The various fibroproliferative disorders affecting humans have in common excess fibroblast activity and persistent overexpression or dysregulated activity of transforming growth factor beta (TGF-β). Cancer has many s...The various fibroproliferative disorders affecting humans have in common excess fibroblast activity and persistent overexpression or dysregulated activity of transforming growth factor beta (TGF-β). Cancer has many similar characteristics. Antineoplastic drugs can downregulate fibroblast activity and cytokine growth factors. This study evaluates the effect of six antineoplastic drugs on keloid and Dupuytren’s disease fibroblasts. Keloid, normal scar, Dupuytren’s affected palmar fascia, and normal palmar fascia fibroblasts were grown and seeded into Fibroblast Populated Collagen Lattices (FPCLs). The FPCLs were treated with one of six antineoplastic drugs or left untreated as controls. At 7 days, supernatants were extracted from all FPCLs and assayed for expression of Transforming Growth Factor beta (TGF)-β<sub>1</sub> and TGF-β<sub>2</sub>. All six antineoplastic drugs significantly inhibited FPCL contraction in both fibroproliferative conditions compared with the untreated controls (p β<sub>1</sub> and TGF-β<sub>2</sub> expression was downregulated in the supernatants of all FPCLs by the drug exposure. Cytotoxicity did not occur in these studies and was not the reason for the results. Although antineoplastic drugs can have significant side effects when given systemically, these results may be minimized when given to small areas involved in fibroproliferative scarring or when given topically or intralesionally. These in vitro results suggest that antineoplastic drugs may have a utility for treating various fibroproliferative disorders and warrant further investigation.展开更多
背景:增生性瘢痕是以成纤维细胞过度增殖、表皮增厚和角质层功能不良为特征的皮肤纤维化疾病,目前其具体发病机制仍不清楚。目的:基于生物信息学筛选增生性瘢痕相关数据集的核心(Hub)基因及重要信号通路,再用细胞实验加以验证,预测对其...背景:增生性瘢痕是以成纤维细胞过度增殖、表皮增厚和角质层功能不良为特征的皮肤纤维化疾病,目前其具体发病机制仍不清楚。目的:基于生物信息学筛选增生性瘢痕相关数据集的核心(Hub)基因及重要信号通路,再用细胞实验加以验证,预测对其可能有治疗作用的小分子药物。方法:从基因表达综合数据库搜索增生性瘢痕相关的数据集,通过R软件筛选差异表达基因,对差异表达基因进行基因本体论和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Gnomes,KEGG)富集分析,使用String在线平台构建差异表达基因的蛋白质相互作用网络,然后分别利用Cytoscape软件中的Cytohubba和MCODE插件筛选出蛋白质相互作用网络中的关键基因和核心模块,进一步将上述关键基因和构成核心模块的基因求交集得到Hub基因,通过荧光定量PCR验证Hub基因mRNA在人增生性瘢痕与正常皮肤表皮干细胞中的表达差异,并利用人类蛋白图谱中组织学数据验证Hub基因编码蛋白在2种组织中表达量和分布的差异,最后用connectivity map数据库预测针对增生性瘢痕的潜在作用药物。结果与结论:①筛选出的差异表达基因中上调基因102个、下调基因702个,基因本体论和KEGG分析结果显示,富集的信号通路及生物学过程主要涉及紧密连接、花生四烯酸代谢、细胞外基质受体交互、表皮发育和角质化等;②取交集得到8个Hub基因与调控胆固醇代谢的甲羟戊酸途径密切相关,分别是HMGCS1、DHCR7、MSMO1、FDPS、MVK、HMGCR、MVD和ACAT2;③荧光定量PCR结果显示,相比正常皮肤组,增生性瘢痕组HMGCS1、DHCR7、MSMO1、FDPS、HMGCR、MVD和ACAT2 mRNA的表达均显著下降(P<0.05),而MVK mRNA的表达无明显变化(P>0.05);④除MVK外,其余Hub基因编码蛋白在正常皮肤组织中表达水平均高于增生性瘢痕组织(P<0.05);⑤评分排列前10的候选药物包括蛋白激酶A抑制剂(H-89)、丝氨酸蛋白酶抑制剂(Dabigatran-Etexilate)、FLT3抑制剂(舒尼替尼)等,其中白藜芦醇和β-谷甾醇均为植物来源;⑥提示与甲羟戊酸代谢途径密切相关的Hub基因可能通过调控脂质代谢影响表皮结构与功能,这可能是增生性瘢痕的重要发病机制之一,此次研究筛选的小分子化合物可作为治疗增生性瘢痕的候选药物。展开更多
