The use of anti-beta 1 integrin monoclonal antibody in lung cancer treatment has proven beneficial. Here, we developed a novel monoclonal antibody （mAb）, called P5, by immunizing mice with human peripheral blood mon...The use of anti-beta 1 integrin monoclonal antibody in lung cancer treatment has proven beneficial. Here, we developed a novel monoclonal antibody （mAb）, called P5, by immunizing mice with human peripheral blood mononuclear cells （PBMC）. Its anti-tumor effect is now being tested, in a clinical phase Ⅲ trial, in combinato- rial treatments with various chemical drugs. To confirm that P5 indeed binds to beta 1 integrin, cell lysates were immunoprecipitated with commercial anti-beta 1 integrin mAb （TS2/16） and immunoblotted against P5 to reveal a 140 kDa molecular weight band, as expected. Immunoprecipitation with P5 followed by LC/MS protein sequence analysis further verified P5 antigen to be beta 1 integrin. Cisplatin treatment upregulated cell surface expression of beta 1 integrin in A549 cells, while causing inhibition of cell growth. When cells were co-treated with different concentrations of P5 mAb, the cisplatin-mediated inhibitory effect was enhanced in a dose-dependent manner. Our findings show that a combinatorial treatment of P5 mAb and cisplatin in A549 cells resulted in a 30% increase in apoptosis, compared to baseline, and significantly more when compared to either the cisplatin or P5 alone group. The entire peptide sequences in CDR from variable region of Ig heavy and light chain gene for P5 mAb are also disclosed. Together, these results provide evidence of the beneficial effect of P5 mAb in combinatorial treatment of human lung adenocarcinoma.展开更多
目的探讨预防性使用硫酸右旋糖苷(DS)和治疗性使用DS对人胃癌细胞腹腔种植的作用。方法培养胃癌BGC-823细胞,构建裸鼠动物模型:选用BALB/C裸鼠共110只,随机分为预防组(60只)和治疗组(50只)。预防组在注射肿瘤细胞的同时注射DS,治疗组注...目的探讨预防性使用硫酸右旋糖苷(DS)和治疗性使用DS对人胃癌细胞腹腔种植的作用。方法培养胃癌BGC-823细胞,构建裸鼠动物模型:选用BALB/C裸鼠共110只,随机分为预防组(60只)和治疗组(50只)。预防组在注射肿瘤细胞的同时注射DS,治疗组注射肿瘤细胞24 h后注射DS。打开裸鼠腹腔,观察胃癌细胞腹腔种植的数量,所有结节经HE染色证实。应用免疫组织化学染色(SP法)检测大网膜整合素β1蛋白的表达,应用RT-PCR方法检测大网膜整合素β1 m RNA的表达。结果 HE染色结果显示治疗组裸鼠腹腔转移的肿瘤结节数量明显多于预防组,差异有统计学意义(P<0.05),SP法结果显示治疗组整合素β1蛋白的表达量明显高于预防组,差异有统计学意义(P<0.01),RT-PCR结果显示治疗组整合素β1 m RNA的表达量明显高于预防组,差异有统计学意义(P<0.01)。结论 DS预防胃癌腹腔种植转移的效果比治疗胃癌腹腔种植转移的效果更佳。展开更多
