Du Shuixian, Liao Songling, Liu Shousheng, Xin Yongning
Department of Infectious Disease, Qingdao Municipal Hospital, Qingdao, China.
Department of Gastroenterology, Dalian Medical University, Dalian, China.
J Clin Transl Hepatol. 2019 Mar 28;7(1):27-31. doi: 10.14218/JCTH.2018.00055. Epub 2019 Feb 8.
Accumulated evidence has shown that chronic liver inflammation is one of the main risks of hepatocellular carcinoma (HCC), and E167K variant of the transmembrane 6 superfamily member 2 (TM6SF2) plays an important role in the progression of chronic liver diseases and HCC. The aim of this study was to explore effects of the TM6SF2 E167K variant on expression of the inflammatory cytokines TNF-α, IL-2, IL-6 and IL-8 in the HCC cell line HEPA 1-6. HEPA 1-6 cells were infected with lentivirus containing either the TM6SF2 E167K variant or TM6SF2 wild-type, or control plasmids. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were conducted to analyze the expression of the inflammatory cytokines TNF-α, IL-2, IL-6 and IL-8. A t-test was used for statistical analysis. Compared with the control group and TM6SF2 overexpression group, the relative expression of IL-2 and IL-6 mRNAs were significantly elevated in the TM6SF2 E167K overexpression group ( < 0.05). The relative mRNA expression of IL-8 in the TM6SF2 and TM6SF2 E167K overexpression groups were increased compared to the control group ( < 0.05). No obvious differences were observed for the expression of TNF-α in each group. The expression of TNF-α, IL-2, IL-6 and IL-8 that was tested by western blotting showed the same trends as the qRT-PCR results. In conclusion, the E167K variant of the TM6SF2 gene could promote the expression of inflammatory cytokines IL-2 and IL-6 in HEPA 1-6 cells, suggesting that the TM6SF2 E167K variant may accelerate the progression of HCC.
越来越多的证据表明，慢性肝脏炎症是肝细胞癌（HCC）的主要风险之一，跨膜6超家族成员2（TM6SF2）的E167K变体在慢性肝病和HCC的进展中起重要作用。本研究的目的是探讨TM6SF2 E167K变体对肝癌细胞系HEPA 1-6中炎性细胞因子TNF-α、IL-2、IL-6和IL-8表达的影响。用含有TM6SF2 E167K变体或TM6SF2野生型的慢病毒或对照质粒感染HEPA 1-6细胞。采用定量实时聚合酶链反应（qRT-PCR）和蛋白质印迹法分析炎性细胞因子TNF-α、IL-2、IL-6和IL-8的表达。采用t检验进行统计学分析。与对照组和TM6SF2过表达组相比，TM6SF2 E167K过表达组中IL-2和IL-6 mRNA的相对表达显著升高（<0.05）。与对照组相比（<0.05），TM6SF2和TM6SF2 E167K过表达组中IL-8的相对mRNA表达增加。各组TNF-α的表达未观察到明显差异。蛋白质印迹法检测的TNF-α、IL-2、IL-6和IL-8的表达与qRT-PCR结果显示出相同趋势。总之，TM6SF2基因的E167K变体可促进HEPA 1-6细胞中炎性细胞因子IL-2和IL-6的表达，提示TM6SF2 E167K变体可能加速HCC的进展。
