黏附到发炎的血管壁中.白细胞在动脉血管内膜中被局部产生的细胞因子永久激活,其可以通过刺激清道夫受体的表达和增强细胞介导的氧化来加速巨噬细胞向泡沫细胞的转化,加剧动脉粥样硬化病变进展.致炎细胞因子发挥生物学功能是通过与细胞膜表面受体相互作用,经过跨膜信号转导活化细胞内的相关信号通路,最终促进靶基因的表达.目前认为,丝裂原活化蛋白激酶（mitogen activated protein kinase,MAPKs）、Janus激酶-信号转导子及转录激活子（Janus kinase sig-nal transduction and activator of transcription,JAK-STAT）和核因子kB（nuclear factor kB,NF-kB）是细胞内3条重要信号通路,在炎症信号转导调控中起重要作用.MAPKs是细胞外刺激信号转导至细胞内及核内并引起生物化学功能如细胞增殖、分化、转化及凋亡等的主要信号转导通路.展开更多
AIM:To investigate the effects and mechanism of disruption of focal adhesion kinase(FAK) expression on collagen metabolism in rat hepatic stellate cells(HSC).METHODS:The plasmids expressing FAK short hairpin RNA(shRNA...AIM:To investigate the effects and mechanism of disruption of focal adhesion kinase(FAK) expression on collagen metabolism in rat hepatic stellate cells(HSC).METHODS:The plasmids expressing FAK short hairpin RNA(shRNA) were transfected into HSC-T6 cells,and the level of FAK expression was determined by both real-time quantitative polymerase chain reaction(QPCR) and Western blotting analysis.The production of type collagen and type collagen in FAK-disrupted cells was analyzed by real-time Q-PCR.The level of collagen metabolism proteins,including matrix metalloproteinases-13(MMP-13) and tissue inhibitors of metalloproteinases-1(TIMP-1) was also determined by both real-time Q-PCR and Western blotting analysis.RESULTS:The transfection of FAK shRNA plasmids into HSC resulted in disrupted FAK expression.Compared with the HK group,the levels of type collagen and type collagen mRNA transcripts in FAK shRNA plas-mid group were signif icantly decreased(0.69 ± 0.03 vs 1.96 ± 0.15,P = 0.000;0.59 ± 0.07 vs 1.62 ± 0.12,P = 0.020).The production of TIMP-1 in this cell type was also signif icantly reduced at both mRNA and protein levels(0.49 ± 0.02 vs 1.72 ± 0.10,P = 0.005;0.76 ± 0.08 vs 2.31 ± 0.24,P = 0.000).However,the expression of MMP-13 mRNA could be significantly up-regulated by the transfection of FAK shRNA plasmids into HSC(1.74 ± 0.20 vs 1.09 ± 0.09,P = 0.000).CONCLUSION:These data support the hypothesis that shRNA-mediated disruption of FAK expression could attenuate extracellular matrix(ECM) synthesis and promote ECM degradation,making FAK a potential target for novel anti-f ibrosis therapies.展开更多
目的:本研究旨在证实朝鲜白头翁、人参和甘草的复合水提物(water extract ofPulsatillakoreana(Yabe ex Nakai)Nakai ex T.Mori.,PanaxginsengC.A.Meyer andGlycyrrhizauralensisFisch,WEPPG)的抗血管生成作用。方法:使用纤维母细胞生...目的:本研究旨在证实朝鲜白头翁、人参和甘草的复合水提物(water extract ofPulsatillakoreana(Yabe ex Nakai)Nakai ex T.Mori.,PanaxginsengC.A.Meyer andGlycyrrhizauralensisFisch,WEPPG)的抗血管生成作用。方法:使用纤维母细胞生长因子致血管生成的人类脐静脉内皮细胞模型衡量细胞的增殖、黏附及迁移,同时进行细管形成实验及纤维母细胞生长因子致鸡胚绒毛尿囊膜血管生成实验检测WEPPG的抗血管生成作用。结果:WEPPG能够显著抑制纤维母细胞生长因子所致血管生成的人类脐静脉内皮细胞的增殖、黏附及迁移。信号蛋白分析显示多种蛋白表达变化,如细胞周期素A、p63、KIP2的上调及nibrin蛋白和黏着斑激酶的下调。与对照组相比,WEPPG显著减少了鸡胚绒毛尿囊膜血管生成。结论:本研究的结果证实了WEPPG的抗血管生成作用,这可能是这种药物具有抗癌功效的原因之一。展开更多
基金Supported by National Natural Science Foundation of China,No.82260581Guangxi Zhuang Autonomous Region Health Committee Scientific Research Project,No.Z20201147+3 种基金Guangxi Medical University Education and Teaching Reform Project,No.2021XJGA02Undergraduate Teaching Reform Project of Guangxi Higher Education,No.2023JGB163Guangxi Medical University Teacher Teaching Ability Development Project,No.2202JFA20China Undergraduate Innovation and Entrepreneurship Training Program,No.S202310598170.