Background:To investigate the role of fibroblast growth factor 2(FGF2)in chemotherapy resistance of colon cancer.Methods:An HCT116/5-fluorouracil(5-FU)-resistant cell line was established,and FGF2 levels were detected...Background:To investigate the role of fibroblast growth factor 2(FGF2)in chemotherapy resistance of colon cancer.Methods:An HCT116/5-fluorouracil(5-FU)-resistant cell line was established,and FGF2 levels were detected in a sensitive cell group(HCT116)and a resistant cell group(HCT1116-R)using different methods.Fibroblast growth factor 2 levels in the medium were determined by enzyme-linked immunoassay.The protein expressions of FGF2,fibroblast growth factor receptor 1(FGFR1),and phospho-FGFR1 were assessed by Western blotting,and FGF2 mRNA levels were detected by quantitative real-time polymerase chain reaction.Fibroblast growth factor 2 recombinant protein was added to sensitive cells,and FGFR inhibitor AZD4547 was added to resistant cells,and the cell survival rate was determined using the cell counting kit-8 method and the protein expressions of PI3K(phosphatidylinositol 3 kinase),p-PI3K(phospho-PI3K),Akt(protein kinase B),p-Akt(phospho-Akt),mammalian target of rapamycin(mTOR),p-mTOR(phospho-mTOR),Bad(Bcl-xL/Bcl-2-associated death promoter),NF-κB(nuclear factorκB),GSK-3(glycogen synthase kinase-3),FKHR(forkhead box protein O1),and PTEN(phosphatase and tensin homolog deleted on chromosome ten)were detected by Western blotting.Results:Fibroblast growth factor 2 protein and mRNA expression levels in the HCT116-R group were significantly higher than those in the HCT116 group.Fibroblast growth factor 2 increased the survival rate of HCT116 cells;improved tolerance to 5-FU;upregulated p-PI3K,p-Akt,and p-mTOR;and downregulated Bad.The FGFR inhibitor AZD4547 decreased cell survival rate and tolerance to 5-FU;downregulated p-PI3K,p-Akt,and p-mTOR expression;and upregulated Bad.Conclusions:Fibroblast growth factor 2 promotes chemotherapy tolerance in colon cancer cells by activating the Akt/mTOR and Akt/Bad signaling pathways downstream of PI3K.展开更多
文摘The various fibroproliferative disorders affecting humans have in common excess fibroblast activity and persistent overexpression or dysregulated activity of transforming growth factor beta (TGF-β). Cancer has many similar characteristics. Antineoplastic drugs can downregulate fibroblast activity and cytokine growth factors. This study evaluates the effect of six antineoplastic drugs on keloid and Dupuytren’s disease fibroblasts. Keloid, normal scar, Dupuytren’s affected palmar fascia, and normal palmar fascia fibroblasts were grown and seeded into Fibroblast Populated Collagen Lattices (FPCLs). The FPCLs were treated with one of six antineoplastic drugs or left untreated as controls. At 7 days, supernatants were extracted from all FPCLs and assayed for expression of Transforming Growth Factor beta (TGF)-β<sub>1</sub> and TGF-β<sub>2</sub>. All six antineoplastic drugs significantly inhibited FPCL contraction in both fibroproliferative conditions compared with the untreated controls (p β<sub>1</sub> and TGF-β<sub>2</sub> expression was downregulated in the supernatants of all FPCLs by the drug exposure. Cytotoxicity did not occur in these studies and was not the reason for the results. Although antineoplastic drugs can have significant side effects when given systemically, these results may be minimized when given to small areas involved in fibroproliferative scarring or when given topically or intralesionally. These in vitro results suggest that antineoplastic drugs may have a utility for treating various fibroproliferative disorders and warrant further investigation.
