目的:探讨胃肠间质瘤(gastrointestinal stomal tumors,GIST)患者肿瘤组织中c-Kit和PDGFRA基因各亚型突变情况的研究。方法:应用直接测序方法检测65例GIST石蜡组织中c-Kit基因9、11、1 3和1 7外显子和P D G F R A基因1 2和1 8外显子突...目的:探讨胃肠间质瘤(gastrointestinal stomal tumors,GIST)患者肿瘤组织中c-Kit和PDGFRA基因各亚型突变情况的研究。方法:应用直接测序方法检测65例GIST石蜡组织中c-Kit基因9、11、1 3和1 7外显子和P D G F R A基因1 2和1 8外显子突变情况。结果:G I ST肿瘤组织中c-K i t基因总突变率为63.08%(41/65),外显子9、11、13和17的突变率分别为23.08%(15/65)、35.38%(23/65)、1.54%(1/65)和3.08%(2/65);c-Kit基因各外显子之间双重突变共2例(3.08%),其中9外显子与11外显子双重突变1例(1.54%),11外显子与17外显子双重突变1例(1.54%);c-Kit基因11号外显子内双重突变2例(3.08%),三重突变1例(1.54%)。PDGFRA基因总突变率为3.08%(2/65),均为外显子18突变。c-Kit基因和PDGFRA基因双突变共存型1例(1.54%),为c-Kit基因13外显子V654L点突变和PDGFR A基因18外显子D842V点突变。结论:GIST患者中c-Kit基因存在较高的突变率,尤其为9和11外显子突变,其基因突变亚型分类能指导精准医学下伊马替尼的肿瘤靶向治疗,PD GF R A基因突变率和存在两种及其以上突变发生的发生概率虽低但不容忽视。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
Objective:To investigate the expression of C-Kit and PDGFRα and their correlation with chemotherapy resis-tance in ovarian serous carcinoma.Methods:We undertook SP immunohistochemical technique to examine the express...Objective:To investigate the expression of C-Kit and PDGFRα and their correlation with chemotherapy resis-tance in ovarian serous carcinoma.Methods:We undertook SP immunohistochemical technique to examine the expression of C-Kit and PDGFRα in 59 cases with ovarian serous carcinomas,using archival paraffin-embedded specimens.Then we observed the correlation with chemotherapy resistance.Results:C-Kit and PDGFRα immunostainings were observed posi-tively expressed in 57.63% and 66.10% cases.C-Kit expression was statistically correlated with the progression of disease after first-line chemotherapy(P < 0.05),but PDGFRα was not statistically significant(P > 0.05).There were great difference between of C-Kit and PDGFRα expressions in samples of different differentiated and clinical stages of ovarian serous carci-nomas(P < 0.01 or P < 0.05).Conclusion:C-Kit is statistically correlated with chemotherapy resistance,while PDGFRα is not correlated.展开更多
文摘目的:探讨胃肠间质瘤(gastrointestinal stomal tumors,GIST)患者肿瘤组织中c-Kit和PDGFRA基因各亚型突变情况的研究。方法:应用直接测序方法检测65例GIST石蜡组织中c-Kit基因9、11、1 3和1 7外显子和P D G F R A基因1 2和1 8外显子突变情况。结果:G I ST肿瘤组织中c-K i t基因总突变率为63.08%(41/65),外显子9、11、13和17的突变率分别为23.08%(15/65)、35.38%(23/65)、1.54%(1/65)和3.08%(2/65);c-Kit基因各外显子之间双重突变共2例(3.08%),其中9外显子与11外显子双重突变1例(1.54%),11外显子与17外显子双重突变1例(1.54%);c-Kit基因11号外显子内双重突变2例(3.08%),三重突变1例(1.54%)。PDGFRA基因总突变率为3.08%(2/65),均为外显子18突变。c-Kit基因和PDGFRA基因双突变共存型1例(1.54%),为c-Kit基因13外显子V654L点突变和PDGFR A基因18外显子D842V点突变。结论:GIST患者中c-Kit基因存在较高的突变率,尤其为9和11外显子突变,其基因突变亚型分类能指导精准医学下伊马替尼的肿瘤靶向治疗,PD GF R A基因突变率和存在两种及其以上突变发生的发生概率虽低但不容忽视。
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.
基金Supported by a grant from the Scientific Research Foundation of Health Department of Hubei Province (No.JX3C54)
文摘Objective:To investigate the expression of C-Kit and PDGFRα and their correlation with chemotherapy resis-tance in ovarian serous carcinoma.Methods:We undertook SP immunohistochemical technique to examine the expression of C-Kit and PDGFRα in 59 cases with ovarian serous carcinomas,using archival paraffin-embedded specimens.Then we observed the correlation with chemotherapy resistance.Results:C-Kit and PDGFRα immunostainings were observed posi-tively expressed in 57.63% and 66.10% cases.C-Kit expression was statistically correlated with the progression of disease after first-line chemotherapy(P < 0.05),but PDGFRα was not statistically significant(P > 0.05).There were great difference between of C-Kit and PDGFRα expressions in samples of different differentiated and clinical stages of ovarian serous carci-nomas(P < 0.01 or P < 0.05).Conclusion:C-Kit is statistically correlated with chemotherapy resistance,while PDGFRα is not correlated.
