干旱是制约甘蔗产业发展的重要因素之一。前人研究表明,斑茅具有良好的抗性基因,可以通过远缘杂交遗传给后代。本研究以斑茅与甘蔗杂交F1代无性系YCE96-40为材料,对苗期干旱处理0 h和24 h后的叶和根进行转录组测序分析,比较了根和叶在...干旱是制约甘蔗产业发展的重要因素之一。前人研究表明,斑茅具有良好的抗性基因,可以通过远缘杂交遗传给后代。本研究以斑茅与甘蔗杂交F1代无性系YCE96-40为材料,对苗期干旱处理0 h和24 h后的叶和根进行转录组测序分析,比较了根和叶在转录水平上响应干旱的差异,鉴定出21,885个(DR vs CR:10176,DL vs CL:7907)差异表达基因(DEGs),根中差异表达基因多于叶中,说明根对干旱胁迫响应更为剧烈。GO功能富集分析发现,根和叶中DEGs均富集到与脱水反应相关及激素信号转导过程相关的条目,比如“对渗透胁迫生物过程的响应”和“对缺水生物过程的反应”等。与叶不同的是,根中大量DEGs显著富集到与细胞膜相关的条目。在根中鉴定出多个木质素相关DEGs,表明木质素参与了根的干旱响应。通过对所有DEGs进行WGCNA分析,共鉴定出11个基因共表达模块,其中与干旱处理后的根显著相关有5个模块,与干旱处理后的叶显著相关的有2个模块。进一步筛选出了26个转录因子为甘蔗干旱响应的候选转录因子,构建了调控网络。研究结果为进一步理解甘蔗抗旱性的分子机制及甘蔗抗旱性育种提供了理论指导。展开更多
Objective To investigate the in vivo immunomodulatory and anti-tumor mechanisms of the combined treatment of novel Four-Herb formula(4HF)and doxorubicin in triple-negative breast cancer(TNBC).Methods Murine-derived tr...Objective To investigate the in vivo immunomodulatory and anti-tumor mechanisms of the combined treatment of novel Four-Herb formula(4HF)and doxorubicin in triple-negative breast cancer(TNBC).Methods Murine-derived triple-negative mammary carcinoma cell line,4T1 cells,was cultured and inoculated into mouse mammary glands.Sixty-six mice were randomly assigned into 6 groups(n=11 in ench):naive,control,LD 4HF(low dose 4HF),HD 4HF(high dose 4HF),LD 4HF+D(low dose and doxorubicin),and D(doxorubicin).Apart from the naive group,each mouse received subcutaneous inoculation with 5×10^(5)4T1 cells resuspended in 100µL of normal saline in the mammary fat pads.Starting from the day of tumor cell inoculation,tumors were grown for 6 days.The LD and HD groups received daily oral gavage of 658 and 2,630 mg/kg 4HF,respectively.The LD 4HF+D group received daily oral gavage of 658 mg/kg 4HF and weekly intraperitoneal injection of doxorubicin(5 mg/kg).The D group received weekly intraperitoneal injections of doxorubicin(5 mg/kg).The treatment naive mice received daily oral gavage of 0.2 mL double distilled water and 0.1 mL normal saline via intraperitoneal injection once a week.The control group received daily oral gavage of 0.2 mL double-distilled water.The treatment period was 30 days.At the end of treatment,mice organs were harvested to analyze immunological activities via immunophenotyping,gene and multiplex analysis,histological staining,and gut microbiota analysis.Results Mice treated with the combination of 4HF and doxorubicin resulted in significantly reduced tumor and spleen burdens(P<0.05),altered the hypoxia and overall immune lymphocyte landscape,and manipulated gut microbiota to favor the anti-tumor immunological activities.Moreover,immunosuppressive genes,cytokines,and chemokines such as C-C motif chemokine 2 and interleukin-10 of tumors were significantly downregulated(P<0.05).4HF-doxorubicin combination treatment demonstrated synergetic activities and was most effective in activating the anti-tumor immune response(P<0.05).Conclusion The above results provide evidence for evaluating the immune regulating mechanisms of 4HF in breast cancer and support its clinical significance in its potential as an adjunctive therapeutic agent or immune supplement.展开更多
