Wang X H, Zhang Y, Liu L Z, Shang C G
Department of Obstetrics and Gynecology,Peking University First Hospital, Beijing 100034, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2018 Oct 18;50(5):767-773.
To determine the effect of metformin and adiponectin on the proliferation of EC cells and the relationship between metformin and adiponectin.
The proliferation impact of different concentrations of metformin and adiponectin on two types of EC cells ishikawa (IK) and HEC-1B was confirmed by CCK-8 method. qRT-PCR and Western blot were used to detect the effect of different concentrations of metformin on the changes of adiponectin receptors (AdipoR1 and AdipoR2) of the EC cells both in mRNA and protein level and the role of compound C, an adenosine monophosphate-activated protein kinase (AMPK) inhibitor, on the above effects.
(1) Both metformin and adiponectin could significantly promote the proliferation of endometrial cancer (EC) cells in a time and concentration dependent manner (P<0.05).(2)Metformin and adiponectin had synergy anti-proliferative effect on EC cells and the combination index (CI) value of IK cells was 0.906 34 and of HEC-1B cells was 0.827 65. (3)qRT-PCR was used to detect the mRNA levels of AdipoR1 and AdipoR2 after 5 mmol/L and 10 mmol/L metformin, respectively, stimulating IK and HEC-1B cells for 48 hours and the mRNA expressions of AdipoR1 and AdipoR2 were significantly increased when compared with the control group (0 mmol/L)(IK: AdipoR1 of 5 mmol/L and 10 mmol/L group: P<0.001,AdipoR2 of 5 mmol/L group: P<0.001; HEC-1B: AdipoR1 of 5 mmol/L group: P<0.001, 10 mmol/L group: P=0.023, AdipoR2 of 5 mmol/L group: P<0.001, 10 mmol/L group: P=0.024). When combined with compound C, the RNA levels of AdipoR1 and AdipoR2 were not different compared with the control group (0 mmol/L, P>0.05). (4) Western blot was used to detect the protein levels of AdipoR1 and AdipoR2 after 5 mmol/L and 10 mmol/L metformin, stimulating IK and HEC-1B cells for 48 hours and the protein level was significantly increased when compared with the control group (0 mmol/L)(IK: AdipoR1 of 5 mmol/L group: P=0.04, 10 mmol/L group: P=0.033, AdipoR2 of 5 mmol/L group: P=0.044, 10 mmol/L group: P=0.046; HEC-1B: AdipoR1 of 5 mmol/L group: P=0.04, 10 mmol/L group: P=0.049, AdipoR2 of 5 mmol/L group: P=0.043, 10 mmol/L group: P=0.035). When combined with compound C,the protein levels of AdipoR1 and AdipoR2 were not different compared with the control group (0 mmol/L, P>0.05).
We find that metformin and adiponectin have synergy anti-proliferative effect on EC cells. Besides, metformin can increase adiponectin receptors expressions of EC cells both in mRNA and protein levels and this effect is accomplished by the activation of AMPK signaling pathway.
探讨二甲双胍和脂联素对子宫内膜癌细胞增殖的影响以及二者之间的关系。
采用CCK-8法检测不同浓度二甲双胍和脂联素对两种子宫内膜癌细胞系ishikawa（IK）和HEC-1B增殖的影响。运用qRT-PCR和蛋白质免疫印迹法检测不同浓度二甲双胍对子宫内膜癌细胞脂联素受体（AdipoR1和AdipoR2）mRNA和蛋白水平变化的影响，以及磷酸腺苷活化蛋白激酶（AMPK）抑制剂化合物C对上述作用的影响。
（1）二甲双胍和脂联素均能以时间和浓度依赖的方式显著促进子宫内膜癌（EC）细胞的增殖（P<0.05）。（2）二甲双胍和脂联素对EC细胞具有协同抗增殖作用，IK细胞的联合指数（CI）值为0.906 34，HEC-1B细胞的联合指数（CI）值为0.827 65。（3）运用qRT-PCR分别检测5 mmol/L和10 mmol/L二甲双胍作用IK和HEC-1B细胞48小时后AdipoR1和AdipoR2的mRNA水平，与对照组（0 mmol/L）相比，AdipoR1和AdipoR2的mRNA表达均显著增加（IK：5 mmol/L组和10 mmol/L组的AdipoR1：P<0.001，5 mmol/L组的AdipoR2：P<0.001；HEC-1B：5 mmol/L组的AdipoR1：P<0.001，10 mmol/L组：P=0.023，5 mmol/L组的AdipoR2：P<0.001，10 mmol/L组：P=0.024）。与化合物C联合使用时，AdipoR1和AdipoR2的RNA水平与对照组（0 mmol/L）相比无差异（P>0.05）。（4）用蛋白质免疫印迹法检测5 mmol/L和10 mmol/L二甲双胍作用IK和HEC-1B细胞48小时后AdipoR1和AdipoR2的蛋白水平，与对照组（0 mmol/L）相比，蛋白水平显著增加（IK：5 mmol/L组的AdipoR1：P=0.04，10 mmol/L组：P=0.033，5 mmol/L组的AdipoR2：P=0.044，10 mmol/L组：P=0.046；HEC-1B：5 mmol/L组的AdipoR1：P=0.从04，10 mmol/L组：P=0.049，5 mmol/L组的AdipoR2：P=0.043，10 mmol/L组：P=0.035）。与化合物C联合使用时，AdipoR1和AdipoR2的蛋白水平与对照组（0 mmol/L）相比无差异（P>0.055）。
二甲双胍和脂联素对EC细胞具有协同抗增殖作用。此外，二甲双胍可增加EC细胞脂联素受体在mRNA和蛋白水平的表达，且该作用是通过激活AMPK信号通路实现的。