目的 利用串联质谱标签(tandem mass tags,TMT)定量蛋白质组学技术研究痹祺胶囊治疗类风湿性关节炎(rheumatoid arthritis,RA)的作用机制。方法 建立II型胶原蛋白诱导的RA大鼠模型,并给予痹祺胶囊进行干预,取对照组(C)、模型组(M)、痹...目的 利用串联质谱标签(tandem mass tags,TMT)定量蛋白质组学技术研究痹祺胶囊治疗类风湿性关节炎(rheumatoid arthritis,RA)的作用机制。方法 建立II型胶原蛋白诱导的RA大鼠模型,并给予痹祺胶囊进行干预,取对照组(C)、模型组(M)、痹祺胶囊0.4 g/kg给药组(BQ)的左膝关节组织,采用TMT定量蛋白质组学技术分析鉴定各组大鼠膝关节组织中的蛋白,以表达差异倍数≥1.2或差异倍数≤0.83且P<0.05筛选M vs C、BQ vs M、BQ vs C差异表达蛋白,并分析经痹祺胶囊干预后有回调趋势的差异表达蛋白。最后通过GeneCards、OMIN数据库检索RA疾病靶点,与回调蛋白进行交集分析得到与RA疾病相关的回调蛋白,对其进行基因本体(gene ontology,GO)功能和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,明晰痹祺胶囊对RA的作用机制。结果 经痹祺胶囊干预后有121个差异蛋白具有回调趋势,其中38个与RA疾病相关,如β2整合素(integrin subunit beta 2,ITGB2)、钙调蛋白2(calmodulin 2,CALM2)、钙调磷酸酶B亚基(calcineurin subunit B type 1,PPP3R1)、前列腺素合成酶(prostacyclin synthase,PTGIS)等。生物信息学分析发现这些回调蛋白参与了与RA相关的多条信号通路,如ITGB2参与调节RA及白细胞跨内皮细胞迁移信号通路;CALM2、PPP3R1通过调节钙信号通路,进而调节T细胞、B细胞受体信号通路、T细胞分化及破骨细胞分化等信号通路;PTGIS参与花生四烯酸代谢途径等。结论 痹祺胶囊可能通过干预ITGB2、PPP3R1、CALM2、PTGIS等蛋白的表达,调节了与RA相关的多条信号通路,发挥免疫抑制、抗炎、活血、抑制骨破坏和骨吸收、抑制血管翳生成等多方面药理作用,从而达到治疗RA的效果。展开更多
基金supported in 2015 by a research grant from Chungbuk National University
文摘The use of anti-beta 1 integrin monoclonal antibody in lung cancer treatment has proven beneficial. Here, we developed a novel monoclonal antibody （mAb）, called P5, by immunizing mice with human peripheral blood mononuclear cells （PBMC）. Its anti-tumor effect is now being tested, in a clinical phase Ⅲ trial, in combinato- rial treatments with various chemical drugs. To confirm that P5 indeed binds to beta 1 integrin, cell lysates were immunoprecipitated with commercial anti-beta 1 integrin mAb （TS2/16） and immunoblotted against P5 to reveal a 140 kDa molecular weight band, as expected. Immunoprecipitation with P5 followed by LC/MS protein sequence analysis further verified P5 antigen to be beta 1 integrin. Cisplatin treatment upregulated cell surface expression of beta 1 integrin in A549 cells, while causing inhibition of cell growth. When cells were co-treated with different concentrations of P5 mAb, the cisplatin-mediated inhibitory effect was enhanced in a dose-dependent manner. Our findings show that a combinatorial treatment of P5 mAb and cisplatin in A549 cells resulted in a 30% increase in apoptosis, compared to baseline, and significantly more when compared to either the cisplatin or P5 alone group. The entire peptide sequences in CDR from variable region of Ig heavy and light chain gene for P5 mAb are also disclosed. Together, these results provide evidence of the beneficial effect of P5 mAb in combinatorial treatment of human lung adenocarcinoma.
