MFF在肝癌中的表达及其生物学作用

doi:10.3760/cma.j.issn.1673-422X.2018.01.004

摘要/Abstract

摘要： 目的探讨线粒体分裂因子(MFF)在肝癌中的表达与生物学作用。方法①用实时荧光定量PCR（qPCR）、Western blotting及免疫组织化学方法，检测MFF在肝癌组织与细胞株中的表达。②小干扰RNA（siRNA）干涉MFF表达后，用四甲基偶氮唑蓝法(MTT)与克隆形成实验检测siCtrl、siMFF#1、siMFF#2组MFF对肝癌细胞增殖的影响。③siRNA干涉MFF表达后，用Annexin ⅤFITC/PI双染法检测MFF对肝癌细胞凋亡的影响。结果①MFF在肝癌组织中的表达显著高于癌旁组织［mRNA水平M（QR）：0.292（0.443）∶0.235（0.333），Z=-4.166，P＜0.001；蛋白水平M（QR）：5.414（4.545）∶3.120（3.955），Z=-3.961，P＜0.001］；MFF在所有被检测肝癌细胞株中的表达均高于正常肝细胞。②干涉MFF可显著抑制肝癌细胞增殖（siCtrl vs. siMFF#1：5.29±0.34∶3.34±0.37，P=0.014；siCtrl vs. siMFF#2：5.29±0.34∶3.09±0.40，P=0.010）；干涉MFF可显著抑制肝癌细胞克隆形成（siCtrl vs. siMFF#1：95.35±21.20∶37.56±10.61，P=0.003；siCtrl vs. siMFF#2：95.35±21.20∶41.23±10.82，P=0.004）。③干涉MFF可明显促进肝癌细胞凋亡（siCtrl vs. siMFF#1：9.56%±1.70%∶20.08%±2.03%，P＜0.001；siCtrl vs. siMFF#2：9.56%±1.70%∶21.14%±1.38%，P＜0.001）。结论MFF在肝癌中高表达，可促进肝癌细胞增殖并抑制凋亡，提示MFF是潜在的癌基因与肿瘤治疗靶标。

关键词: 肝肿瘤, 细胞增殖, 细胞凋亡, 线粒体分裂因子

Abstract: ObjectiveTo evaluate the expression of mitochondrial fission factor (MFF) and its biological effects in the progression of hepatocellular carcinoma (HCC). Methods①Quantitative realtime PCR (qPCR), Western blotting and immunohistochemistry analysis were used to detect the expression levels of MFF in HCC tumor tissues and cell lines. ②The effect of MFF knockdown on proliferation of HCC cells was analyzed by methyl thiazolyl tetrazolium (MTT) and colony formation assays in siCtrl, siMFF#1, siMFF#2 groups. ③The effect of MFF knockdown on apoptosis of HCC cells was analyzed by apoptosis assay with Annexin ⅤFITC and PI. Results①The MFF expression was higher in tumor tissues compared with tumoradjacent normal tissues ［mRNA level M(QR): 0.292(0.443) vs. 0.235(0.333), Z=-4.166, P＜0.001; protein level M(QR): 5.414 (4.545) vs. 3.120 (3.955), Z=-3.961， P＜0.001)］. The MFF expression was higher in HCC cell lines compared with normal liver cell line. ②RNA interferencemediated knockdown of MFF inhibited proliferation of HCC cells (siCtrl vs. siMFF#1: 5.29±0.34 vs. 3.34±0.37, P=0.014; siCtrl vs. siMFF#2: 5.29±0.34 vs. 3.09±0.40, P=0.010). RNA interferencemediated knockdown of MFF inhibited colony formation of HCC cells (siCtrl vs. siMFF#1: 95.35±21.20 vs. 37.56±10.61, P=0.003; siCtrl vs. siMFF#2: 95.35±21.20 vs. 41.23±10.82, P=0.004). ③RNA interferencemediated knockdown of MFF induced apoptosis of HCC cells (siCtrl vs. siMFF#1: 9.56%±1.70% vs. 20.08%±2.03%, P＜0.001; siCtrl vs. siMFF#2: 9.56%±1.70% vs. 21.14%±1.38%, P＜0.001). ConclusionMFF is overexpressed in HCC, which accelerates cell proliferation and suppresses apoptosis, indicating that MFF can serve as a potential oncogene and drug target in HCC treatment.

Key words: Liver neoplasms, Cell proliferation, Apoptosis, Mitochondrial fission factor

李波,张建省,李嘉琦,杨懿,张洪新,牟佼. MFF在肝癌中的表达及其生物学作用[J]. 国际肿瘤学杂志, 2018, 45(1): 16-.

Li Bo*, Zhang Jiansheng, Li Jiaqi, Yang Yi, Zhang Hongxin, Mou Jiao. Expression of MFF and its biological effects in hepatocellular carcinoma[J]. Journal of International Oncology, 2018, 45(1): 16-.

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