非小细胞肺癌细胞周期蛋白D_1、p16及视网膜母细胞瘤基因蛋白表达与预后的关系(英文)

Relationship between expressions of cyclinD_1, p16 and retinoblastoma gene protein and prognosis of non-small-cell lung cancer

背景:细胞周期调控失常是造成细胞异常增殖进而导致肿瘤发生的重要机制,p16、视网膜母细胞瘤基因蛋白参与的细胞周期蛋白D1/细胞周期蛋白依赖性激酶通路是一条重要的细胞周期调节途径。目的:了解在非小细胞肺癌中细胞周期蛋白D1、p16、视网膜母细胞瘤基因蛋白表达的情况,并探讨它们与预后之间的关系。设计:应用免疫组织化学方法检测相关指标的表达情况,利用随访资料进行生存率及预后因素的比较分析。单位:福建省立医院呼吸科。对象:实验于2002-01/06月在福建省立医院病理科完成。肺癌组织标本来自68例非小细胞肺癌患者,肺癌组织学分型依据1981年WHO制定的标准,TNM分期依据1997年国际抗癌联盟修订的标准。方法:将肺癌组织标本应用免疫组织化学方法染色,用以知阳性染色的肺癌切片为阳性对照,用磷酸盐缓冲液替代一抗为阴性对照。以肿瘤细胞核或细胞质呈现棕黄色颗粒为阳性。每例标本随机观察5个高倍视野,每个视野计数200个肿瘤细胞,取平均值。根据阳性细胞所占比例分为:阳性细胞数小于5%或看不到明显的阳性细胞为“-”;阳性细胞数在5%～20%为“+”;阳性细胞数在21%～50%为“”;阳性细胞数超过50%为“”。检测68例非小细胞肺癌组织中细胞周期蛋白D1、p16、视网膜母细胞瘤基因蛋白表达情况。主要观察指标:细胞周期蛋白D1、p16、视网膜母细胞瘤基因蛋白表达情况及患者术后生存期。结果:①非小细胞癌组住中细胞周期蛋白D1、p16、视网膜母细胞瘤基因蛋白表之间的相关性:达率分别为55.9%,48.5%,52.9%,应用χ2对资料进行分析,其表达之间均无明显相关。②细胞周期蛋白D1、p16、视网膜母细胞瘤基因蛋白表达与非小细胞肺癌患者预后的关系:细胞周期蛋白D1阳性表达者的平均生存时间较细胞周期蛋白p16阴性者短[(12.40,35.40)个月,(χ2=5.31,P=0.021)];p16阳性表达者的平均生存时间较p16阴性者长[(27.84,15.95)个月,(χ2=4.38,P=0.036)];视网膜母细胞瘤基因蛋白阳性达者的平均生存时间与视网膜母细胞瘤基因蛋白阴性者相比无显著差别[(18.50,24.19)个月,(χ2=0.04,P>0.05)]。③非小细胞肺癌患者预后多因素Cox风险比例模型分析:提示细胞周期蛋白D1过度表达者预后差(P=0.057),而p16则为1个预后保护性因素(P=0.039)。结论:与细胞周期G1→S调控相关的基因细胞周期蛋白D1、p16、视网膜母细胞瘤基因均参与了非小细胞肺癌的发生发展,检测细胞周期蛋白D1、p16的表达水平对预测预后有重要意义。

BACKGROUND： Disturbed regulation of cell cycle is the essential mechanism on abnormal proliferation of cell and incidence of cancer. CyclinD1/ cyclin-dependent-kinase pathway involved with p16 and retinoblastoma gene protein is an important approach on regulation of cell cycle. OBJECTIVE： To understand the expressions of cyclinD1, p16 and retinoblastoma gene protein in non-small-cell lung cancer and probe into the relationship with the prognosis of cancer. DESIGN： Immunohistochemical assay was used to measure the expres- sions of relevant indexes and follow-up data were utilized for comparative analysis on survival rate and prognostic factor. SETTING： Department of Respiratory Disease, Fujian Provincial Hospital PARTICIPANTS： The experiment was performed in Department of Pathology of Fujian Provincial Hospital from January to June 2002. Tissue specimens of lung cancer were from 68 cases of non-small-cell lung cancer. Histological classification of lung cancer was based on the standards implemented by WHO in 1981 and TNM staging was on the standards revised by International Union Against Cancer （UICC） in 1997. METHODS： Tissue specimens of lung cancer were stained with immunohistochemical method, the positive-stained splices of lung cancer were taken as positive control and those with 1^st antibody replaced by phosphate buffer were taken as negative control. The positive was identified if tumor cell nucleus or cytoplasm presented brown-yellow granules. 5 high-power visual fields were observed at randomize in each specimen and 200 tumor cells were counted and the average was obtained. According to the pereeptions of positive cell, in which, positive cell count was less by 5% or no obvious positive cell appeared “-”, positive cell count was in range from 5% to 20% “＋”, positive cell count was in range from 21% to 50% “＋＋” and positive cell count was over 50% “＋＋＋” the expression of cyclin D1, p16 and retinoblastoma gene protein in 68 cases of non-small-cell lung cancer were assayed. MAIN OUTCOME MEASURES： Expressions of cyclinD1, p16 and retinoblastoma gene protein and survival phase of patients after operation. RESULTS： ① Correlation among the expressions of cyclinD1, p16 and retinoblastoma gene protein： The expressive rates were 55.9%, 48.5% and 52.9% respectively. χ^2 was applied for analysis on the data and it was indicated that there was no remarkable correlation among their expressions. ② Relationship between expressions of cyclinD1, p16 and retinoblastoma gene protein and prognosis of non-small-cell lung cancer： The average survival phase of cyclinD1 positive expression was shorter than that of negative expression [（12.40, 35.40） months, （χ^2=5.31 ,P =0.021）]. The average survival phase of positive case of p16 positive expression was longer than that of negative one [（27.84, 15.95） months, （χ^2=4.38,P =0.036）]. The average survival phase of positive expression of retinoblastoma gene protein was not more different significantly from the negative one [（18.50, 24.19） months, （χ^2=0.04,P 〉 0.05）]. ③ Multi-factor analysis on prognosis of non-small-cell lung cancer with Cox proportional hazard model： It was indicated that the prognosis of excessive expression of cyclinD1 was poor （P=0.057） and that of p16 only presented 1 prognostic protective factor （P=0.039）. CONCLUSION： CyclinD1, p16 and retinoblastoma gene associated with cyclinG1-S regulation are all involved in incidence and development of non-small-cell lung cancer. The assay of expressions of cyclinD1 and p16 is essentially significant for prognosis.