胰岛素对子宫内膜癌细胞增殖和凋亡的影响

Effects of insulin on proliferation and apoptosis of endometrial carcinoma cell

目的探讨胰岛素对子宫内膜癌细胞系 Ishikawa3-H-12细胞增殖、凋亡和细胞周期的影响。方法应用免疫细胞化学方法和 RT-PCR 技术检测 Ishikawa3-H-12细胞胰岛素受体(INSR)蛋白和 mRNA 的表达。以不同浓度胰岛素作用 Ishikawa3-H-12细胞不同时间,采用四甲基偶氮唑蓝比色法、流式细胞仪检测细胞增殖、凋亡和细胞周期。结果 (1)Ishikawa3-H-12细胞 INSR 蛋白呈棕黄色阳性表达,并可见 INSR 基因的表达。(2)胰岛素以浓度和时间依赖的方式促进子宫内膜癌细胞增殖,1×10^(-4)mol/L 胰岛素作用48 h 时促增殖作用最显著,增殖率为(340.2±15.9)%,与对照细胞(以胰岛素浓度为0作为对照,为100%)比较,差异有统计学意义(P<0.05)。(3)胰岛素以浓度和时间依赖的方式使 Ishikawa3-H-12细胞中 G_0/G_1期细胞比例减少,S 期细胞比例增加,1×10^(-4) mol/L胰岛素作用72 h 时最显著,G_0/G_1期细胞比例为(27.7±2.5)%,S 期细胞为(55.2±1.4)%,分别与对照细胞[分别为(67.6±1.5)%、(15.7±1.0)%]比较,差异均有统计学意义(P<0.05);胰岛素对G_2/M 期细胞无影响(P>0.05)。(4)随着胰岛素浓度的增加,Ishikawa3-H-12细胞凋亡率逐渐下降。1×10^(-4)mol/L 胰岛素作用最显著,作用24、48、72、96 h 时的细胞凋亡率分别为(1.76±0.16)%、(1.70±0.15)%、(1.56±0.20)%、(1.31±0.24)%,分别与对照细胞[分别为(9.81±0.61)%、(9.93±1.44)%、(9.10±0.66)%、(10.30±1.20)%]比较,差异均有统计学意义(P<0.05)。结论胰岛素对子官内膜痛 Ishikawa3-H-12细胞具有促进增殖、抑制凋亡的作用。

Objective To study the expression of insulin receptor （INSR） in endometrial cell line Ishikawa3-H-12, and the effects of insulin on proliferation, cell cycle distribution and apoptosis of Ishikawa3-H-12 cells. Methods Immunocytochemistry and RT-PCR methods were used to investigate the expression of INSR in Ishikawa3-H-12 cells. The effects of insulin at different concentrations and different time on proliferation, apoptosis and cell cycle distribution of endometrial carcinoma cells were observed by methyl thiazolyl tetrazolium （MTT） assay and fluorescence-activated cell sorting technique. Results （ 1 ） We demonstrated the expression of INSR in Ishikawa3-H-12 cell line. （2） Incubation with insulin stimulated a dose- and time-dependent proliferation response in Ishikawa3-H-12 cells with the peak response occurring at 48 hours with 1 × 10 ^-4 mol/L insulin incubation [ proliferative rate： （340. 2 ± 15.9 ） %, vs control （ 100% ）, P 〈 0. 05 ] . （ 3 ） The percentage of Ishikawa3-H-12 cells at G0/G1 phase decreased and percentage at S phase increased significantly with insulin treatment in a dose- and time-dependent manner with the peak response occurring at 72 hours with 1 × 10^ -4 mol/L insulin incubation [ G0/G1 phase （ 27.7 ±2.5）%,S phase （55.2 ± 1.4）%, vs control： （67.6 ±1.5）% and （15.7 ± 1.0）%, P〈0.05], whereas that of G2/M phase did not show significant alteration. （4） The apoptosis rate of Ishikawa3-H-12 cells was decreased gradually with increasing concentration of insulin, while the alteration was time- independent. The peak response occurred with 1 × 10 ^-4 mol/L insulin incubation [ apoptosis rate ： （ 1.76 ± 0.16）%, （1.70±0.15）%, （1.56±0.20）%, （1.31±0.24）% when incubated at 24, 48, 72 and 96 hours respectively, vs control, P 〈 0. 05 ]. Conclusion Insulin can promote the proliferation and inhibit the apoptosis of endometrial carcinoma cells.