miR-183及Akt1蛋白在子宫内膜癌组织中的表达及临床意义

摘要

目的 探讨miR-183及Akt1蛋白在子宫内膜癌组织中的表达及临床意义.方法 收集2014年3月至2016年9月暨南大学附属第一医院手术切除的60例子宫内膜癌标本及癌旁组织标本,术后均经病理学确诊.培养子宫内膜癌Ishikawa细胞株,采用LipofectamineTM2000脂质体转染试剂进行转染,将其分为A组(转染miR-183的特异性抑制剂反义寡核苷酸)、B组(转染无关miR-183的特异性抑制剂核苷酸序列)、C组(转染LipofectamineTM2000脂质体转染试剂)、D组(空白对照,不转染).采用流式细胞仪检测四组子宫内膜癌Ishikawa细胞的凋亡情况,实时逆转录聚合酶链反应检测各组子宫内膜癌Ishikawa细胞中miR-183及Akt1mRNA表达水平,蛋白质印迹法检测各组子宫内膜癌Ishikawa细胞中Akt1蛋白的表达水平.结果 子宫内膜癌组织中miR-183表达水平低于癌旁组织,Akt1蛋白mRNA表达水平高于癌旁组织(P<0.05).A组子宫内膜癌Ishikawa细胞早期凋亡率和晚期凋亡率均显著低于B、C、D组(P<0.05),B组早期凋亡率和晚期凋亡率均显著高于A、C、D组(P<0.05).A组子宫内膜癌Ishikawa细胞中miR-183表达水平低于B、C、D组(P<0.05).A、B组子宫内膜癌Ishikawa细胞中Akt1mRNA表达水平均显著低于C、D组(P<0.05).A组Akt1蛋白表达水平显著高于B、C、D组(P<0.05),B组Akt1蛋白表达水平显著低于C、D组(P<0.05).结论 miR-183在子宫内膜癌组织中呈低表达, Akt1蛋白呈高表达;在子宫内膜癌Ishikawa细胞中miR-183通过抑制Akt1蛋白的表达影响癌细胞增殖和凋亡,进而影响子宫内膜癌的发生和发展.%Objective To investigate the expression and clinical significance of miR-183 and Akt1 protein in endometrial carcinoma. Method From March 2014 to September 2016, 60 specimens of endometrial carcinoma and adjacent non-cancerous tissues specimens removed in first affiliated hospital of jinan university were collected, and all specimens were confirmed by pathology after operation. Cultured endometrial carcinoma cell lines Ishikawa, LipofectamineTM2000 were transfected by liposome transfection reagent, which were divided into group A (transfected with miR-183 antisense oligonucleotide specific inhibitor), group B (a specific inhibitor of the nucleotide sequence of miR-183 C, group C (transfected LipofectamineTM2000 liposome transfection reagent) and group D (blank control, notransfection). The apoptosis of four groups of endometrial carcinoma Ishikawa cells was detected by flow cytometry. The expression levels of miR-183 and Akt1 mRNA in endometrial carcinoma Ishikawa cells were detected by rea-time reverse transcription polymerase chain reaction. The expression level of Akt1 protein in endometrial carcinoma Ishikawa cells was detected by Western blotting. Result The expression of miR-183 in endometrial carcinoma was lower than that of adjacent non-cancerous tissues, and the expression of Akt1 mRNA was higher than that of adjacent non-cancerous tissues (P<0.05). The early apoptosis rate and late apoptosis rate in group A were lower than that in group B, C and D (P < 0.05), of which group B was the highest (P<0.05). The expression of miR-183 in Ishikawa cells of endometrial carcinoma of group A was lower than that of group B, C and D (P<0.05). The expression of Akt1 mRNA in Ishikawa cells of endometrial carcinoma of group A and group B was lower than that of group C and D (P< 0.05). The expression of Akt1 protein of group A was higher than that of group B, C and D (P<0.05),and group B was lower than that of group C and D (P < 0.05). Conclusion The expression of miR-183 was low in endometrial carcinoma and high expression of Akt1 protein. MiR-183 affects the proliferation and apoptosis of endometrial carcinoma Ishikawa cells by inhibiting the expression of Akt1 protein, and then affects the occurrence and development of endometrial carcinoma.