miR-375靶向IGF-1对子宫内膜癌细胞增殖的调控作用

The effects of miR-375 targeting IGF-1 on the proliferation of endometrial carcinoma cells

目的探讨miR-375靶向胰岛素样生长因子-1(IGF-1)对子宫内膜癌细胞增殖的调控作用。方法选择对数生长期的人子宫内膜癌Ishikawa细胞,随机分为5组,模拟剂组转染miR-375模拟剂,模拟剂NC组转染miR-375模拟剂NC,抑制剂组转染miR-375抑制剂,抑制剂NC组miR-375抑制剂NC,对照组为未转染的Ishikawa细胞。利用qRT-PCR技术验证miR-375的表达;利用CCK8及凋亡实验检测Ishikawa细胞增殖及凋亡情况;利用Western blot技术检测IGF-1表达水平。结果转染后48 h与96 h,模拟剂组比模拟剂NC组细胞的增殖能力下降,抑制剂组比抑制剂NC组细胞的增殖能力增强(P<0.05)。转染48 h后,模拟剂组、miR-375 NC组、抑制剂组、抑制剂NC组、对照组凋亡率分别为(40.00±3.18)%、(18.19±2.01)%、(7.44±1.11)%、(18.11±1.86)%、(17.22±3.84)%,miR-375 NC组显著低于模拟剂组,抑制剂NC组显著高于抑制剂组(P<0.05)。转染48 h后,IGF-1蛋白在模拟剂组、miR-375 NC组、抑制剂组、抑制剂NC组、对照组中的相对表达水平分别为0.45±0.22,1.45±0.14,3.89±0.11,1.66±0.33,1.74±0.32,miR-375 NC组显著高于模拟剂组,抑制剂NC组显著低于抑制剂组(P<0.05)。结论过表达的miR-375能抑制子宫内膜癌细胞增殖与促进细胞凋亡,其具体机制可能是miR-375通过负向调控IGF-1的表达而实现。

Objective To investigate the effects of miR-375 targeting insulin like growth factor-1（IGF-1） on the proliferation of endometrial carcinoma cells. Methods The human endometrial cancer Ishikawa cells were randomly divided into 5 groups,the mimic group transfected miR-375 mimic,the mimic NC group transfected miR-375 mimic NC,the inhibitor group transfected miR-375 inhibitor,and the control group was untransfected Ishikawa cells; and were to detect of Ishikawa cell proliferation and apoptosis; the expression level of IGF-1 protein by detection of Ishikawa cells transfected with Western blotting technology. Results After transfection 48 h and 96 h,the proliferationin in the mimic group were lower than that of mimic NC group,and the proliferation ability of inhibitor group were higher than that of inhibitor NC group,and compared the difference were significant（P〈0. 05）. After transfection of 48 h,the apoptotic rates of mimic group,miR-375 NC group,inhibitor group,inhibitor NC group and control group were（40. 00 ± 3. 18） %,（18. 19 ± 2. 01） %,（7. 44 ± 1. 11） %,（18. 11 ± 1. 86） %,（17. 22 ± 3. 84） % respectively,and there was significant difference compared between them（P〈0. 05）. After transfection of 48 h,the relative expression level of IGF-1 protein in mimic group,miR-375 NC group,inhibitor group,inhibitor NC group and control group were 0. 45 ± 0. 22,1. 45 ± 0. 14,3. 89 ± 0. 11,1. 66 ± 0. 33,1. 74 ± 0. 32,respectively,and the difference was significantly（P〈0. 05）. Conclusion The over expression of miR-375 can promote cell apoptosis and inhibit cell proliferation. The specific mechanism may be that miR-375 suppresses the occurrence and development of endometrial cancer cells through negative regulation of IGF-1 expression.