The aim of this study was to elucidate the expression and regulation of the c.kit protein in spermatogenesis of locusts. Immunohistochemistry and biological statistics were used to investigate the expression of the c-...The aim of this study was to elucidate the expression and regulation of the c.kit protein in spermatogenesis of locusts. Immunohistochemistry and biological statistics were used to investigate the expression of the c-kit protein in four representative phases of spermatogenesis of three dominant species of locusts of Arcypteridae （Orthoptera： Acridoidea）, namely, Omocestus viridulus （Linnaeus）, Euchorthippus unicolor （Ikonn.）, and Euchorthippus vittatus Zheng, and so on, in Siping area of Jilin Province, China. The results revealed the following： （1） There was weak positive expression of the c-kit protein in spermatogonia and the positive granules were thinner; （2） there was a strong positive expression of the c-kit protein in primary spermatocyte and the positive granules became the largest than in all developmental stages; （3） the c-kit protein positive expression became stronger in secondary spermatocyte, while the positive granules became thinner; （4） there was strong positive expression of the c-kit protein and the positive granules were thinner in mature sperm, which were distributed on its head and tail; （5） there were strong positive protein granules massing at the end of spermary; （6） the positive intensity of the c-kit protein in spermatogenesis was significantly different among different species of locusts. The data suggested that the c-kit protein may play a crucial role in spermatogenesis as well as maintain the physiological action of sperms and fertilization, regulate the developmental speed of spermatogenesis, and/or maintain species isolation, etc.展开更多
The tyrosine kinase receptor III, c-Kit/stem cell factor receptor and its ligand, human stem cell factor (huSCF) are the predominant regulator of mitogenesis in the hematopoietic stem and progenitor cells. However, ga...The tyrosine kinase receptor III, c-Kit/stem cell factor receptor and its ligand, human stem cell factor (huSCF) are the predominant regulator of mitogenesis in the hematopoietic stem and progenitor cells. However, gain-of-function mutations alter c-Kit auto-regulatory mechanisms to aberrant c-Kit signaling, leading to the onset or progression of cancerous transformations. The most common mutation of c-Kit is the substitution of aspartic acid residue in position 816 to valine (D816V), which is majorly responsible for its ligand-independent constitutive activation, and is implicated in hematopoietic malignancies. Currently, molecular targeted therapy is increasingly becoming a hot spot due to its specificity and low toxicity. As the molecular mechanisms responsible for D816V-c-Kit mediated tumorogenicity are largely unknown, in this study, we aimed to investigate the D816V-c-Kit signaling mediated downstream molecular targets. Specifically, we created c-Kit active mutant form D816V and performed inducible gene expression of mutant D816V-c-Kit in monomyelocytic cell line U937. Mutant D816V-c-Kit expressing cells revealed significantly enhanced cellular mitogenic activity compared to wild-type c-Kit expressing cells independent of huSCF. To examine the molecular targets regulating tumorogenic proliferation, we evaluated the consequences of mutant D816V-c-Kit expression on downstream gene expression profile by high throughput microarray technology. The levels of some of the relevant genes (PIK3CB, eIF4B, PRKCDBP, MOAP1) were validated by quantitative polymerase chain reaction. SLA, STAT5B, MAP3K2 and MAPK14 emerged as important downstream molecular targets of mutant D816V-c-Kit. Further, by dissecting the signaling pathways, we also demonstrated that the D816V-c-Kit mediated hematopoietic cell proliferation is dependent on molecular target p38 MAP kinase.展开更多
AIM: To investigate the prognostic significance of c-Kitgen emutation and DNA ploidy in gastointestinal stromal tumors (GISTs).METHODS: A total of 55 cases of GISTs were studied for the expression of c-Kit by immunohi...AIM: To investigate the prognostic significance of c-Kitgen emutation and DNA ploidy in gastointestinal stromal tumors (GISTs).METHODS: A total of 55 cases of GISTs were studied for the expression of c-Kit by immunohistochemistry, and the c-Kit gene mutations in exons 9, 11, 13, and 17 were detected by polymerase chain reaction-single strand confirmation polymarphism (PCR-SSCP) and denaturing high performance liquid chromatography (D-HPLC) techniques. DNA ploidy was determined by flow cytometry.RESULTS: Of the 55 cases of GISTs, 53 cases (96.4%) expressed c-Kit protein. The c-Kit gene mutations of exons 11 and 9 were found in 30 (54.5%) and 7 cases (12.7%),respectively. No mutations were found in exons 13 and 17.DNA aneuploidy was seen in 10 cases (18.2%). The c-Kit mutation positive GISTs were larger in size than the negative GISTs. The aneuploidy tumors were statistically associated with large size, high mitotic counts, high risk groups, high cellularity and severe nuclear atypia, and epithelioid type.There was a tendency that c-Kit mutations were more frequently found in aneuploidy GISTs.CONCLUSION: DNA aneuploidy and c-Kit mutations can be considered as prognostic factors in GISTs.展开更多
基金the Natural Science Foundation of Shaanxi Province, China （2004C115）.
