Wip-1基因在子宫内膜癌组织中的表达及其与细胞凋亡的关系研究

Expression of Wip-1 gene in endometrial carcinoma and its correlation with apoptosis

目的探讨野生型p53诱导磷酸酶1(Wip-1)基因在子宫内膜癌组织中的表达及与细胞凋亡的关系。方法选取子宫内膜癌患者68例,子宫内膜不典型增生患者30例,并留取正常子宫内膜组织标本40份。培养子宫内膜癌细胞系ECC-1(ER阳性)和KLE(ER阴性),将细胞分为Wip-1siRNA组、siRNA对照组和空白对照组。利用实时荧光定量聚合酶链式反应(PCR)技术检测不同组织及不同转染组细胞中Wip-1基因表达,四甲基噻唑蓝(MTT)比色法检测不同处理组细胞增殖能力,Annexin V-异硫氰酸荧光素(FITC)/碘化丙啶(PI)双染色检测不同处理组细胞凋亡及细胞周期。结果子宫内膜癌组织中Wip-1mRNA表达水平(2.37±0.16)高于子宫内膜不典型增生组织(1.59±0.14)和正常子宫内膜组织(1.14±0.12),且子宫内膜不典型增生组织高于正常子宫内膜组织,差异均有统计学意义(P<0.01);子宫内膜癌组织中Wip-1mRNA表达水平与病理分期、病理分级、淋巴结转移、p53有关(P<0.01);Wip-1siRNA组ECC-1细胞和KLE细胞在转染后24、48、96h细胞存活率均低于siRNA对照组和空白对照组,差异均有统计学意义(P<0.05);Wip-1siRNA组ECC-1细胞和KLE细胞G0+G1期比例均高于siRNA对照组和空白对照组,而S期和G2+M期比例均低于siRNA对照组和空白对照组,差异均有统计学意义(P<0.01);Wip-1siRNA组ECC-1细胞和KLE细胞凋亡率均高于siRNA对照组和空白对照组,差异均有统计学意义(P<0.05)。结论 Wip-1基因参与了子宫内膜癌细胞增殖和凋亡过程。

Objective To investigate the expression of Wip-1 gene in endometrial carcinoma and its correlation with apoptosis. Methods Totally 68 cases of patients with endometrial carcinoma,30 cases of patients with endometrial atypical hyperplasia and 40 cases of normal endometrium specimens were selected. The endometrial carcinoma cell＇ line ECC-1 （ER-positive） and KLE （ER-negative） were cultured. The cells were divided into Wip-1 siRNA group, siRNA-control group and blank control group. The expressions of Wip-1 in different tissues and different transfected cells were detected by real-time fluorescence quantitative PCR. The cell proliferations in different groups were tested by MTT assay. The cell apoptosis and cell cycle of the different transfected groups were detected by Annexin V-FITC/PI double staining method. Results The expression level of Wip-1 mRNA in endometrial carcinoma tissues were（2.37±0. 16）, which were higher than that of atypical endometrial hyperplasia tissues and normal endometrial tissues,which were（1.59±0.14） and（1. 14± 0. 12）, respectively, and that of atypical endometrial hyperplasia tissues were higher than that of normal endometrium tissues, the differences were statistically significant （P〈0.01）. The expression level of Wip-1 mRNA in endometrial carcinoma tissues were related with pathological stage, pathological grade,lymph node metastasis and p53 （P%0. 001）. The cell survival rates after transfected 24,48,96 h of ECC-1 cells and KLE cells in Wip-1 siRNA group were lower than those in siRNA-control group and blank control group,the differences were statistically significant（P%0.05）. The proportions of G0 ＋ G1 phase of ECC-1 cells and KLE cells in Wip-1 siRNA group were higher than those in siRNA-control group and blank control group,while the proportions of S stage and G2＋M stage were lower than those in siRNA-control group and blank control group, the differences were statistically significant（P〈0.01）. The cell apoptosis rates of ECC-1 cells and KLE cells in Wip- 1 siRNA group were higher than those in siRNA-control group and blank control group, the differences were statistically significant （P〈0.05）. Conclusion Wip-1 gene is involved in the processes of proliferation and apoptosis in endometrial carcinoma cells.