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目的:探讨上调子宫内膜癌细胞系HEC-1A中胰岛素样生长因子结合蛋白相关蛋白1的表达对子宫内膜癌细胞系HEC-1A增殖,凋亡及细胞周期的影响。方法:采用脂质体介导重组质粒转染HEC-1A细胞,按不同处理分为实验组(转染pEX-2-IGFBP7)、阴性对照组(转染pEX-2-Empty)、空白对照组(只加转染试剂),应用qRT-PCR技术检测各组细胞中IGFBP-rP1mRNA的表达水平,利用蛋白印迹法(Western Blot)测定各组IGFBP-rP1蛋白表达。采用四甲基偶氮唑蓝(MTT)法检测各组细胞系增殖抑制率,采用流式细胞仪检测各组凋亡率及细胞周期变化。结果:实验组、阴性对照组和空白对照组HEC-1A细胞中IGFBP-rP1mRNA的相对拷贝量分别为2.699±0.293,1.296±0.169,1.031±0.301,实验组明显高于阴性对照组(P=0.001)及空白对照组(P=0.000)。实验组IGFBP-rP1蛋白相对表达量(1.126±0.074)与阴性对照组(0.889±0.040,P=0.006)及空白对照组(0.884±0.043,P=0.005)中相比,差异有统计学意义。实验组24、48和72 h增殖抑制率(0.373±0.054,0.399±0.047,0.380±0.053)与阴性对照组(0.036±0.006,0.040±0.005,0.0334±0.006)和空白对照组(0.027±0.003,0.032±0.002,0.024±0.002)相比,差异均有统计学意义(P<0.01)。而阴性对照组与空白对照组增殖抑制率相比,差异无统计学意义(P>0.05)。转染后24 h,实验组凋亡率为(20.667±2.055)%,与阴性对照组[(3.967±0.351)%]和空白对照组[(4.967±0.252)%]相比,差异有统计学意义(P<0.01);实验组S+G2/M期细胞的比例为(30.980±1.461)%,明显低于阴性对照组[(38.797%±2.419)%]和空白对照组[(37.800%±0.350)%],差异有统计学意义(P<0.01)。结论:上调人子宫内膜癌细胞HEC-1A中IGFBP-rP1的表达可以抑制该细胞系的增殖,促进其凋亡,阻滞细胞周期。
AIM: To investigate the effects of up-regulating the expression of insulin-like growth factor-binding protein-1 (IGFBP-1) in human endometrial carcinoma cell line HEC-1A on the proliferation, apoptosis and cell cycle of endometrial carcinoma cell line HEC-1A. Methods: HEC-1A cells were transfected with liposome-mediated recombinant plasmids and divided into experimental group (transfected with pEX-2-IGFBP7), negative control group (transfected with pEX-2-Empty) and blank control group The expression of IGFBP-rP1 mRNA in each group was detected by qRT-PCR. The protein expression of IGFBP-rP1 in each group was detected by Western Blot. The proliferation inhibition rate of each group was detected by MTT assay. The apoptosis rate and cell cycle of each group were detected by flow cytometry. Results: The relative copy numbers of IGFBP-rP1 mRNA in experimental group, negative control group and blank control group were 2.699 ± 0.293, 1.296 ± 0.169 and 1.031 ± 0.301 respectively, which were significantly higher in experimental group than those in negative control group (P = 0.001 ) And blank control group (P = 0.000). The relative expression level of IGFBP-rP1 protein in the experimental group (1.126 ± 0.074) was significantly lower than that in the negative control group (0.889 ± 0.040, P = 0.006) and the blank control group (0.884 ± 0.043, P = 0.005) The inhibitory rates of proliferation in experimental group at 24, 48 and 72 h were significantly higher than those in control group (0.036 ± 0.006,0.040 ± 0.005,0.0334 ± 0.006) and blank control group (0.027 ± 0.003, 0.032 ± 0.002,0.024 ± 0.002), the differences were statistically significant (P <0.01). However, there was no significant difference between the negative control group and the blank control group (P> 0.05). At 24 h after transfection, the apoptosis rate in the experimental group was (20.667 ± 2.055)%, which was statistically significant compared with the negative control group [(3.967 ± 0.351)%] and the blank control group (4.967 ± 0.252%) (P <0.01). The proportion of cells in S + G2 / M phase in experimental group was (30.980 ± 1.461)%, which was significantly lower than that in negative control group (38.797% ± 2.419%) and blank control group (37.800% ± 0.350)%], the difference was statistically significant (P <0.01). Conclusion: Up-regulation of IGFBP-rP1 expression in human endometrial carcinoma cell line HEC-1A can inhibit the proliferation, promote the apoptosis and block the cell cycle.