摘要
目的:探讨PI3K/AKT信号传导通路中PI3K、AKT及其下游靶基因编码的Bcl-2蛋白与瘢痕癌癌细胞凋亡的相关性。方法:以15例病理性皮肤瘢痕被覆上皮和20例皮肤瘢痕癌组织为研究对象,以10例正常皮肤表皮组织为对照。这些标本均为石蜡包埋组织,源于遵义医学院附属第一和第五医院病理科2006-01-01-2011-12-30的外检标本。1)采用免疫组织化学技术(SP法)分别检测以上3组组织中PI3K、AKT和Bcl-2蛋白的表达。2)采用核酸分子原位杂交技术检测PI3K mRNA、AKT mRNA的表达。3)以原位末端标记法(Tunel法)检测细胞的凋亡水平。4)免疫组化和原位杂交图像运用图像分析软件计算其平均光密度和阳性面积,Tunel图像计算其凋亡指数,所得数据运用SPSS 16.0软件包进行统计学分析。结果:1)PI3K蛋白和PI3K mRNA在正常皮肤表皮、病理性瘢痕上皮和瘢痕癌组织中分别呈阴性、弱阳性和阳性表达。瘢痕癌组PI3K表达水平为0.172±0.039,表达强度为0.301±0.045,与正常皮肤组0.004±0.003和0.185±0.021,病理性瘢痕组0.011±0.009和0.203±0.034比较,差异有统计学意义,P<0.01;瘢痕癌组PI3KmRNA表达水平为0.137±0.037,表达强度为0.379±0.054,与正常皮肤组0.008±0.007和0.265±0.016,病理性瘢痕组0.027±0.012和0.293±0.031比较,差异有统计学意义,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。2)AKT蛋白在瘢痕癌组表达水平为0.168±0.052,表达强度为0.388±0.081,与正常皮肤组0.005±0.005和0.218±0.036,病理性瘢痕组0.028±0.025和0.282±0.049比较,差异有统计学意义,P<0.01;AKT mRNA在瘢痕癌组表达水平为0.144±0.032,表达强度为0.345±0.031,与正常皮肤组0.017±0.004和0.244±0.025,病理性瘢痕组0.037±0.019和0.257±0.027比较,差异有统计学意义,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。3)Bcl-2在3组组织中均呈阴性表达。其表达水平、表达强度在正常皮肤组(0.006±0.003和0.168±0.019)、病理性瘢痕组(0.008±0.005和0.175±0.027)和瘢痕癌组(0和0.167±0.015)之间比较,差异无统计学意义,P=0.136。4)正常皮肤组和病理性瘢痕组AI分别为(8.48±0.78)%和(8.14±0.53)%,高于瘢痕癌组(6.41±0.60)%,且差异有统计学意义,P=0.01;但正常皮肤组与病理性皮肤瘢痕组比较,差异无统计学意义,P=0.129。结论:PI3K/AKT信号传导通路的抗凋亡途径,可能是通过Bcl-2以外的其他效应分子来实现的。
OBJECTIVE： To explore whether the PI3K and AKT in PI3K/AKT signaling pathway and the pathway downstream proteins Bcl-2 are correlated with the cell apoptosis in scar carcinoma. METHODS： Using 10 normal skin epi- dermis as control, we observed 15 pathological skin scar epithelium and 20 skin scar carcinoma tissues. All the paraffin- embedded tissue were collected from the First and Fifth Affiliated Hospitals of Zunyi Medical College during the period 2006-01-01--2011-12-30. The expression of PI3K, AKT, Bel-2 were detected by SP immunohistochemieal staining tech- nique. The mRNA expression levels of PI3K and AKT were detected by in situ hybridization （ISH）. Apoptotic bodies were detected by terminal deoxynucleo- tidyltransferase （TdT）-mediated dUTP nick end labeling （Tunel assay）. We cal- culated the average optical density and positive area of IHC and ISH images and the apoptotic index of Tunel images using image analysis software. All statistical analyses were done with SPSS16.0 software. RESULTS： PI3K protein and its mR- NA in normal skin epidermis, pathological skin scar epithelium and sear carcinoma tissues differently showed negative ex- pression, weakly positive expression and positive expression. The expression level and intensity of sear carcinoma group （0. 172±0. 039 and 0. 137±0. 037;0. 301＋0. 045 and 0. 3794-0. 054） ,were significantly different from the pathological skin scar group（0. 011±0. 009 and 0. 027±0. 012;0. 2034±0. 034 and 0. 293＋-0. 031） and the normal skin group（0. 004± 0. 003 and 0. 008±0. 007; 0. 185±0. 021 and 0. 265±0. 016, P〈0.01）. There was no significant difference of expression level between the normal skin group and the pathological skin scar group （P〈0.05）. AKT protein and AKT mRNA in normal skin epidermis, pathological skin scar epithelium and scar carcinoma tissues differently showed negative expres- sion,weakly positive expression and strongly positive expression. The expression level and intensity of scar carcinoma group （0. 1684±0. 052 and 0. 1444±0. 032; 0. 3884-0. 081 and 0. 345±0. 031） were significantly different from the normal skin group（0.0054±0.005 and 0.017±0.004;0. 218±0. 036 and 0. 244~0.025） and the pathological skin scar group （0.0284±0.025 and 0.0374±0.019;0.282＋-0.049 and0.257±0.027, P〉0.01）. There was no significant difference be- tween the expression of the normal skin group and the pathological scar group （P〈0.05）. Bel-2 showed negative expres- sion in three groups of organizations. The expression of level and intensity in normal skin group （0. 006 4± 0. 003 and 0. 1684±0. 019）, pathological skin scar group（0. 0084-0. 005 and 0. 175±0. 027）and scar carcinoma group （0 and 0. 167± 0. 015） were not significant （P=0. 136）. Apoptotic index of the normal skin （8. 484±0.78）% and the pathological skin scar group（8.14 4± 0.53）% was significantly higher than that of the scar carcinoma group （6.41 4± 0.60）% （P = 0.01 ）, but there was no significant difference of apoptotic index between the normal skin group and the pathological skin scar group （P=0. 129）. CONCLUSION： There may be other molecule instead of Bcl-2 working through the PI3K/AKT signa- ling pathway to achieve apoptosis inhibition.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2014年第8期599-604,共6页
Chinese Journal of Cancer Prevention and Treatment
基金
贵州省科技攻关项目(2010-3080)
