长链非编码RNA HAND2-AS1超声微泡通过调控miR-873-5p/基质金属蛋白酶-2组织抑制剂轴抑制肝细胞癌进展。
Long noncoding RNAs HAND2-AS1 ultrasound microbubbles suppress hepatocellular carcinoma progression by regulating the miR-873-5p/tissue inhibitor of matrix metalloproteinase-2 axis.
作者信息
Zou Qiang, Wang Hao-Wen, Di Xi-Liang, Li Yuan, Gao Hui
机构信息
Department of Interventional Therapy, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, China.
National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer, Tianjin 300060, China.
出版信息
World J Gastrointest Oncol. 2024 Apr 15;16(4):1547-1563. doi: 10.4251/wjgo.v16.i4.1547.
BACKGROUND
Increasing data indicated that long noncoding RNAs (lncRNAs) were directly or indirectly involved in the occurrence and development of tumors, including hepatocellular carcinoma (HCC). Recent studies had found that the expression of lncRNA HAND2-AS1 was downregulated in HCC tissues, but its role in HCC progression is unclear. Ultrasound targeted microbubble destruction mediated gene transfection is a new method to overexpress genes.
AIM
To study the role of ultrasound microbubbles (UTMBs) mediated HAND2-AS1 in the progression of HCC, in order to provide a new reference for the treatment of HCC.
METHODS
, we transfected HAND2-AS1 siRNA into HepG2 cells by UTMBs, and detected cell proliferation, apoptosis, invasion and epithelial-mesenchymal transition (EMT) by cell counting kit-8 assay, flow cytometry, Transwell invasion assay and Western blotting, respectively. In addition, we transfected miR-837-5p mimic into UTMBs treated cells and observed the changes of cell behavior. Next, the UTMBs treated HepG2 cells were transfected together with miR-837-5p mimic and tissue inhibitor of matrix metalloproteinase-2 (TIMP2) overexpression vector, and we detected cell proliferation, apoptosis, invasion and EMT. , we established a mouse model of subcutaneous transplantation of HepG2 cells and observed the effect of HAND2-AS1 silencing on tumor formation ability.
RESULTS
We found that UTMBs carrying HAND2-AS1 restricted cell proliferation, invasion, and EMT, encouraged apoptosis, and HAND2-AS1 silencing eliminated the effect of UTMBs. Additionally, miR-873-5p targets the gene HAND2-AS1, which also targets the 3'UTR of TIMP2. And miR-873-5p mimic counteracted the impact of HAND2-AS1. Further, miR-873-5p mimic solely or in combination with pcDNA-TIMP2 had been transformed into HepG2 cells exposed to UTMBs. We discovered that TIMP2 reversed the effect of miR-873-5p mimic caused by the blocked signalling cascade for matrix metalloproteinase (MMP) 2/MMP9. results showed that HAND2-AS1 silencing significantly inhibited tumor formation in mice.
CONCLUSION
LncRNA HAND2-AS1 promotes TIMP2 expression by targeting miR-873-5p to inhibit HepG2 cell growth and delay HCC progression.
背景
越来越多的数据表明，长链非编码RNA（lncRNAs）直接或间接参与肿瘤的发生和发展，包括肝细胞癌（HCC）。最近的研究发现，lncRNA HAND2-AS1在HCC组织中的表达下调，但其在HCC进展中的作用尚不清楚。超声靶向微泡破坏介导的基因转染是一种过表达基因的新方法。
目的
研究超声微泡（UTMBs）介导的HAND2-AS1在HCC进展中的作用，为HCC的治疗提供新的参考。
方法
我们通过UTMBs将HAND2-AS1 siRNA转染到HepG2细胞中，并分别通过细胞计数试剂盒-8检测、流式细胞术、Transwell侵袭试验和蛋白质印迹法检测细胞增殖、凋亡、侵袭和上皮-间质转化（EMT）。此外，我们将miR-837-5p模拟物转染到经UTMBs处理的细胞中，并观察细胞行为的变化。接下来，将经UTMBs处理的HepG2细胞与miR-837-5p模拟物和基质金属蛋白酶-2（TIMP2）过表达载体一起转染，我们检测细胞增殖、凋亡、侵袭和EMT。我们建立了HepG2细胞皮下移植小鼠模型，并观察HAND2-AS1沉默对肿瘤形成能力的影响。
结果
我们发现携带HAND2-AS1的UTMBs抑制细胞增殖、侵袭和EMT，促进凋亡，而HAND2-AS1沉默消除了UTMBs的作用。此外，miR-873-5p靶向基因HAND2-AS1，而HAND2-AS1也靶向TIMP2的3'UTR。并且miR-873-5p模拟物抵消了HAND2-AS1的影响。进一步地，miR-873-5p模拟物单独或与pcDNA-TIMP2组合转染到经UTMBs处理的HepG2细胞中。我们发现TIMP2逆转了miR-873-5p模拟物由基质金属蛋白酶（MMP）2/MMP9信号级联阻断引起的作用。结果表明，HAND2-AS1沉默显著抑制小鼠肿瘤形成。
结论
lncRNA HAND2-AS1通过靶向miR-873-5p促进TIMP2表达，从而抑制HepG2细胞生长并延缓HCC进展。
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