背景:据报道,长链非编码RNA转移相关的肺腺癌转录物1(MALAT1)在各种肿瘤的发生和发展中起着至关重要的作用。然而,MALAT1在肝细胞癌(HCC)中的潜在机制尚未完全阐明。
方法:采用qRT-PCR检测MALAT1在肝癌组织和不同细胞系中的表达水平。用反义寡核苷酸(ASO)-MALAT1转染细胞,通过细胞计数试剂盒8(CCK-8)探讨MALAT1在肝癌细胞中的生物学效应,菌落形成,transwell,伤口愈合,和流式细胞术分析。进行蛋白质印迹以测量AMPK和凋亡相关蛋白水平。进行双荧光素酶报告基因测定以验证MALAT1与其特异性靶标之间的关系。
结果:我们发现MALAT1在HCC中上调,和MALAT1敲低肝癌细胞抑制细胞增殖,迁移,和侵袭,并在体外抑制细胞凋亡。进一步的研究表明,MALAT1正调节转录因子IIB相关因子2(BRF2)的表达,这与肿瘤复发有关,大肿瘤大小,肝癌预后不良。机械上,MALAT1被发现作为海绵has-miR-1-3p的竞争性内源性RNA,这上调了BRF2的表达。BRF2的敲低通过激活LKB1/AMPK信号通路抑制HCC的进展。BRF2的过表达逆转了MALAT1敲低对HCC细胞活力的抑制作用。此外,靶向MALAT1的ASO抑制异种移植肿瘤的生长。
结论:我们的结果证明了HCC中一种新的MALAT1/miR-1-3p/BRF2/LKB1/AMPK调节轴,这可能为未来HCC提供新的分子治疗靶点。
BACKGROUND: The long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) has been reported to play a vital role in the occurrence and development of various tumors. However, the underlying mechanism of MALAT1 in hepatocellular carcinoma (HCC) has not been thoroughly elucidated.
METHODS: The expression levels of MALAT1 in HCC tissues and different cell lines were detected by qRT-PCR. Antisense oligonucleotides (ASO)-MALAT1 transfected cells were used to explore the biological effects of MALAT1 in HCC cells by cell counting kit 8 (CCK-8), colony formation, transwell, wound healing, and flow cytometry analysis. Western blotting was performed to measure AMPK and apoptosis-related protein levels. Dual-luciferase reporter assay was performed to verify the relationship between MALAT1 and its specific targets.
RESULTS: We found that MALAT1 was upregulated in HCC, and MALAT1 knockdown in HCC cells inhibited cell proliferation, migration, and invasion and inhibited apoptosis in vitro. Further studies demonstrated that MALAT1 positively regulated the expression of transcription factor II B‑related factor 2 (
BRF2), which was associated with tumor recurrence, large tumor size, and poor prognosis in HCC. Mechanistically, MALAT1 was found to act as a competitive endogenous RNA to sponge has-miR-1-3p, which upregulated
BRF2 expression. Knockdown of
BRF2 inhibited the progression of HCC by activating the LKB1/AMPK signaling pathway. Overexpression of
BRF2 reversed the inhibitory effect of MALAT1 knockdown on HCC cell viability. Moreover, ASO targeting MALAT1 inhibited the growth of xenograft tumors.
CONCLUSIONS: Our results demonstrate a novel MALAT1/miR-1-3p/
BRF2/LKB1/AMPK regulatory axis in HCC, which may provide new molecular therapeutic targets for HCC in the future.