AIM: To investigate the expressions of omithine decarboxylase (ODC), MMP-2, and Erk, and their relationship in human colon tumors.METHODS: ODC activity, MMP-2 expression, and mitogenactivated protein (MAP) kinase acti...AIM: To investigate the expressions of omithine decarboxylase (ODC), MMP-2, and Erk, and their relationship in human colon tumors.METHODS: ODC activity, MMP-2 expression, and mitogenactivated protein (MAP) kinase activity (Erk phosphorylation) were determined in 58 surgically removed human colon tumors and their adjacent normal tissues, using [1-14C]-ornithine as a substrate, ELISA assay, and Western blotting, respectively.RESULTS: ODC activity, MMP-2 expression, and Erk phosphorylation were significantly elevated in colon tumors, compared to those in adjacent normal tissues. A significant correlation was observed between ODC activities and MMP-2 levels.CONCLUSION: This is the first report showing a significant correlation between ODC activities and MMP-2 levels in human colon tumors. As MMP-2 is involved in cancer invasion and metastasis, and colon cancer overexpresses ODC, suppression of ODC expression may be a rational approach to treat colon cancer which overexpresses ODC.展开更多
目的探讨加味丹参饮通过调节鸟氨酸脱羧酶(ODC)活性在抗SD大鼠心肌缺血再灌注损伤(IRI)中的保护机制。方法建立SD大鼠心肌IRI模型,设对照组、假手术组、缺血再灌注损伤(IRI)组、加味丹参饮(JWDSY)+IRI组共4组。酶联免疫(ELISA)法测定OD...目的探讨加味丹参饮通过调节鸟氨酸脱羧酶(ODC)活性在抗SD大鼠心肌缺血再灌注损伤(IRI)中的保护机制。方法建立SD大鼠心肌IRI模型,设对照组、假手术组、缺血再灌注损伤(IRI)组、加味丹参饮(JWDSY)+IRI组共4组。酶联免疫(ELISA)法测定ODC含量,实时荧光定量PCR(RT-PCR)方法检测ODC m RNA表达水平,免疫印迹(Western blot)法检测ODC蛋白表达水平,高效液相色谱检测心肌组织中多胺的含量。结果与对照组比较,假手术组的ODC含量、ODC m RNA及蛋白表达水平均无统计学意义(P>0.05);与假手术组比较,IRI组的ODC含量显著增加、ODC m RNA及蛋白表达水平显著升高,差异有统计学意义(P<0.01);与IRI组比较,JWDSY+IRI组ODC含量显著减少,ODC m RNA和蛋白表达水平显著降低,差异有统计学意义(P<0.01)。结论加味丹参饮可能通过调节ODC活性,从而发挥对IRI后心肌的保护作用。展开更多
AIM: To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma.METHODS: Total RNA was extracted from colon cancer tissues and amplified by reverse...AIM: To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma.METHODS: Total RNA was extracted from colon cancer tissues and amplified by reverse-transcription PCR with two primers, which span the whole coding region of ODC. The synthesized ODC cDNA was cloned into vector pQE-30 at restriction sites BamH I and Sal I which constituted recombinant expression plasmid pQE30-ODC. The sequence of inserted fragment was confirmed by DNA sequencing,the fusion protein including 6His-tag was facilitated for purification by Ni-NTA chromatographic column.RESULTS: ODC expression vector was constructed and confirmed with restriction enzyme digestion and subsequent DNA sequencing. The DNA sequence matching on NCBI Blast showed 99 % affinity. The vector was transformed into E.coli M15 and expressed. The expressed ODC protein was verified with Western blotting.CONCLUSION: The ODC prokaryote expression vector is constructed and thus greatly facilitates to study the role of ODC in colorectal carcinoma.展开更多
基金Supported by a Grant Under the Ministry of Education, Science,Sports, and Culture, Japan
文摘AIM: To investigate the expressions of omithine decarboxylase (ODC), MMP-2, and Erk, and their relationship in human colon tumors.METHODS: ODC activity, MMP-2 expression, and mitogenactivated protein (MAP) kinase activity (Erk phosphorylation) were determined in 58 surgically removed human colon tumors and their adjacent normal tissues, using [1-14C]-ornithine as a substrate, ELISA assay, and Western blotting, respectively.RESULTS: ODC activity, MMP-2 expression, and Erk phosphorylation were significantly elevated in colon tumors, compared to those in adjacent normal tissues. A significant correlation was observed between ODC activities and MMP-2 levels.CONCLUSION: This is the first report showing a significant correlation between ODC activities and MMP-2 levels in human colon tumors. As MMP-2 is involved in cancer invasion and metastasis, and colon cancer overexpresses ODC, suppression of ODC expression may be a rational approach to treat colon cancer which overexpresses ODC.
文摘目的探讨加味丹参饮通过调节鸟氨酸脱羧酶(ODC)活性在抗SD大鼠心肌缺血再灌注损伤(IRI)中的保护机制。方法建立SD大鼠心肌IRI模型,设对照组、假手术组、缺血再灌注损伤(IRI)组、加味丹参饮(JWDSY)+IRI组共4组。酶联免疫(ELISA)法测定ODC含量,实时荧光定量PCR(RT-PCR)方法检测ODC m RNA表达水平,免疫印迹(Western blot)法检测ODC蛋白表达水平,高效液相色谱检测心肌组织中多胺的含量。结果与对照组比较,假手术组的ODC含量、ODC m RNA及蛋白表达水平均无统计学意义(P>0.05);与假手术组比较,IRI组的ODC含量显著增加、ODC m RNA及蛋白表达水平显著升高,差异有统计学意义(P<0.01);与IRI组比较,JWDSY+IRI组ODC含量显著减少,ODC m RNA和蛋白表达水平显著降低,差异有统计学意义(P<0.01)。结论加味丹参饮可能通过调节ODC活性,从而发挥对IRI后心肌的保护作用。
基金Scientific Research Fund of national Ministry of Health,No.98-1-173
文摘AIM: To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma.METHODS: Total RNA was extracted from colon cancer tissues and amplified by reverse-transcription PCR with two primers, which span the whole coding region of ODC. The synthesized ODC cDNA was cloned into vector pQE-30 at restriction sites BamH I and Sal I which constituted recombinant expression plasmid pQE30-ODC. The sequence of inserted fragment was confirmed by DNA sequencing,the fusion protein including 6His-tag was facilitated for purification by Ni-NTA chromatographic column.RESULTS: ODC expression vector was constructed and confirmed with restriction enzyme digestion and subsequent DNA sequencing. The DNA sequence matching on NCBI Blast showed 99 % affinity. The vector was transformed into E.coli M15 and expressed. The expressed ODC protein was verified with Western blotting.CONCLUSION: The ODC prokaryote expression vector is constructed and thus greatly facilitates to study the role of ODC in colorectal carcinoma.
