Ras、MAPK、Cyclin D1与皮肤瘢痕癌的相关性研究  

Correlation of Ras,MAPK and Cyclin D1 with skin scar cancer

作　　者：郭瑞珍[1] 王海青[1] 欧小波[1] 

机构地区：[1]遵义医学院珠海校区病理学教研室,珠海519041

出　　处：《临床与实验病理学杂志》2013年第4期393-397,共5页Chinese Journal of Clinical and Experimental Pathology

基　　金：贵州省科技攻关项目(2010-3080);遵义医学院自然科学类招标课题(F-552)

摘　　要：目的探讨Ras/Raf/MAPK信号通路和通路下游靶基因Cyclin D1与皮肤瘢痕癌的相关性。方法 (1)用激光扫描共聚焦显微技术对病理性瘢痕和瘢痕癌进行K-ras、H-ras、N-ras免疫荧光双标记;(2)提取DNA,检测病理性瘢痕和瘢痕癌组织中K-ras、H-ras、N-ras第12、13位密码子的突变;(3)采用免疫组化SP法检测正常皮肤、病理性瘢痕和瘢痕癌组织中MAPK、Cyc-lin D1蛋白的表达;(4)采用原位杂交技术检测3组组织中MAPK mRNA、Cyclin D1 mRNA的表达。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较弱荧光为弱阳性,在瘢痕癌组织中呈较强荧光为强阳性;(2)在病理性瘢痕和瘢痕癌中未发现K-ras、H-ras、N-ras第12、13位密码子突变;(3)MAPK和Cyclin D1的蛋白及mRNA在正常皮肤表皮均呈阴性或弱阳性,在皮肤病理性瘢痕上皮中呈弱阳性,在瘢痕癌组织中呈强阳性。瘢痕癌组表达水平(阳性面积)、表达强度(平均光密度)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),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)。结论 (1)Ras、MAPK、Cyclin D1基因的高表达与瘢痕癌的发生密切相关,各种基因共同发挥了协同作用;(2)K-ras、H-ras、N-ras第12、13位密码子突变与瘢痕癌的发生无相关性。Purpose To explore the correlation of Ras/Raf/MAPK signaling pathway and the pathway downstream target gene Cyclin D1 with the skin sear cancer. Methods （ 1 ） scanning confocal microscopy and immunofluorescence double labeling of the K-ras, H- ras and N-ras were conducted in the paraffin-embedded tissue sections of pathological scars and scar cancer. （2） DNA was extracted through skin pathological sear tissues and scar carcinoma tissues, and then H-ras, N-ras and K-ras mutations in codons 12 and 13 were analyzed using PCR and sequencing. （3） the expression of MAPK and Cyelin D1 protein was detected in normal skin, pathological scar and scar cancer tissues by the immunohistoehemieal method of SP. （4） the expression of MAPK mRNA and Cyclin D1 mRNA was detected in the three groups by in situ hybridization. Results （ 1 ） the laser scanning confocal microscopy showed that the expression of K-ras, H-ras and N-ras in pathological scar epithelium was weak; it showed strong expression in scar cancer. （2） the eodons 12 and 13 of H-ras, N-ras and K-ras genes did not have the mutation after sequencing in pathological scar and scar cancer. （3） MAPK and its mRNA and Cyelin D1 and its mRNA showed negative or weakly positive expression in normal skin epidermis, weakly positive expres- sion in pathological scar epithelium, but strongly positive expression in scar cancer. The expression of levels （positive area） and inten- sity （mean optical density） differences were statistically significant （P 〈 0.01 ） by compared with normal skin group and skin scar group. But the differences of expression was no statistically significant between normal skin group and pathological skin scar group（ P 〉 0. 05 ）. Conclusions The strong expression of Ras, MAPK and Cyelin D1 gene is closely related with the occurrence of scar carcino- ma, in which a variety of genes play a synergistic role. But the mutation of K-ras, H-ras and N-ras genes 12 and 13 codons has no cor- relation with the pathogenesis of s

关 键 词：瘢痕癌 信号传导通路 CYCLIN D1 免疫荧光 基因突变 免疫组织化学 

分 类 号：R739.5[医药卫生—肿瘤]