低氧/厌氧产品案例——低氧与肝癌研究

文章题目：Silencing of HIF-1α inhibited the expression of lncRNA NEAT1 to suppress development of hepatocellular carcinoma under hypoxia

沉默HIF-1α可以抑制lncRNA NEAT1 的表达，从而抑制缺氧条件下肝细胞癌的发展

文章出处：EAm J Transl Res 2020;12(7):3871-3883.中国南京医科大学附属肿瘤医院江苏省肿瘤医院放射科

工作站使用情况：Ruskinn Hypoxia Work Station

使用气体浓度：低氧（1% O₂）

摘要：本研究旨在探讨缺氧诱导因子-1α (HIF-1α)和lncRNA 核富集丰富转录本1 (NEAT1)之间的关系，以及它们在缺氧条件下肝细胞癌(HCC)中的作用。NEAT1 和HIF-1α在肝细胞癌组织中呈高表达并呈正相关，其中NEAT1 高表达与肿瘤淋巴结转移（TNM）晚期及远处转移呈正相关； NEAT1 或HIF-1α上调的HCC 患者预后较差。NEAT1由HIF-1α诱导，siHIF-1α抑制NEAT1 表达；NEAT1 的过表达进一步促进了缺氧条件下肝癌的发展，同时促进细胞活力、迁移和侵袭，抑制细胞凋亡，下调HIF-1α可逆转这种作用。NEAT1 过表达促进肿瘤生长，下调HIF-1α可逆转NEAT1 促进肿瘤生长的发生， 揭示下调HIF-1α可抑制NEAT1 的表达，抑制HCC 的进展，改善预后。

Figure 1. Correlations and relative expressions of HIF-1α and NEAT1 in HCC tissues and their relationships with overall survival of HCC patients. (A and B) HIF-1α (A) and NEAT1 (B) expressions were detected by qRT-PCR and normalized to GAPDH expression in 98 pairs of HCC tissues compared with adjacent nontumourous liver specimens. (C) Spearman’s rank correlation analysis was performed to analyze the correlations of HIF-1α and NEAT1 in HCC tissues. (D and E) Kaplan-Meier survival curve and log-rank test were used to evaluate whether HIF-1α and NEAT1 expression levels were associated with overall survival rate. ***P < 0.001 vs. Adjacent tissue. The data were shown as mean ± standard deviation (S.D.). Abbreviations: HIF-1α, hypoxia-inducible factors-1α; NEAT1, lncRNA nuclear enriched abundant transcript 1; qRT-PCR, quantitative reverse transcription-polymerase chain reaction; HCC, hepa tocellular carcinoma.

Figure 4. HIF-1α silencing suppressed the development of HCC by negatively regulating NEAT1 level in Hep3B and SK-Hep1 cells under hypoxia. (A, B) Migration rates were detected in Normal (non-transfected and non-hypoxia control), Hypoxia, pc-Control (non-targeting and hypoxia control), pc-NEAT1 (NEAT1 transfected and hypoxia), pc-NEAT1 + siHIF-1α (NEAT1 and siRNA-HIF-1α co-transfected under hypoxia), siHIF-1α groups in Hep3 (A and B) and SK-Hep1 cells (C and D) by scratch wound test. (E-H) Invasion rates were detected in Normal, Hypoxia, pc-Control, pc-NEAT1, pc-NEAT1 + siHIF-1α, siHIF-1α groups in Hep3 (E and F) and SK-Hep1 cells (G and H) by transwell assay. ***P < 0.001 vs. Normal. ###P < 0.001 vs. pc-Control. ^^^P < 0.001 vs. pc-NEAT1. ΔΔΔP < 0.001 vs. pc- NEAT1 + siHIF-1α. The data were shown as mean ± standard deviation (S.D.). Abbreviations: HIF- 1α, hypoxia-inducible factors-1α; NEAT1, lncRNA nuclear-enriched abundant transcript 1.

       HCC 组织中NEAT1 和HIF1α明显升高(图1A 和1B)；相关性分析显示HIF-1α的表达与NEAT1 的表达呈正相关（图1C），HIF-1α高表达组存活的HCC 患者比例明显低于HIF-1α低表达组（(图1D），提示HIF-1α低表达的HCC 患者预后较好；

       NEAT1 过表达进一步促进了缺氧条件下的迁移和侵袭，沉默HIF-1α显著抑制了缺氧条件下的迁移和侵袭，而NEAT1 过表达部分逆转了沉默HIF-1α的作用（图4A-H）；表明HIF-1α沉默通过负向调节缺氧条件下Hep3B 和SK-Hep1 细胞NEAT1 水平抑制肝癌的发生发展。