Total cellular RNA was extracted from the osteoblast cells of uewborn rats＇ calvarial bones, and the cDNA containing open-reading frame of osteonectin was amplified by reverse transcription polymerase chain reaction ...Total cellular RNA was extracted from the osteoblast cells of uewborn rats＇ calvarial bones, and the cDNA containing open-reading frame of osteonectin was amplified by reverse transcription polymerase chain reaction （ RT- PCR ）. The obtained product was named On. The On fragment was inserted into pBT- T vector. Then, the On was subdoued, in-frame fused to 3＇ -end of the GST gene of the prokaryotic expression vector pGEX-KG, and the resulting recombinant plasmid was transformed into E. coli BL21 （DE3） pLysS competent cells. A 60 kD fusion protein was expressed after IPTG induction. The On fragment was sequenced, and the sequencing result shows that it shares 99.8% homology with the sequence published in GenBank. The On SDSPAGE analysis exhibits that the On was expressed with the GST gene. There is 10% fused protein in the total E. coli proteins, and the fasion protein is a soluble protein. These experimental results imply that On from Wistar rats was cloned suecessfully and expressed efficiently.展开更多
背景与目的:已经发现富含半胱氨酸的酸性分泌蛋白(secreted protein acidic and richin cysteine,SPARC)在多种恶性肿瘤中有不同程度的表达,并且与肿瘤生物学行为、预后相关。本研究通过检测SPARC在临床Ⅱ期舌体鳞癌的表达,分析其表达...背景与目的:已经发现富含半胱氨酸的酸性分泌蛋白(secreted protein acidic and richin cysteine,SPARC)在多种恶性肿瘤中有不同程度的表达,并且与肿瘤生物学行为、预后相关。本研究通过检测SPARC在临床Ⅱ期舌体鳞癌的表达,分析其表达强度与舌癌预后的关系。方法:收集55例中山大学肿瘤防治中心头颈外科1999年1月至2003年12月临床Ⅱ期(T2N0M0)舌体鳞癌术后标本,25例舌炎症旁组织作对照,免疫组化定性检测SPARC的表达。分析其表达强度与舌癌患者生存、复发、隐匿性淋巴结转移的关系。结果:55例舌癌术后标本中SPARC阳性率为49.1%,25例对照舌上皮组织不表达(P<0.001);SPARC阳性患者的5年累积生存率为30.0%,SPARC阴性患者5年累积生存率为85.3%,统计学分析差异有统计学意义(P=0.005);SPARC在隐匿性淋巴结转移阳性与阴性患者表达率分别为86.7%及35.0%,差异有统计学意义(P=0.001)。SPARC在复发与否患者表达率分别100%及31.7%,差异有统计学意义(P<0.001)。SPARC的表达与隐匿性淋巴结转移、复发呈正相关,相关系数依次为0.460(P<0.001)、0.595(P<0.001)。结论:SPARC在临床Ⅱ期舌体鳞癌中表达高于正常舌上皮组织,其表达与生存率、隐匿性淋巴结转移、复发呈正相关。展开更多
Pancreatic cancer has a considerably poor prognosis with a 5-year survival probability of less than 5% when all stages are combined. Pancreatic cancer is characterized by its dense stroma, which is involved in the cri...Pancreatic cancer has a considerably poor prognosis with a 5-year survival probability of less than 5% when all stages are combined. Pancreatic cancer is characterized by its dense stroma, which is involved in the critical interplay with the tumor cells throughout tumor progression and furthermore, creates a barrier restricting efficient penetration of therapeutics. Alterations in a large number of genes are reflected by a limited number of signaling pathways, which are potential targets. Understanding more about the molecular basis of this devastating cancer type regarding tumor microenvironment, distinct subpopulations of cells, epithelial-to-mesenchymal transition and inflammation will lead to the development of various targeted therapies for controlling tumor growth and metastasis. In this complex scenario of pancreatic cancer, especially members of the “small integrin binding ligand N-linked glycoproteins” (SIBLINGs) and “secreted protein acidic and rich in cysteine” (SPARC) families have emerged due to their prominent roles in properties including proliferation, differentiation, apoptosis, adhesion, migration, angiogenesis, wound repair and regulation of extracellular matrix remodeling. SIBLINGs consist of five members, which include osteopontin (OPN), bone sialoprotein, dentin matrix protein 1, dentin sialophosphoprotein and matrix extracellular phosphoglycoprotein. The SPARC family of modular extracellular proteins is comprised of SPARC/osteonectin (ON) and SPARC-like 1 (hevin); secreted modular calcium binding proteins; testicans and follistatin-like protein. In this review, we especially focus on OPN and ON, elaborating on their special and growing importance in pancreatic cancer diagnosis and prognosis.展开更多
文摘Total cellular RNA was extracted from the osteoblast cells of uewborn rats＇ calvarial bones, and the cDNA containing open-reading frame of osteonectin was amplified by reverse transcription polymerase chain reaction （ RT- PCR ）. The obtained product was named On. The On fragment was inserted into pBT- T vector. Then, the On was subdoued, in-frame fused to 3＇ -end of the GST gene of the prokaryotic expression vector pGEX-KG, and the resulting recombinant plasmid was transformed into E. coli BL21 （DE3） pLysS competent cells. A 60 kD fusion protein was expressed after IPTG induction. The On fragment was sequenced, and the sequencing result shows that it shares 99.8% homology with the sequence published in GenBank. The On SDSPAGE analysis exhibits that the On was expressed with the GST gene. There is 10% fused protein in the total E. coli proteins, and the fasion protein is a soluble protein. These experimental results imply that On from Wistar rats was cloned suecessfully and expressed efficiently.
