Objective: To characterize the expression of ST13 protein in human tissuesfor investigation of the function of colorectal cancer related gene ST13. Methods: ST13 ORF wascloned and over-expressed in E.coli. The recombi...Objective: To characterize the expression of ST13 protein in human tissuesfor investigation of the function of colorectal cancer related gene ST13. Methods: ST13 ORF wascloned and over-expressed in E.coli. The recombinant ST13 protein was purified by affinitychromatography. ST13 monoclonal antibodies were generated and affinity purified with the recombinantprotein. Immunoblot and immunohistochemical staining were employed to analyze ST13 proteinexpression in human tissues. Results: The expression and purification of the recombinant ST13protein were confirmed by SDS-PAGE. The protein yield reached about 2.5 mg/L of induced bacterialculture with a purity of 91.3%. Three strains of hybridoma were obtained with antibody titers from10~4 to 10~5 in ascites fluids and with high specificity for ST13 protein. Immunoblot showed thatthe apparent Mr of ST13 protein in SW480 cells and human tissues estimated by SDS-PAGE mobility wasapproximately 50 000, which was about 10 000 larger than the 41 324 calculated, but theglycosylation of the protein was excluded. Computer modeling revealed the protein to be ahydrophilic molecule. Immunohistochemical staining showed that ST13 protein was evenly distributedin cytoplasm and expressed in colon, stomach, liver, and other epithelial cells. Differences in thestaining intensity of the protein were observed between normal and cancer tissues as well as amongdifferent normal or carcinoma tissues. Conclusion: ST13 protein is a cytoplasmic molecule with anapparent Mr of 50 000. The protein is expressed in colorectal and other epithelial tissues. Theexpression level of the protein is down-regulated in colorectal cancer and varies among differentnormal and/or carcinoma tissues. Comparison of cDNA sequences and protein characteristics indicatesthat ST13 protein and hsp70-interacting protein (Hip) are same proteins, raising the possibilitythat ST13 protein is involved in the development of colorectal cancer through Hsp70 molecularchaperone machinery.展开更多
目的:测定肿瘤发生抑制基因1 3号(suppression of tumorigenicity 13,ST13)蛋白和热休克蛋白70(heat shock protein 70,HSP70)在胃息肉癌变过程中的表达水平,揭示ST13和HSP70在胃息肉及胃癌形成过程中的的相互关系及临床意义.方法:采用...目的:测定肿瘤发生抑制基因1 3号(suppression of tumorigenicity 13,ST13)蛋白和热休克蛋白70(heat shock protein 70,HSP70)在胃息肉癌变过程中的表达水平,揭示ST13和HSP70在胃息肉及胃癌形成过程中的的相互关系及临床意义.方法:采用免疫组织化学SABC法检测ST13和HSP70在106例胃黏膜组织中的表达水平,按组织病理类型分为:正常胃黏膜组30例、增生性息肉组30例、腺瘤样息肉组16例、胃癌组30例,研究二者在胃息肉癌变过程中表达的相互作用及临床意义.结果:采用Krusral-wallis H方法检验HSP70在正常胃黏膜组、增生性息肉组、腺瘤样息肉组、胃癌组中的表达秩均值分别为32.35、40.47、63.25、82.48,有非常显著的差异性(P<0.01),表达呈逐渐递增趋势.ST13在正常胃黏膜、增生性息肉、腺瘤样息肉、胃癌组织中的表达秩均值分别为68.25、59.77、44.91、37.07,有非常显著差异性(P<0.01),表达呈逐渐递减趋势.HSP70和ST13在腺瘤样息肉组和胃癌组中表达呈非常显著负相关(P<0.01).结论:(1)HSP70蛋白在正常胃黏膜、胃增生性息肉、胃腺瘤样息肉、胃癌组织中的表达水平呈显著增高趋势;(2)ST13蛋白在正常胃黏膜、胃增生性息肉、胃腺瘤样息肉、胃癌组织中的表达水平呈显著降低趋势;(3)在胃腺瘤样息肉及胃癌组织中HSP70蛋白与ST13蛋白可能相互作用,其相互关系为负相关,HSP70蛋白可能对其分子共伴侣ST13蛋白产生直接或间接下调作用.展开更多
This article is to summarize the molecular and functional analysis of the gene “suppression of tumorigenicity 13” (ST13). ST13 is in fact the gene encoding Hsp70 interacting protein (Hip), a co-factor (co-chaperone)...This article is to summarize the molecular and functional analysis of the gene “suppression of tumorigenicity 13” (ST13). ST13 is in fact the gene encoding Hsp70 interacting protein (Hip), a co-factor (co-chaperone) of the 70-kDa heat shock proteins (Hsc/Hsp70). By collaborating with other positive co-factors such as Hsp40 and the Hsp70-Hsp90 organizing protein (Hop), or competing with negative co-factors such as Bcl2-associated athanogen 1 (Bag1), Hip facilitates may facilitate the chaperone function of Hsc/Hsp70 in protein folding and repair, and in controlling the