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
目的探讨在高糖作用下,体外培养的小鼠足细胞凋亡及内质网应激凋亡因子Caspase-12 m RNA和蛋白表达水平的变化及左归降糖益肾方含药血浆的调控作用。方法将体外培养的小鼠足细胞随机分为空白组(A组,25 mmol/L葡萄糖,10%空白血浆)、高糖...目的探讨在高糖作用下,体外培养的小鼠足细胞凋亡及内质网应激凋亡因子Caspase-12 m RNA和蛋白表达水平的变化及左归降糖益肾方含药血浆的调控作用。方法将体外培养的小鼠足细胞随机分为空白组(A组,25 mmol/L葡萄糖,10%空白血浆)、高糖组(B组,200 mmol/L葡萄糖,10%空白血浆)、左归降糖益肾方含药血浆组(C组,200 mmol/L葡萄糖,10%含药血浆),4-苯基丁酸含药血浆组(D组,200 mmol/L葡萄糖,10%含药血浆)。培养48 h后,采用免疫荧光细胞化学法、流式细胞术检测足细胞凋亡情况;Western blot法、RT-PCR法分别检测Caspase-12蛋白或m RNA表达水平的变化。结果与A组比较:在高糖状态下B组足细胞凋亡率、Caspase-12 m RNA和蛋白的表达水平均升高,差异有统计学意义(P<0.05)。与B组比较:C组、D组足细胞凋亡率,Caspase-12 m RNA和蛋白的表达水平均降低,差异均有统计学意义(P<0.05)。与C组比较:D组足细胞凋亡率降低,差异有统计学意义(P<0.05);但Caspase-12 m RNA和蛋白的表达水平,差异均无统计学意义(P>0.05)。结论 Caspase-12表达上调,是足细胞凋亡的分子机制之一;10%左归降糖益肾方含药血浆可能通过降低Caspase-12的表达水平,从而保护足细胞。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.
文摘目的探讨在高糖作用下,体外培养的小鼠足细胞凋亡及内质网应激凋亡因子Caspase-12 m RNA和蛋白表达水平的变化及左归降糖益肾方含药血浆的调控作用。方法将体外培养的小鼠足细胞随机分为空白组(A组,25 mmol/L葡萄糖,10%空白血浆)、高糖组(B组,200 mmol/L葡萄糖,10%空白血浆)、左归降糖益肾方含药血浆组(C组,200 mmol/L葡萄糖,10%含药血浆),4-苯基丁酸含药血浆组(D组,200 mmol/L葡萄糖,10%含药血浆)。培养48 h后,采用免疫荧光细胞化学法、流式细胞术检测足细胞凋亡情况;Western blot法、RT-PCR法分别检测Caspase-12蛋白或m RNA表达水平的变化。结果与A组比较:在高糖状态下B组足细胞凋亡率、Caspase-12 m RNA和蛋白的表达水平均升高,差异有统计学意义(P<0.05)。与B组比较:C组、D组足细胞凋亡率,Caspase-12 m RNA和蛋白的表达水平均降低,差异均有统计学意义(P<0.05)。与C组比较:D组足细胞凋亡率降低,差异有统计学意义(P<0.05);但Caspase-12 m RNA和蛋白的表达水平,差异均无统计学意义(P>0.05)。结论 Caspase-12表达上调,是足细胞凋亡的分子机制之一;10%左归降糖益肾方含药血浆可能通过降低Caspase-12的表达水平,从而保护足细胞。