文摘目的探讨地黄对去卵巢骨质疏松大鼠股骨骨密度、腰椎骨组织整合素β1 mRNA表达的影响。方法 72只雌性大鼠,随机分为假手术组、模型组、雌二醇组和地黄小、中、大剂量组。假手术组仅行假手术,其余五组行卵巢切除术。术后1w分别灌胃给予17β-雌二醇和地黄小、中、大剂量,连续给药3个月。测定血钙、磷和碱性磷酸酶(ALP),尿钙(u-Ca)、磷、尿脱氧吡啶酚(D-Pyr)和肌酐(Cr)。之后处死动物,取出右侧股骨,测定骨密度;取出第2腰椎,测定骨钙(b-Ca)、磷(b-P)含量;取出第4腰椎,采用实时荧光逆转录聚合酶链反应测定腰椎骨整合素β1 mRNA表达。结果与假手术组相比,模型组血清ALP、u-Ca、D-Pyr/Cr显著增加,股骨密度、腰椎骨整合素β1 mRNA表达均降低。与模型组比较,地黄中、大剂量组和雌二醇组均可使血清ALP、u-Ca、D-Pyr/Cr排出量降低,股骨密度、腰椎骨整合素β1 m RNA表达均增加。结论地黄能抑制由于去卵巢雌激素缺乏引发的骨转换增强,提高骨密度,促进腰椎骨组织整合素β1 m RNA表达,增强骨质量。
文摘目的探讨胰腺癌患者冷冻消融治疗前后血清中CD44v6及整合素β1表达水平的变化及与预后的相关性。方法 44例胰腺癌患者行冷冻消融治疗术,收集患者治疗前后及20名正常人(对照组)的血清标本,采用酶联免疫吸附试验(ELISA)测定CD44v6及整合素β1在血清中的表达水平。结果与对照组比较,患者术前和术后10 d CD44v6和整合素β1的表达水平均较高(P<0.05),而术后1个月的表达水平与对照组差异无统计学意义(P>0.05);术后患者的CD44v6和整合素β1的表达水平均逐渐下降,术后1个月二者的表达水平均明显低于术前(P<0.05)。CD44v6和整合素β1的表达水平与肿瘤大小、分化、临床分期、淋巴结转移及肝转移均明显相关(P<0.05),而与患者性别、年龄及肿瘤位置均无相关性(P>0.05)。CD44v6和整合素β1低表达患者术后的中位生存期分别为14.5个月和14.2个月,而高表达患者仅为9.5个月和9.8个月,二者间差异均有统计学意义(P<0.05)。结论 CD44v6和整合素β1的血清表达水平与胰腺癌的发生、发展、转移及预后等密切相关。
文摘目的探讨预防性使用硫酸右旋糖苷(DS)和治疗性使用DS对人胃癌细胞腹腔种植的作用。方法培养胃癌BGC-823细胞,构建裸鼠动物模型:选用BALB/C裸鼠共110只,随机分为预防组(60只)和治疗组(50只)。预防组在注射肿瘤细胞的同时注射DS,治疗组注射肿瘤细胞24 h后注射DS。打开裸鼠腹腔,观察胃癌细胞腹腔种植的数量,所有结节经HE染色证实。应用免疫组织化学染色(SP法)检测大网膜整合素β1蛋白的表达,应用RT-PCR方法检测大网膜整合素β1 m RNA的表达。结果 HE染色结果显示治疗组裸鼠腹腔转移的肿瘤结节数量明显多于预防组,差异有统计学意义(P<0.05),SP法结果显示治疗组整合素β1蛋白的表达量明显高于预防组,差异有统计学意义(P<0.01),RT-PCR结果显示治疗组整合素β1 m RNA的表达量明显高于预防组,差异有统计学意义(P<0.01)。结论 DS预防胃癌腹腔种植转移的效果比治疗胃癌腹腔种植转移的效果更佳。
文摘目的 利用串联质谱标签(tandem mass tags,TMT)定量蛋白质组学技术研究痹祺胶囊治疗类风湿性关节炎(rheumatoid arthritis,RA)的作用机制。方法 建立II型胶原蛋白诱导的RA大鼠模型,并给予痹祺胶囊进行干预,取对照组(C)、模型组(M)、痹祺胶囊0.4 g/kg给药组(BQ)的左膝关节组织,采用TMT定量蛋白质组学技术分析鉴定各组大鼠膝关节组织中的蛋白,以表达差异倍数≥1.2或差异倍数≤0.83且P<0.05筛选M vs C、BQ vs M、BQ vs C差异表达蛋白,并分析经痹祺胶囊干预后有回调趋势的差异表达蛋白。最后通过GeneCards、OMIN数据库检索RA疾病靶点,与回调蛋白进行交集分析得到与RA疾病相关的回调蛋白,对其进行基因本体(gene ontology,GO)功能和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,明晰痹祺胶囊对RA的作用机制。结果 经痹祺胶囊干预后有121个差异蛋白具有回调趋势,其中38个与RA疾病相关,如β2整合素(integrin subunit beta 2,ITGB2)、钙调蛋白2(calmodulin 2,CALM2)、钙调磷酸酶B亚基(calcineurin subunit B type 1,PPP3R1)、前列腺素合成酶(prostacyclin synthase,PTGIS)等。生物信息学分析发现这些回调蛋白参与了与RA相关的多条信号通路,如ITGB2参与调节RA及白细胞跨内皮细胞迁移信号通路;CALM2、PPP3R1通过调节钙信号通路,进而调节T细胞、B细胞受体信号通路、T细胞分化及破骨细胞分化等信号通路;PTGIS参与花生四烯酸代谢途径等。结论 痹祺胶囊可能通过干预ITGB2、PPP3R1、CALM2、PTGIS等蛋白的表达,调节了与RA相关的多条信号通路,发挥免疫抑制、抗炎、活血、抑制骨破坏和骨吸收、抑制血管翳生成等多方面药理作用,从而达到治疗RA的效果。
