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目的: 探讨在肿瘤细胞中凋亡与增殖平衡的变化(apoptotic index/proliferative index,AI/PI)以及与相关基因的关系. 方法:取普外科手术切除的标本68例,其中胃正常组10例,癌旁组10例,胃癌标本48例.术前均未接受过化疗或放疗.男32例,女16例.年龄24-74(平均54.4)岁. 术前内镜活检病检诊断,术后病理诊断证实.标本均经中性甲醛(40 g/L)固定,常规石蜡包埋,作4μm厚连续切片.应用流式细胞仪,分别定量分析细胞的凋亡群体和增殖群体;免疫组化方法检测survivin,Fas与FasL基因表达的变化;共聚焦显微镜对细胞增殖进行形态学观察. 结果:在从胃正常组,癌旁组到肿瘤组细胞的演进过程中,AI/PI分别为0.46±0.14,0.35±0.12,0.23±0.11. 胃正常组,癌旁组和肿瘤组细胞相互之间,差异有显著性(F=19.453,P=0.000<0.01).胃癌细胞的AI/PI经频数分布表分析和检验均服从正态分布.胃癌细胞AI/PI 范围0.05-0.46.中位数和均数±标准差分别为0.245和0.23±0.11.Skewness的值为0.067,Kurtosis的值为-0.868. 胃癌细胞随着恶性进展,AI/PI总体呈下降的趋势, 与淋巴转移和远处转移相关(P<0.05).survivin,Fas与Fasl,基因表达阳性染色主要定位于肿瘤细胞胞质或胞膜中,为粗细不一的棕黄色颗粒.在48例胃癌中survivin,Fas与FasL的阳性表达率分别为56.2%,43.8% 和60.4%.AI/PI的中位数为0.245,将胃癌分为>0.245 组和<0.245组.结果显示相应的两组组织切片中survivin 阳性表达率增加(X2=6.857,P<0.01),而Fas的阳性表达率降低(x2=4.148,P<0.05)和FasL的阳性表达率增加(x2=4.269,P<0.05),.共聚焦显微镜显示肿瘤组织中Ki- 67高表达. 结论:AI/PI总体呈下降的趋势,可能与survivin基因表达上调,Fas基因表达下调有关.
Objective: To explore the relationship between apoptosis index and proliferative index (AI / PI) and related genes in tumor cells.Methods: Sixty-eight surgical specimens of general surgery were obtained, of which 10 , Paraneoplastic group of 10 cases, gastric cancer specimens of 48. Preoperative chemotherapy or radiotherapy have not received 32 cases of male and 16 females, aged 24-74 (average 54.4 years old.) Preoperative biopsy biopsy diagnosis, surgery Pathological diagnosis was confirmed.The specimens were fixed with neutral formaldehyde (40 g / L), embedded in paraffin and cut into 4μm thick sections.Apoptotic and proliferative cells were quantified by flow cytometry The changes of the expression of survivin, Fas and FasL genes were detected by confocal microscopy, and the morphological changes were observed by confocal microscopy.Results: The AI / PI of the cells from the normal gastric cancer group to the adjacent cancer group were 0.46 ± 0.14,0.35 ± 0.12,0.23 ± 0.11.The difference of AI / PI in gastric cancer cells, adjacent cancer group and tumor group was significant (F = 19.453, P = 0.000 <0.01) Distribution table analysis and test obey the normal distribution of gastric cancer AI / PI range of 0.05-0.46 median and mean ± standard deviation were 0.245 and 0.23 ± 0.11 respectively, the value of Skewness was 0.067, and the value of Kurtosis was -0.868. As the malignant progression of gastric cancer cells, the AI / PI tended to decrease, which was correlated with lymphatic metastasis and distant metastasis <0.05) .Survivin, Fas and Fasl gene expression positive staining mainly located in the cytoplasm of tumor cells or cell membrane, as the thickness of the brown granules.In 48 cases of gastric cancer, the positive rates of survivin, Fas and FasL were 56.2%, 43.8% and 60.4%, respectively.The median of AI / PI was 0.245.The gastric cancer was divided into> 0.245 group and <0.245 group.The results showed that the positive expression rate of survivin in the corresponding two groups of tissues increased (X2 = 6.857 , P <0.01), while the positive rate of Fas decreased (x2 = 4.148, P <0.05) and FasL increased (x2 = 4.269, P <0.05) .Confocal microscopy showed that the expression of Ki- 67 High expression.Conclusion: AI / PI generally showed a downward trend, which may be related to up-regulation of survivin gene and down-regulation of Fas gene expression.