标题：MiD49在肝癌中的表达与促增殖作用研究

      作者：翟丽娜 1，蒋玮 2，吕卓敏 1，张庆 1，答秀维 1，张洪新 1    (空军军医大学唐都医院疼痛科 1、重症监护中心 2，陕西 西安 710038)

      卷次： 2018年29卷14期

      【摘要】 目的 探讨线粒体动力蛋白MiD49在肝癌中的表达及其促增殖作用。方法 ①用实时定量 PCR(qRT-PCR)、Western blot及免疫组化实验方法，分别检测MiD49在肝癌细胞株以及50例肝癌手术患者癌与癌旁组织中的表达；② siRNA下调MiD49表达后，分别用MTT与克隆形成实验检测MiD49在肝癌细胞增殖中的调控作用；③ siRNA下调MiD49表达后，用流式细胞术检测MiD49在肝癌细胞凋亡中的调控作用。结果 ①肝癌组织中MiD49表达的mRNA与蛋白水平均显著高于癌旁组织[mRNA水平：癌 vs癌旁=(0.38±0.28) vs (0.26±0.20)；蛋白水平：癌 vs癌旁=(5.67±0.47) vs (3.20±0.33)]，差异均有统计学意义(P<0.05)；肝癌细胞株中MiD49表达高于正常肝细胞[5种肝癌细胞 vs正常肝细胞=(2.42±0.23)、(4.57±0.35)、(2.53±0.22)、(3.30±0.44)、(3.11±0.32) vs (1.00±0.08)]，差异有统计学意义(P<0.05)。②利用 siRNA下调MiD49表达可显著抑制肝癌细胞增殖[siRNA vs si-MiD49#1 vs si-MiD49#2=(4.78±0.37) vs (3.07±0.28) vs (3.26±0.21)]，差异有统计学意义(P<0.05)；利用 siRNA下调MiD49表达可显著抑制肝癌细胞克隆形成[siRNA vs si-MiD49#1 vs si-MiD49#2=(449.00±34.00) vs (239.00±20.52) vs (223.66±21.73)]，差异有统计学意义(P<0.05)。③利用 siRNA下调MiD49表达可促进肝癌细胞凋亡[siRNA vs si-MiD49#1 vs si-MiD49#2=(3.90±0.40) vs (12.67±0.96) vs (13.17±1.01)]，差异有统计学意义(P<0.05)。结论 线粒体动力蛋白MiD49在肝癌中表达显著上调，MiD49高表达可促进肝癌细胞增殖而抑制凋亡，提示MID49是潜在的促癌因子并可作为潜在的肿瘤治疗靶标。
      【关键词】 MiD49；细胞增殖；细胞凋亡；肝癌
      【中图分类号】 R735.7 【文献标识码】 A 【文章编号】 1003—6350（2018）14—1925—06

Experimental study on the expression and pro-proliferative function of MiD49 in hepatocellular carcinoma.
ZHAILi-na 1, JIANG Wei 2, LV Zhuo-min 1, ZHANG Qing 1, DA Xiu-wei 1, ZHANG Hong-xin 1. Department of Pain Treatment 1,Department of Intensive Care Unit 2, Tangdu Hospital, the Air Force Military Medical university, Xi’an 710038, Shaanxi, CHINA
【Abstract】 Objective To evaluate the expression of mitochondrial dynamics protein 49 (MiD49) and itspro-proliferative function in hepatocellular carcinoma (HCC). Methods ① qRT-PCR, western blotting and immunohis-tochemistry analysis were used to detect the expression levels of MiD49 in HCC cell lines and tumor and peritumor tis-sues of 50 patients with HCC. ② The effect of MiD49 knockdown on the proliferation of HCC cells was analyzed byMTT and colony formation assays. ③ The effect of MiD49 knockdown on the apoptosis of HCC cells was analyzed byflow cytometry. Results ① The expression of MiD49 mRNA and protein were higher in tumor tissues of HCC whencompared with peritumor tissues: (0.38±0.28) vs (0.26±0.20) for mRNA (P<0.05); (5.67±0.47) vs (3.20±0.33) for protein(P<0.05). The expression of MiD49 was higher in five kinds of HCC cell lines when compared with normal liver cellline: (2.42±0.23), (4.57±0.35), (2.53±0.22), (3.30±0.44), (3.11±0.32) vs (1.00±0.08), P<0.05.② Knockdown of MiD49significantly inhibited the proliferation of HCC cells: siRNA vs si-MiD49#1 vs si-MiD49#2=(4.78±0.37) vs (3.07±0.28)vs (3.26 ± 0.21), P<0.05. Knockdown of MiD49 significantly inhibited the colony formation of HCC cells: siRNA vssi-MiD49#1 vs si-MiD49#2=(449.00±34.00) vs (239.00±20.52) vs (223.66±21.73), P<0.05. ③ Knockdown of MiD49significantly induced the apoptosis of HCC cells: siRNA vs si-MiD49#1 vs si-MiD49#2=(3.90±0.40) vs (12.67±0.96) vs(13.17±1.01), P<0.05. Conclusion MiD49 is overexpressed in HCC, which accelerates cell proliferation and colonyformation, and suppresses the apoptosis of HCC cells, indicating that MiD49 acts as a potential oncogene and may serveas a drug target in HCC treatment.
      【Key words】 Mitochondrial dynamics protein; Cell proliferation; Apoptosis; Hepatocellular carcinoma·论 著·基金项目：国家自然科学基金面上项目(编号：81572304、81600478、81772934)