文摘BACKGROUND Hepatocellular carcinoma(HCC)is a major health challenge with high incidence and poor survival rates in China.Systemic therapies,particularly tyrosine kinase inhibitors(TKIs),are the first-line treatment for advanced HCC,but resistance is common.The Rho GTPase family member Rho GTPase activating protein 12(ARHGAP12),which regulates cell adhesion and invasion,is a potential therapeutic target for overcoming TKI resistance in HCC.However,no studies on the expression of ARHGAP12 in HCC and its role in resistance to TKIs have been reported.AIM To unveil the expression of ARHGAP12 in HCC,its role in TKI resistance and its potential associated pathways.METHODS This study used single-cell RNA sequencing(scRNA-seq)to evaluate ARHGAP12 mRNA levels and explored its mechanisms through enrichment analysis.CellChat was used to investigate focal adhesion(FA)pathway regulation.We integrated bulk RNA data(RNA-seq and microarray),immunohistochemistry and proteomics to analyze ARHGAP12 mRNA and protein levels,correlating with clinical outcomes.We assessed ARHGAP12 expression in TKI-resistant HCC,integrated conventional HCC to explore its mechanism,identified intersecting FA pathway genes with scRNA-seq data and evaluated its response to TKI and immunotherapy.RESULTS ARHGAP12 mRNA was found to be highly expressed in malignant hepatocytes and to regulate FA.In malignant hepatocytes in high-score FA groups,MDK-[integrin alpha 6(ITGA6)+integrinβ-1(ITGB1)]showed specificity in ligand-receptor interactions.ARHGAP12 mRNA and protein were upregulated in bulk RNA,immunohistochemistry and proteomics,and higher expression was associated with a worse prognosis.ARHGAP12 was also found to be a TKI resistance gene that regulated the FA pathway.ITGB1 was identified as a crossover gene in the FA pathway in both scRNA-seq and bulk RNA.High expression of ARHGAP12 was associated with adverse reactions to sorafenib,cabozantinib and regorafenib,but not to immunotherapy.CONCLUSION ARHGAP12 expression is elevated in HCC and TKI-resistant HCC,and its regulatory role in FA may underlie the TKI-resistant phenotype.
文摘Focal adhesion kinase(FAK)is a 125-kDa non-receptor protein tyrosine.Growth factors or the clustering of integrins facilitate the rapid phosphorylation of FAK at Tyr-397 and this in turn recruits Src-family protein tyrosine kinases,resulting in the phosphorylation of Tyr-576 and Tyr-577 in the FAK activation loop and full catalytic FAK activation.FAK plays a critical role in the biological processes of normal and cancer cells including the gastrointestinal tract.FAK also plays an important role in the restitution,cell survival and apoptosis and carcinogenesis of the gastrointestinal tract.FAK is over-expressed in cancer cells and its over-expression and elevated activities are associated with motility and invasion of cancer cells.FAK has been proposed as a potential target in cancer therapy.Small molecule inhibitors effectively inhibit the kinase activity of FAK and show a potent inhibitory effect for the proliferation and migration of tumor cells,indicating a high potential for application in cancer therapy.
文摘Focal adhesion kinase(FAK) is a major integrin- dep-endent tyrosine phosphorylated protein, recently, FAK association with colorectal cancer(CRC) has gained at-tention. The various cancer-promoting mechanisms that associated with FAK can be implicated in the progression of CRC. The interactions between structural features of FAK and various kinases could be closely related to growth, survival, and metastasis in CRC cells. These interactions include human epithelial growth factor re-ceptor, c-Met, platelet-derived growth factor receptor, vascular endothelial growth factor receptor, and Src. Such interactions can trigger the survival signaling of CRC cells and are also involved signaling downstream of phosphatidylinositol 3-kinase, AKT, and the extracellular regulated kinase. Based on this scientific background, many pharmaceutical companies are taking efforts to develop FAK inhibitors to treat solid cancer including CRC. Although the anti-cancer efficacies have been noted in many studies, the commercial drugs have not been deve-loped yet. Therefore, the FAK research on CRC is expec-ted to gain momentum and be highly appreciated as a potential field for developing the new drugs. Therefore, the studies on FAK that effect on the progression of human CRC s would be possible to suggest various app-roaches to CRC treatment, and FAK could be a potential target as an anticancer candidate for CRC therapies.