文摘背景与目的:已经发现富含半胱氨酸的酸性分泌蛋白(secreted protein acidic and richin cysteine,SPARC)在多种恶性肿瘤中有不同程度的表达,并且与肿瘤生物学行为、预后相关。本研究通过检测SPARC在临床Ⅱ期舌体鳞癌的表达,分析其表达强度与舌癌预后的关系。方法:收集55例中山大学肿瘤防治中心头颈外科1999年1月至2003年12月临床Ⅱ期(T2N0M0)舌体鳞癌术后标本,25例舌炎症旁组织作对照,免疫组化定性检测SPARC的表达。分析其表达强度与舌癌患者生存、复发、隐匿性淋巴结转移的关系。结果:55例舌癌术后标本中SPARC阳性率为49.1%,25例对照舌上皮组织不表达(P<0.001);SPARC阳性患者的5年累积生存率为30.0%,SPARC阴性患者5年累积生存率为85.3%,统计学分析差异有统计学意义(P=0.005);SPARC在隐匿性淋巴结转移阳性与阴性患者表达率分别为86.7%及35.0%,差异有统计学意义(P=0.001)。SPARC在复发与否患者表达率分别100%及31.7%,差异有统计学意义(P<0.001)。SPARC的表达与隐匿性淋巴结转移、复发呈正相关,相关系数依次为0.460(P<0.001)、0.595(P<0.001)。结论:SPARC在临床Ⅱ期舌体鳞癌中表达高于正常舌上皮组织,其表达与生存率、隐匿性淋巴结转移、复发呈正相关。
文摘Pancreatic cancer has a considerably poor prognosis with a 5-year survival probability of less than 5% when all stages are combined. Pancreatic cancer is characterized by its dense stroma, which is involved in the critical interplay with the tumor cells throughout tumor progression and furthermore, creates a barrier restricting efficient penetration of therapeutics. Alterations in a large number of genes are reflected by a limited number of signaling pathways, which are potential targets. Understanding more about the molecular basis of this devastating cancer type regarding tumor microenvironment, distinct subpopulations of cells, epithelial-to-mesenchymal transition and inflammation will lead to the development of various targeted therapies for controlling tumor growth and metastasis. In this complex scenario of pancreatic cancer, especially members of the “small integrin binding ligand N-linked glycoproteins” (SIBLINGs) and “secreted protein acidic and rich in cysteine” (SPARC) families have emerged due to their prominent roles in properties including proliferation, differentiation, apoptosis, adhesion, migration, angiogenesis, wound repair and regulation of extracellular matrix remodeling. SIBLINGs consist of five members, which include osteopontin (OPN), bone sialoprotein, dentin matrix protein 1, dentin sialophosphoprotein and matrix extracellular phosphoglycoprotein. The SPARC family of modular extracellular proteins is comprised of SPARC/osteonectin (ON) and SPARC-like 1 (hevin); secreted modular calcium binding proteins; testicans and follistatin-like protein. In this review, we especially focus on OPN and ON, elaborating on their special and growing importance in pancreatic cancer diagnosis and prognosis.