文摘背景:增生性瘢痕是以成纤维细胞过度增殖、表皮增厚和角质层功能不良为特征的皮肤纤维化疾病,目前其具体发病机制仍不清楚。目的:基于生物信息学筛选增生性瘢痕相关数据集的核心(Hub)基因及重要信号通路,再用细胞实验加以验证,预测对其可能有治疗作用的小分子药物。方法:从基因表达综合数据库搜索增生性瘢痕相关的数据集,通过R软件筛选差异表达基因,对差异表达基因进行基因本体论和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Gnomes,KEGG)富集分析,使用String在线平台构建差异表达基因的蛋白质相互作用网络,然后分别利用Cytoscape软件中的Cytohubba和MCODE插件筛选出蛋白质相互作用网络中的关键基因和核心模块,进一步将上述关键基因和构成核心模块的基因求交集得到Hub基因,通过荧光定量PCR验证Hub基因mRNA在人增生性瘢痕与正常皮肤表皮干细胞中的表达差异,并利用人类蛋白图谱中组织学数据验证Hub基因编码蛋白在2种组织中表达量和分布的差异,最后用connectivity map数据库预测针对增生性瘢痕的潜在作用药物。结果与结论:①筛选出的差异表达基因中上调基因102个、下调基因702个,基因本体论和KEGG分析结果显示,富集的信号通路及生物学过程主要涉及紧密连接、花生四烯酸代谢、细胞外基质受体交互、表皮发育和角质化等;②取交集得到8个Hub基因与调控胆固醇代谢的甲羟戊酸途径密切相关,分别是HMGCS1、DHCR7、MSMO1、FDPS、MVK、HMGCR、MVD和ACAT2;③荧光定量PCR结果显示,相比正常皮肤组,增生性瘢痕组HMGCS1、DHCR7、MSMO1、FDPS、HMGCR、MVD和ACAT2 mRNA的表达均显著下降(P<0.05),而MVK mRNA的表达无明显变化(P>0.05);④除MVK外,其余Hub基因编码蛋白在正常皮肤组织中表达水平均高于增生性瘢痕组织(P<0.05);⑤评分排列前10的候选药物包括蛋白激酶A抑制剂(H-89)、丝氨酸蛋白酶抑制剂(Dabigatran-Etexilate)、FLT3抑制剂(舒尼替尼)等,其中白藜芦醇和β-谷甾醇均为植物来源;⑥提示与甲羟戊酸代谢途径密切相关的Hub基因可能通过调控脂质代谢影响表皮结构与功能,这可能是增生性瘢痕的重要发病机制之一,此次研究筛选的小分子化合物可作为治疗增生性瘢痕的候选药物。
基金supported by the National Natural Science Foundation of China (no. 81904109)the Natural Science Project of Hunan Provincial Department of Science and Technology (no.2023JJ30361, no. 2019JJ50344).
文摘Background:To investigate the role of fibroblast growth factor 2(FGF2)in chemotherapy resistance of colon cancer.Methods:An HCT116/5-fluorouracil(5-FU)-resistant cell line was established,and FGF2 levels were detected in a sensitive cell group(HCT116)and a resistant cell group(HCT1116-R)using different methods.Fibroblast growth factor 2 levels in the medium were determined by enzyme-linked immunoassay.The protein expressions of FGF2,fibroblast growth factor receptor 1(FGFR1),and phospho-FGFR1 were assessed by Western blotting,and FGF2 mRNA levels were detected by quantitative real-time polymerase chain reaction.Fibroblast growth factor 2 recombinant protein was added to sensitive cells,and FGFR inhibitor AZD4547 was added to resistant cells,and the cell survival rate was determined using the cell counting kit-8 method and the protein expressions of PI3K(phosphatidylinositol 3 kinase),p-PI3K(phospho-PI3K),Akt(protein kinase B),p-Akt(phospho-Akt),mammalian target of rapamycin(mTOR),p-mTOR(phospho-mTOR),Bad(Bcl-xL/Bcl-2-associated death promoter),NF-κB(nuclear factorκB),GSK-3(glycogen synthase kinase-3),FKHR(forkhead box protein O1),and PTEN(phosphatase and tensin homolog deleted on chromosome ten)were detected by Western blotting.Results:Fibroblast growth factor 2 protein and mRNA expression levels in the HCT116-R group were significantly higher than those in the HCT116 group.Fibroblast growth factor 2 increased the survival rate of HCT116 cells;improved tolerance to 5-FU;upregulated p-PI3K,p-Akt,and p-mTOR;and downregulated Bad.The FGFR inhibitor AZD4547 decreased cell survival rate and tolerance to 5-FU;downregulated p-PI3K,p-Akt,and p-mTOR expression;and upregulated Bad.Conclusions:Fibroblast growth factor 2 promotes chemotherapy tolerance in colon cancer cells by activating the Akt/mTOR and Akt/Bad signaling pathways downstream of PI3K.
