摘要
目的探讨Ras/Raf/MAPK信号通路及其通路下游靶基因CyclinD1与病理性瘢痕癌变的相关性。方法 (1)激光扫描共聚焦显微技术,对病理性瘢痕和瘢痕癌组织进行K-ras、H-ras、N-ras免疫荧光双标记。(2)免疫组织化学SP法分别检测正常皮肤、病理性瘢痕和瘢痕癌三组组织中MAPK、Cy-clinD1蛋白的表达。(3)原位杂交技术检测三组组织中MAPK mRNA、CyclinD1 mRNA的表达。(4)用基因测序技术,检测病理性瘢痕上皮中K-ras、H-ras、N-ras第12、13位密码子突变。结果 (1)免疫荧光双标记:K-ras、H-ras、N-ras在病理性瘢痕上皮中呈现较弱荧光,为弱阳性表达;在瘢痕癌组织中呈现较强荧光,为强阳性表达。(2)MAPK及其mRNA和CyclinD1及其mRNA在正常皮肤表皮均呈阴性或弱阳性表达,在皮肤病理性瘢痕上皮中呈弱阳性表达,在瘢痕癌组织中呈强阳性表达。瘢痕癌组表达水平(阳性面积)、表达强度(平均吸光度)与正常皮肤、病理性瘢痕组比较,差异均有统计学意义(P<0.01);但正常皮肤组与病理性瘢痕组比较,差异无统计学意义(P>0.05)。(3)在病理性瘢痕中未发现K-ras、H-ras、N-ras第12、13位密码子突变。结论 (1)Ras、MAPK、CyclinD1蛋白及其MAPK mR-NA、CyclinD1 mRNA不是瘢痕上皮癌变的早期信号。(2)K-ras、H-ras、N-ras第12、13位密码子突变与病理性瘢痕上皮癌变无关。
Objective To detect the function of Ras/Raf/MAPK pathway and the downstream target gene CyclinD1 in the carcinogenesis of cutaneous pathologic scar. Methods （1） We adopted immunofluores cence dual staining for K-ras, H-ras and N-ras in the pathologic scar and carcinoma of scar, respectively, and observed under laser confocal microscopy; （2） Immunohistochemistry （IHC） for MAPK and Cy clinD1 were used for normal skin, pathologic scar and carcinoma of scar, respectively; （3） In situ hybrid ization （ISH） was used for mRNA detection of MAPK and CyclinD1 ; （4） The 12th and 13th codon muta tions of K-ras, H-ras and N-ras were detected by genetic sequencing. Results （1） The dual labeling of immunofluorescence of K-ras, H-ras and N-ras showed weak positive in pathologic scar, while the carci- noma of scar were strong positive. （2） ISH for mRNA of MAPK and CyclinD1 showed negative or weak positive in the normal epidermis and pathological scar, but strong positive in the carcinoma of scar. The expression level（positive area） and intensity （average optical density） between the carcinoma group and normal skin group or pathologic scar group were statistical different （P〈0. 01）; but without statistical difference between normal skin group and pathologic scar group （P〉0. 05）. （3） Genetic sequencing did not show mutation on the 12th or 13th codon. Conclusion （1） Ras, MAPK and CyclinD1 were not early signals for carcinogenesis of patho｜ogic scar. （2） The 12th or 13th codon mutation of K-ras, H-ras and N- ras was not related with the carcinogenesis of pathologic scar.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2013年第7期675-681,共7页
Cancer Research on Prevention and Treatment
基金
合肥医学院自然科学类招标课题资助项目(F-552)
贵州省科技攻关项目资助课题(2010-3080)