文摘干旱是制约甘蔗产业发展的重要因素之一。前人研究表明,斑茅具有良好的抗性基因,可以通过远缘杂交遗传给后代。本研究以斑茅与甘蔗杂交F1代无性系YCE96-40为材料,对苗期干旱处理0 h和24 h后的叶和根进行转录组测序分析,比较了根和叶在转录水平上响应干旱的差异,鉴定出21,885个(DR vs CR:10176,DL vs CL:7907)差异表达基因(DEGs),根中差异表达基因多于叶中,说明根对干旱胁迫响应更为剧烈。GO功能富集分析发现,根和叶中DEGs均富集到与脱水反应相关及激素信号转导过程相关的条目,比如“对渗透胁迫生物过程的响应”和“对缺水生物过程的反应”等。与叶不同的是,根中大量DEGs显著富集到与细胞膜相关的条目。在根中鉴定出多个木质素相关DEGs,表明木质素参与了根的干旱响应。通过对所有DEGs进行WGCNA分析,共鉴定出11个基因共表达模块,其中与干旱处理后的根显著相关有5个模块,与干旱处理后的叶显著相关的有2个模块。进一步筛选出了26个转录因子为甘蔗干旱响应的候选转录因子,构建了调控网络。研究结果为进一步理解甘蔗抗旱性的分子机制及甘蔗抗旱性育种提供了理论指导。
基金Supported by the Funding of the State Key Laboratory of Research on Bioactivities and Clinical Applications of Medicinal Plants(The Chinese University of Hong Kong/CUHK)from Innovation and Technology Commission,Hong Kong,Li Dak Sum Yip Yio Chin R&D Centre for Chinese Medicine,CUHK,Hong Kong and C.C.Wu Cultural Foundation。
文摘Objective To investigate the in vivo immunomodulatory and anti-tumor mechanisms of the combined treatment of novel Four-Herb formula(4HF)and doxorubicin in triple-negative breast cancer(TNBC).Methods Murine-derived triple-negative mammary carcinoma cell line,4T1 cells,was cultured and inoculated into mouse mammary glands.Sixty-six mice were randomly assigned into 6 groups(n=11 in ench):naive,control,LD 4HF(low dose 4HF),HD 4HF(high dose 4HF),LD 4HF+D(low dose and doxorubicin),and D(doxorubicin).Apart from the naive group,each mouse received subcutaneous inoculation with 5×10^(5)4T1 cells resuspended in 100µL of normal saline in the mammary fat pads.Starting from the day of tumor cell inoculation,tumors were grown for 6 days.The LD and HD groups received daily oral gavage of 658 and 2,630 mg/kg 4HF,respectively.The LD 4HF+D group received daily oral gavage of 658 mg/kg 4HF and weekly intraperitoneal injection of doxorubicin(5 mg/kg).The D group received weekly intraperitoneal injections of doxorubicin(5 mg/kg).The treatment naive mice received daily oral gavage of 0.2 mL double distilled water and 0.1 mL normal saline via intraperitoneal injection once a week.The control group received daily oral gavage of 0.2 mL double-distilled water.The treatment period was 30 days.At the end of treatment,mice organs were harvested to analyze immunological activities via immunophenotyping,gene and multiplex analysis,histological staining,and gut microbiota analysis.Results Mice treated with the combination of 4HF and doxorubicin resulted in significantly reduced tumor and spleen burdens(P<0.05),altered the hypoxia and overall immune lymphocyte landscape,and manipulated gut microbiota to favor the anti-tumor immunological activities.Moreover,immunosuppressive genes,cytokines,and chemokines such as C-C motif chemokine 2 and interleukin-10 of tumors were significantly downregulated(P<0.05).4HF-doxorubicin combination treatment demonstrated synergetic activities and was most effective in activating the anti-tumor immune response(P<0.05).Conclusion The above results provide evidence for evaluating the immune regulating mechanisms of 4HF in breast cancer and support its clinical significance in its potential as an adjunctive therapeutic agent or immune supplement.