文摘The aim of this study was to elucidate the expression and regulation of the c.kit protein in spermatogenesis of locusts. Immunohistochemistry and biological statistics were used to investigate the expression of the c-kit protein in four representative phases of spermatogenesis of three dominant species of locusts of Arcypteridae （Orthoptera： Acridoidea）, namely, Omocestus viridulus （Linnaeus）, Euchorthippus unicolor （Ikonn.）, and Euchorthippus vittatus Zheng, and so on, in Siping area of Jilin Province, China. The results revealed the following： （1） There was weak positive expression of the c-kit protein in spermatogonia and the positive granules were thinner; （2） there was a strong positive expression of the c-kit protein in primary spermatocyte and the positive granules became the largest than in all developmental stages; （3） the c-kit protein positive expression became stronger in secondary spermatocyte, while the positive granules became thinner; （4） there was strong positive expression of the c-kit protein and the positive granules were thinner in mature sperm, which were distributed on its head and tail; （5） there were strong positive protein granules massing at the end of spermary; （6） the positive intensity of the c-kit protein in spermatogenesis was significantly different among different species of locusts. The data suggested that the c-kit protein may play a crucial role in spermatogenesis as well as maintain the physiological action of sperms and fertilization, regulate the developmental speed of spermatogenesis, and/or maintain species isolation, etc.
文摘The tyrosine kinase receptor III, c-Kit/stem cell factor receptor and its ligand, human stem cell factor (huSCF) are the predominant regulator of mitogenesis in the hematopoietic stem and progenitor cells. However, gain-of-function mutations alter c-Kit auto-regulatory mechanisms to aberrant c-Kit signaling, leading to the onset or progression of cancerous transformations. The most common mutation of c-Kit is the substitution of aspartic acid residue in position 816 to valine (D816V), which is majorly responsible for its ligand-independent constitutive activation, and is implicated in hematopoietic malignancies. Currently, molecular targeted therapy is increasingly becoming a hot spot due to its specificity and low toxicity. As the molecular mechanisms responsible for D816V-c-Kit mediated tumorogenicity are largely unknown, in this study, we aimed to investigate the D816V-c-Kit signaling mediated downstream molecular targets. Specifically, we created c-Kit active mutant form D816V and performed inducible gene expression of mutant D816V-c-Kit in monomyelocytic cell line U937. Mutant D816V-c-Kit expressing cells revealed significantly enhanced cellular mitogenic activity compared to wild-type c-Kit expressing cells independent of huSCF. To examine the molecular targets regulating tumorogenic proliferation, we evaluated the consequences of mutant D816V-c-Kit expression on downstream gene expression profile by high throughput microarray technology. The levels of some of the relevant genes (PIK3CB, eIF4B, PRKCDBP, MOAP1) were validated by quantitative polymerase chain reaction. SLA, STAT5B, MAP3K2 and MAPK14 emerged as important downstream molecular targets of mutant D816V-c-Kit. Further, by dissecting the signaling pathways, we also demonstrated that the D816V-c-Kit mediated hematopoietic cell proliferation is dependent on molecular target p38 MAP kinase.
文摘AIM: To investigate the prognostic significance of c-Kitgen emutation and DNA ploidy in gastointestinal stromal tumors (GISTs).METHODS: A total of 55 cases of GISTs were studied for the expression of c-Kit by immunohistochemistry, and the c-Kit gene mutations in exons 9, 11, 13, and 17 were detected by polymerase chain reaction-single strand confirmation polymarphism (PCR-SSCP) and denaturing high performance liquid chromatography (D-HPLC) techniques. DNA ploidy was determined by flow cytometry.RESULTS: Of the 55 cases of GISTs, 53 cases (96.4%) expressed c-Kit protein. The c-Kit gene mutations of exons 11 and 9 were found in 30 (54.5%) and 7 cases (12.7%),respectively. No mutations were found in exons 13 and 17.DNA aneuploidy was seen in 10 cases (18.2%). The c-Kit mutation positive GISTs were larger in size than the negative GISTs. The aneuploidy tumors were statistically associated with large size, high mitotic counts, high risk groups, high cellularity and severe nuclear atypia, and epithelioid type.There was a tendency that c-Kit mutations were more frequently found in aneuploidy GISTs.CONCLUSION: DNA aneuploidy and c-Kit mutations can be considered as prognostic factors in GISTs.
