脂联素对子宫内膜癌Ishikawa细胞增殖及侵袭和转移能力的影响

Effects of adiponectin on invasion and metastasis of endometrial carcinoma Ishikawa cells

目的探讨脂联素对子宫内膜癌Ishikawa细胞增殖、侵袭和转移的影响及其机制。方法将培养的子宫内膜癌Ishikawa细胞随机分为空白对照组、阴性对照组和实验组,取3组对数生长期Ishikawa细胞,采用脂质体转染法将脂联素反义RNA及其对照microRNA分别转入实验组及阴性对照组Ishikawa细胞,空白对照组细胞不进行转染。转染36 h后,采用四甲基偶氮唑盐法、克隆形成实验及Transwell实验检测Ishikawa细胞生长、增殖和迁移能力,采用western blot法检测Ishikawa细胞中脂联素蛋白及细胞外调节蛋白激酶(ERK)蛋白表达,采用流式细胞术检测Ishikawa细胞凋亡情况,应用caspase-3活性检测试剂盒检测Ishikawa细胞中caspase-3活性。结果实验组Ishikawa细胞增殖及侵袭能力大于空白对照组和阴性对照组(P<0.01),空白对照组与阴性对照组Ishikawa细胞增殖能力及侵袭能力比较差异无统计学意义(P>0.05)。实验组Ishikawa细胞膜表面磷脂酰丝氨酸表达及caspase-3活性低于空白对照组和阴性对照组(P<0.05),空白对照组与阴性对照组Ishikawa细胞膜表面磷脂酰丝氨酸表达及caspase-3活性比较差异无统计学意义(P>0.05)。实验组Ishikawa细胞中脂联素蛋白及ERK蛋白表达低于空白对照组和阴性对照组(P<0.01),空白对照组与阴性对照组Ishikawa细胞中脂联素蛋白及ERK蛋白表达比较差异无统计学意义(P>0.05)。实验组Ishikawa细胞克隆能力大于空白对照组和阴性对照组(P<0.05),空白对照组与阴性对照组Ishikawa细胞克隆能力能力比较差异无统计学意义(P>0.05)。结论脂联素可能通过对ERK信号通路的影响诱导子宫内膜癌细胞凋亡、抑制子宫内膜癌细胞侵袭和迁移而发挥抗肿瘤作用。

Objective To investigate the effect of adiponectin on proliferation,invasion and metastasis of endometrial carcinoma Ishikawa cells and its mechanism.Methods The cultured endometrial cancer Ishikawa cells were randomly divided into blank control group,negative control group and experimental group.The Ishikawa cells in the logarithmic growth phase in the three groups were collected.Adiponectin antisense RNA and its control microRNA were transfected into the Ishikawa cells in the experimental group and the negative control group by liposome transfection respectively,and the Ishikawa cells in the blank control group were not transfected.After transfection of 36 h,The growth,proliferation and migration ability of Ishikawa cells were detected by mthyl thiazolyl tetrazolium method,colony formation assay and Transwell assay;the expression of adiponectin and extracellular regulated protein kinase(ERK)protein in Ishikawa cells was detected by western blot method,the apoptosis of Ishikawa cells was detected by flow cytometry,and the activity of caspase-3 in Ishikawa cells was detected by caspase-3 activity assay kit.Results The proliferation and invasion ability of Ishikawa cells in the experimental group were greater than those in the blank control group and the negative control group(P<0.01).There was no significant difference in Ishikawa cell proliferation and invasion ability between the blank control group and negative control group(P>0.05).The expression of phosphatidylserine on the surface of Ishikawa cell membrane and the caspase-3 activity in Ishikawa cells in the experimental group were lower than those in the blank control group and negative control group(P<0.05),but there was no significant difference in the expression of phosphatidylserine and the caspase-3 activity in Ishikawa cells between the blank control group and negative control group(P>0.05).The expression of adiponectin and ERK protein in the Ishikawa cells in the experimental group was lower than that in the blank control group and the negative control group(P<0.01),but there was no significant difference in the expression of adiponectin and ERK protein in the Ishikawa cells between the blank control group and negative control group(P>0.05).The Ishikawa cell cloning ability in the experimental group was more than that in the blank control group and the negative control group(P<0.05),but there was no significant difference in the Ishikawa cell cloning ability between the blank control group and negative control group(P>0.05).Conclusion Adiponectin may play an anti-tumor role by inducing endometrial carcinoma cell apoptosis and inhibiting the invasion and migration of endometrial cancer cells by effecting ERK signaling pathway.