activity of regulatory proteins such as steroid receptors and regulators of proliferation or apoptosis. Although the nomenclature of ST13 implies a role in the suppression of tumorigenicity (ST), to date available experimental data are not sufficient to support its role in cancer development, except for the possible down-regulation of ST13 in gastric and colorectal cancers. Further investigation of this gene at the physiological level would benefit our understanding of diseases such as endocrinological disorders, cancer, and neurodegeneration commonly associated with protein misfolding.展开更多
We identified a novel gene ST13 from a subtraetive eDNA library of normal intestinal mueosa in 1993, more studies showed that ST13 was a co-chaperone of Hsp70s. Recently we detected the ST13 gene expression in tumor t...We identified a novel gene ST13 from a subtraetive eDNA library of normal intestinal mueosa in 1993, more studies showed that ST13 was a co-chaperone of Hsp70s. Recently we detected the ST13 gene expression in tumor tissue and adjacent normal tissue of the same colorectal cancer patient and investigated if the ST13 gene expression might have any prognostic value. Analysis was performed at molecular level by reverse transcription-PCR using real-time detection method. We measured two genes simultaneously, ST13 as the target gene and glyceraldehydes-3-phosphate dehydrogenase as a reference gene, in primary colorectal tumor specimens and tumor-adjacent normal mucosa specimens from 50 colorectal cancer patients. The expression levels of the ST13 gene were significantly decreased in primary tumors compared with adjacent mucosa （P〈0.05）. But there were no significant differences in the expression of ST13 as compared depth, lymph node metastasis and disease-specific survival. with different Dukes＇ stage, tumor differentiation grade, invasion展开更多
文摘Objective: To characterize the expression of ST13 protein in human tissuesfor investigation of the function of colorectal cancer related gene ST13. Methods: ST13 ORF wascloned and over-expressed in E.coli. The recombinant ST13 protein was purified by affinitychromatography. ST13 monoclonal antibodies were generated and affinity purified with the recombinantprotein. Immunoblot and immunohistochemical staining were employed to analyze ST13 proteinexpression in human tissues. Results: The expression and purification of the recombinant ST13protein were confirmed by SDS-PAGE. The protein yield reached about 2.5 mg/L of induced bacterialculture with a purity of 91.3%. Three strains of hybridoma were obtained with antibody titers from10~4 to 10~5 in ascites fluids and with high specificity for ST13 protein. Immunoblot showed thatthe apparent Mr of ST13 protein in SW480 cells and human tissues estimated by SDS-PAGE mobility wasapproximately 50 000, which was about 10 000 larger than the 41 324 calculated, but theglycosylation of the protein was excluded. Computer modeling revealed the protein to be ahydrophilic molecule. Immunohistochemical staining showed that ST13 protein was evenly distributedin cytoplasm and expressed in colon, stomach, liver, and other epithelial cells. Differences in thestaining intensity of the protein were observed between normal and cancer tissues as well as amongdifferent normal or carcinoma tissues. Conclusion: ST13 protein is a cytoplasmic molecule with anapparent Mr of 50 000. The protein is expressed in colorectal and other epithelial tissues. Theexpression level of the protein is down-regulated in colorectal cancer and varies among differentnormal and/or carcinoma tissues. Comparison of cDNA sequences and protein characteristics indicatesthat ST13 protein and hsp70-interacting protein (Hip) are same proteins, raising the possibilitythat ST13 protein is involved in the development of colorectal cancer through Hsp70 molecularchaperone machinery.
