目的探讨B族链球菌基因cpsG影响人类宫颈癌细胞蛋白组学。方法通过人工合成的方法将cpsG的读码框序列(ORF)融合HA标签连接到质粒pLVX-mCMV-ZsGreen的限制性内切酶EcoRI和BamHI之间构建稳定表达cpsG人类宫颈癌Hela细胞系。提取rLV组和rLV...目的探讨B族链球菌基因cpsG影响人类宫颈癌细胞蛋白组学。方法通过人工合成的方法将cpsG的读码框序列(ORF)融合HA标签连接到质粒pLVX-mCMV-ZsGreen的限制性内切酶EcoRI和BamHI之间构建稳定表达cpsG人类宫颈癌Hela细胞系。提取rLV组和rLV-cpsG组的总蛋白质,并通过12%SDS-PAGE电泳分析后,进行非标记(Label-free)蛋白质酶解肽段质谱分析。结果经蛋白质酶解肽段质谱分析显示,rLV组鉴定出3552个蛋白,rLV-cpsG组鉴定出3702个蛋白;与rLV组相比,rLV-cpsG组中上调的蛋白为229个,下调的蛋白为225个。GO分析结果显示,细胞组分主要包括膜完整性、细胞核、核糖体;分子功能主要包括蛋白结合、ATP结合、GTP结合、DNA结合、RNA结合;生物过程主要包括翻译、代谢、蛋白磷酸化、跨膜。KEGG通路富集分析显示,Cellular Processes通路主要包括细胞生长与凋亡(179个)、细胞活力(72个)、真核细胞群(140个)、转运和代谢(270个);Environmental Information Processing通路有膜转运(11个)、信号转导(324个);Genetic Information Processing通路有复制与修复(51个)、转录(133个)、翻译(328个);Human Diseases通路有癌症(258个);Metabolism通路有氨基酸代谢(110个)、糖代谢(144个);Organismal Systems通路包括内分泌系统(174个)。蛋白质-蛋白质相互作用结果显示,下调的蛋白互作网络主要包括prot1(MGST1)∶prot2(GCLC)、prot1(MGST1)∶prot2(MYH10)、prot1(MGST1)∶prot2(UGDH)、prot1(MGST1)∶prot2(GCLM)、prot1(PKP2)∶prot2(CDH2)、prot1(PKP2)∶prot2(A8K2T7);上调的蛋白互作网络主要包括prot1(SLC7A2)∶prot2(TXN)、prot1(SLC7A2)∶prot2(KYNU)、prot1(SLC7A2)∶prot2(MCAT)、prot1(SLC7A2)∶prot2(JUNB)、prot1(SLC7A2)∶prot2(BOLA2)、prot1(SLC7A2)∶prot2(FN1)。结论B族链球菌基因cpsG影响了人类宫颈癌细胞的蛋白组学,该研究为明确cpsG在人类宫颈癌细胞中发生作用机制及其临床应用提供了基础资料。展开更多
It was recently reported that arsenic trioxide (As2O3) can induce complete remission in patients with acute promyelocytic leukemia (APL). In this present article, the biological effect of As203 on human cervical cance...It was recently reported that arsenic trioxide (As2O3) can induce complete remission in patients with acute promyelocytic leukemia (APL). In this present article, the biological effect of As203 on human cervical cancer HeLa cells and HeLa cells overexpressing Bcl-2 is studied. By MTT and colony forming ability assays, morphology alteration, flow cytometric analysis, DNA gel electrophoresis and in situ cell death detection (TUNEL) , it was found that As2O3 inhibited the growth of HeLa cells and induced G2/M arrest and apoptosis of the cells. RT-PCR, Northern blot, Western blot analysis revealed that As2O3 induced HeLa cell apoptosis possibly via decreasing the expression of c-myc and viral genes. HeLa cells overexpressing Bcl-2 partly resist As2O3 induced apoptosis, which might be relative to preventing the cells from As2O3 caused G2/M block, downregulation of c-myc gene expression and inhibition of viral gene expression was also noted. However, it was found that As2O3 at a high concentration could also induce apoptosis of HeLa cells over-expressing Bcl-2 possibly mainly via downregulating Bcl-2 expression and slightly inhibiting viral gene expression.展开更多