文摘动脉粥样硬化（AS）是一类主要累及大中动脉血管壁的慢性炎症疾病,主要由血脂紊乱引起,炎症反应参与AS发生、发展的全过程.在AS发生发展过程中,炎症细胞、血管内皮细胞、平滑肌细胞和细胞因子、趋化蛋白、黏附分子等相互作用,相互影响动脉粥样化过程.促炎细胞因子可改变动脉粥样硬化早期血管内皮功能,诱导血管内皮细胞表达趋化因子和黏附分子,促进白细胞、淋巴细胞和单核细胞迁移、募集、黏附到发炎的血管壁中.白细胞在动脉血管内膜中被局部产生的细胞因子永久激活,其可以通过刺激清道夫受体的表达和增强细胞介导的氧化来加速巨噬细胞向泡沫细胞的转化,加剧动脉粥样硬化病变进展.致炎细胞因子发挥生物学功能是通过与细胞膜表面受体相互作用,经过跨膜信号转导活化细胞内的相关信号通路,最终促进靶基因的表达.目前认为,丝裂原活化蛋白激酶（mitogen activated protein kinase,MAPKs）、Janus激酶-信号转导子及转录激活子（Janus kinase sig-nal transduction and activator of transcription,JAK-STAT）和核因子kB（nuclear factor kB,NF-kB）是细胞内3条重要信号通路,在炎症信号转导调控中起重要作用.MAPKs是细胞外刺激信号转导至细胞内及核内并引起生物化学功能如细胞增殖、分化、转化及凋亡等的主要信号转导通路.
基金Supported by Grants from the National Natural Science Foundation of China,No 30872513Hebei Provincial Natural Science Foundation of China,No C2008001133 and No C2010000565
文摘AIM:To investigate the effects and mechanism of disruption of focal adhesion kinase(FAK) expression on collagen metabolism in rat hepatic stellate cells(HSC).METHODS:The plasmids expressing FAK short hairpin RNA(shRNA) were transfected into HSC-T6 cells,and the level of FAK expression was determined by both real-time quantitative polymerase chain reaction(QPCR) and Western blotting analysis.The production of type collagen and type collagen in FAK-disrupted cells was analyzed by real-time Q-PCR.The level of collagen metabolism proteins,including matrix metalloproteinases-13(MMP-13) and tissue inhibitors of metalloproteinases-1(TIMP-1) was also determined by both real-time Q-PCR and Western blotting analysis.RESULTS:The transfection of FAK shRNA plasmids into HSC resulted in disrupted FAK expression.Compared with the HK group,the levels of type collagen and type collagen mRNA transcripts in FAK shRNA plas-mid group were signif icantly decreased(0.69 ± 0.03 vs 1.96 ± 0.15,P = 0.000;0.59 ± 0.07 vs 1.62 ± 0.12,P = 0.020).The production of TIMP-1 in this cell type was also signif icantly reduced at both mRNA and protein levels(0.49 ± 0.02 vs 1.72 ± 0.10,P = 0.005;0.76 ± 0.08 vs 2.31 ± 0.24,P = 0.000).However,the expression of MMP-13 mRNA could be significantly up-regulated by the transfection of FAK shRNA plasmids into HSC(1.74 ± 0.20 vs 1.09 ± 0.09,P = 0.000).CONCLUSION:These data support the hypothesis that shRNA-mediated disruption of FAK expression could attenuate extracellular matrix(ECM) synthesis and promote ECM degradation,making FAK a potential target for novel anti-f ibrosis therapies.
文摘目的:本研究旨在证实朝鲜白头翁、人参和甘草的复合水提物(water extract ofPulsatillakoreana(Yabe ex Nakai)Nakai ex T.Mori.,PanaxginsengC.A.Meyer andGlycyrrhizauralensisFisch,WEPPG)的抗血管生成作用。方法:使用纤维母细胞生长因子致血管生成的人类脐静脉内皮细胞模型衡量细胞的增殖、黏附及迁移,同时进行细管形成实验及纤维母细胞生长因子致鸡胚绒毛尿囊膜血管生成实验检测WEPPG的抗血管生成作用。结果:WEPPG能够显著抑制纤维母细胞生长因子所致血管生成的人类脐静脉内皮细胞的增殖、黏附及迁移。信号蛋白分析显示多种蛋白表达变化,如细胞周期素A、p63、KIP2的上调及nibrin蛋白和黏着斑激酶的下调。与对照组相比,WEPPG显著减少了鸡胚绒毛尿囊膜血管生成。结论:本研究的结果证实了WEPPG的抗血管生成作用,这可能是这种药物具有抗癌功效的原因之一。