文摘目的:测定肿瘤发生抑制基因1 3号(suppression of tumorigenicity 13,ST13)蛋白和热休克蛋白70(heat shock protein 70,HSP70)在胃息肉癌变过程中的表达水平,揭示ST13和HSP70在胃息肉及胃癌形成过程中的的相互关系及临床意义.方法:采用免疫组织化学SABC法检测ST13和HSP70在106例胃黏膜组织中的表达水平,按组织病理类型分为:正常胃黏膜组30例、增生性息肉组30例、腺瘤样息肉组16例、胃癌组30例,研究二者在胃息肉癌变过程中表达的相互作用及临床意义.结果:采用Krusral-wallis H方法检验HSP70在正常胃黏膜组、增生性息肉组、腺瘤样息肉组、胃癌组中的表达秩均值分别为32.35、40.47、63.25、82.48,有非常显著的差异性(P<0.01),表达呈逐渐递增趋势.ST13在正常胃黏膜、增生性息肉、腺瘤样息肉、胃癌组织中的表达秩均值分别为68.25、59.77、44.91、37.07,有非常显著差异性(P<0.01),表达呈逐渐递减趋势.HSP70和ST13在腺瘤样息肉组和胃癌组中表达呈非常显著负相关(P<0.01).结论:(1)HSP70蛋白在正常胃黏膜、胃增生性息肉、胃腺瘤样息肉、胃癌组织中的表达水平呈显著增高趋势;(2)ST13蛋白在正常胃黏膜、胃增生性息肉、胃腺瘤样息肉、胃癌组织中的表达水平呈显著降低趋势;(3)在胃腺瘤样息肉及胃癌组织中HSP70蛋白与ST13蛋白可能相互作用,其相互关系为负相关,HSP70蛋白可能对其分子共伴侣ST13蛋白产生直接或间接下调作用.
基金Project supported by the National Basic Research Program (973) of China (No. 2004CB518707)the National Natureal Science Foundation of China (No. 30400521)
文摘This article is to summarize the molecular and functional analysis of the gene “suppression of tumorigenicity 13” (ST13). ST13 is in fact the gene encoding Hsp70 interacting protein (Hip), a co-factor (co-chaperone) of the 70-kDa heat shock proteins (Hsc/Hsp70). By collaborating with other positive co-factors such as Hsp40 and the Hsp70-Hsp90 organizing protein (Hop), or competing with negative co-factors such as Bcl2-associated athanogen 1 (Bag1), Hip facilitates may facilitate the chaperone function of Hsc/Hsp70 in protein folding and repair, and in controlling the activity of regulatory proteins such as steroid receptors and regulators of proliferation or apoptosis. Although the nomenclature of ST13 implies a role in the suppression of tumorigenicity (ST), to date available experimental data are not sufficient to support its role in cancer development, except for the possible down-regulation of ST13 in gastric and colorectal cancers. Further investigation of this gene at the physiological level would benefit our understanding of diseases such as endocrinological disorders, cancer, and neurodegeneration commonly associated with protein misfolding.
文摘We identified a novel gene ST13 from a subtraetive eDNA library of normal intestinal mueosa in 1993, more studies showed that ST13 was a co-chaperone of Hsp70s. Recently we detected the ST13 gene expression in tumor tissue and adjacent normal tissue of the same colorectal cancer patient and investigated if the ST13 gene expression might have any prognostic value. Analysis was performed at molecular level by reverse transcription-PCR using real-time detection method. We measured two genes simultaneously, ST13 as the target gene and glyceraldehydes-3-phosphate dehydrogenase as a reference gene, in primary colorectal tumor specimens and tumor-adjacent normal mucosa specimens from 50 colorectal cancer patients. The expression levels of the ST13 gene were significantly decreased in primary tumors compared with adjacent mucosa （P〈0.05）. But there were no significant differences in the expression of ST13 as compared depth, lymph node metastasis and disease-specific survival. with different Dukes＇ stage, tumor differentiation grade, invasion