文摘目的探讨B族链球菌基因cpsG影响人类宫颈癌细胞蛋白组学。方法通过人工合成的方法将cpsG的读码框序列(ORF)融合HA标签连接到质粒pLVX-mCMV-ZsGreen的限制性内切酶EcoRI和BamHI之间构建稳定表达cpsG人类宫颈癌Hela细胞系。提取rLV组和rLV-cpsG组的总蛋白质,并通过12%SDS-PAGE电泳分析后,进行非标记(Label-free)蛋白质酶解肽段质谱分析。结果经蛋白质酶解肽段质谱分析显示,rLV组鉴定出3552个蛋白,rLV-cpsG组鉴定出3702个蛋白;与rLV组相比,rLV-cpsG组中上调的蛋白为229个,下调的蛋白为225个。GO分析结果显示,细胞组分主要包括膜完整性、细胞核、核糖体;分子功能主要包括蛋白结合、ATP结合、GTP结合、DNA结合、RNA结合;生物过程主要包括翻译、代谢、蛋白磷酸化、跨膜。KEGG通路富集分析显示,Cellular Processes通路主要包括细胞生长与凋亡(179个)、细胞活力(72个)、真核细胞群(140个)、转运和代谢(270个);Environmental Information Processing通路有膜转运(11个)、信号转导(324个);Genetic Information Processing通路有复制与修复(51个)、转录(133个)、翻译(328个);Human Diseases通路有癌症(258个);Metabolism通路有氨基酸代谢(110个)、糖代谢(144个);Organismal Systems通路包括内分泌系统(174个)。蛋白质-蛋白质相互作用结果显示,下调的蛋白互作网络主要包括prot1(MGST1)∶prot2(GCLC)、prot1(MGST1)∶prot2(MYH10)、prot1(MGST1)∶prot2(UGDH)、prot1(MGST1)∶prot2(GCLM)、prot1(PKP2)∶prot2(CDH2)、prot1(PKP2)∶prot2(A8K2T7);上调的蛋白互作网络主要包括prot1(SLC7A2)∶prot2(TXN)、prot1(SLC7A2)∶prot2(KYNU)、prot1(SLC7A2)∶prot2(MCAT)、prot1(SLC7A2)∶prot2(JUNB)、prot1(SLC7A2)∶prot2(BOLA2)、prot1(SLC7A2)∶prot2(FN1)。结论B族链球菌基因cpsG影响了人类宫颈癌细胞的蛋白组学,该研究为明确cpsG在人类宫颈癌细胞中发生作用机制及其临床应用提供了基础资料。
文摘It was recently reported that arsenic trioxide (As2O3) can induce complete remission in patients with acute promyelocytic leukemia (APL). In this present article, the biological effect of As203 on human cervical cancer HeLa cells and HeLa cells overexpressing Bcl-2 is studied. By MTT and colony forming ability assays, morphology alteration, flow cytometric analysis, DNA gel electrophoresis and in situ cell death detection (TUNEL) , it was found that As2O3 inhibited the growth of HeLa cells and induced G2/M arrest and apoptosis of the cells. RT-PCR, Northern blot, Western blot analysis revealed that As2O3 induced HeLa cell apoptosis possibly via decreasing the expression of c-myc and viral genes. HeLa cells overexpressing Bcl-2 partly resist As2O3 induced apoptosis, which might be relative to preventing the cells from As2O3 caused G2/M block, downregulation of c-myc gene expression and inhibition of viral gene expression was also noted. However, it was found that As2O3 at a high concentration could also induce apoptosis of HeLa cells over-expressing Bcl-2 possibly mainly via downregulating Bcl-2 expression and slightly inhibiting viral gene expression.
