lncRNA TSIX通过靶向miR-548-a-3p抑制肝细胞癌细胞增殖和侵袭的机制研究

肝脏 ›› 2022, Vol. 27 ›› Issue (2): 213-217.

lncRNA TSIX通过靶向miR-548-a-3p抑制肝细胞癌细胞增殖和侵袭的机制研究

王佳乐, 李琤, 李广明

450000 郑州河南省传染病医院(郑州市第六人民医院)肝硬化科

收稿日期:2021-05-22 出版日期:2022-02-28 发布日期:2022-04-19
通讯作者: 李广明
基金资助:
河南省医学科技攻关计划项目(2018020849)

The mechanism of lncRNA TSIX inhibiting proliferation and invasion of hepatocellular carcinoma cell by targeting miR-548-a-3p

WANG Jia-le, LI Cheng, LI Guang-ming

Department of Liver Cirrhosis, Henan Provincial Hospital of Infectious Diseases, Henan 450000, China

Received:2021-05-22 Online:2022-02-28 Published:2022-04-19
Contact: LI Guang-ming

摘要/Abstract

摘要： 目的探讨lncRNA TSIX通过靶向miR-548-a-3p抑制肝细胞癌(HCC)细胞增殖和侵袭的机制。方法设肝癌Huh7细胞组、lncRNA TSIX mimics组、lncRNA TSIX inhibitor组;各组细胞培养48 h后,采用MTT法测定细胞活力,结晶紫染色测定单克隆形成数目,流式细胞仪测定细胞凋亡水平,transwell腔室测定细胞侵袭水平,RT-PCR法测定细胞lncRNA TSIX、miR-548-a-3p水平。结果 lncRNA TSIX mimics组OD值、存活率、单克隆形成数目、穿膜数低于Huh7细胞组(t=8.725、5.437、120.834、64.321,P<0.05),lncRNA TSIX inhibitor组OD值、存活率、单克隆形成数目、穿膜数高于Huh7细胞组和lncRNA TSIX mimics组(t=4.124、3.784、10.462、7.965、37.192、174.632、30.703、104.562,P<0.05)。lncRNA TSIX mimics组细胞凋亡率、lncRNA TSIX高于Huh7细胞组[(18.52±2.45)vs(15.63±2.13),(4.47±0.22)vs(2.89±0.21)](t=10.531、12.372,P<0.05),lncRNA TSIX inhibitor组细胞凋亡率、lncRNA TSIX低于Huh7细胞组和lncRNA TSIX mimics组[(8.14±2.45)vs(15.63±2.13)、(18.52±2.45),(1.58±0.25)vs(2.89±0.21)、(4.47±0.22)](t=37.385、48.021、30.703、104.562,P<0.05)。lncRNA TSIX mimics组miR-548-a-3p低于Huh7细胞组[(2.39±0.19) vs (4.87±0.20)](t=11.265,P<0.05),lncRNA TSIX inhibitor组miR-548-a-3p高于Huh7细胞组和lncRNA TSIX mimics组[(6.32±0.19)vs(4.87±0.20)、(2.39±0.19)](t=6.612、10.058,P<0.05)。结论 lncRNA TSIX明显抑制HCC细胞增殖、侵袭,促进HCC细胞凋亡;其机制可能与lncRNA TSIX负调节miR-548-a-3p有关。

关键词: lncRNA TSIX, miR-548-a-3p, 肝细胞癌, 细胞增殖, 细胞侵袭

Abstract: Objective To investigate the mechanism of lncRNA TSIX inhibiting proliferation and invasion of hepatocellular carcinoma cell by targeting miR-548-a-3p. Methods The liver cancer cell line Huh7 group, lncRNA TSIX mimics group, and lncRNA TSIX inhibitor group were set up. After 48 hours of cell culturing in each group, the cell viability was measured by MTT method, and the number of single clones formed by crystal violet staining was measured, and the apoptosis level was measured by flow cytometry. The cell invasion level was measured by transwell chamber, and the lncRNA TSIX and miR-548-a-3p levels were measured by RT-PCR method. Results The OD value, survival rate, number of colonies formed, and number of membrane penetration in the lncRNA TSIX mimics group were lower than those in the Huh7 group (t=8.725, 5.437, 120.834, 64.321, P<0.05), and the OD value, survival rate, number of colonies formed, number of membrane penetration in the lncRNA TSIX inhibitor group were higher than those in the Huh7 group and lncRNA TSIX inhibitor group (t=4.124, 3.784, 10.462, 7.965, 37.192, 174.632, 30.703, 104.562, P<0.05). The apoptotic rate and lncRNA TSIX in the lncRNA TSIX mimics group was higher than those in the Huh7 group [(18.52±2.45) vs (15.63±2.13), (4.47±0.22) vs (2.89±0.21)] (t=10.531, 12.372, P<0.05), and the apoptotic rate and lncRNA TSIX in the lncRNA TSIX inhibitor group was lower than those in the Huh7 group and the lncRNA TSIX inhibitor group [(8.14±2.45) vs (15.63±2.13), (18.52±2.45); (1.58±0.25) vs (2.89±0.21), (4.47±0.22)] (t=37.385, 48.021, 30.703, 104.562, P<0.05). The miR-548-a-3p in lncRNA TSIX mimics group was lower than that in Huh7 cell group [(2.39±0.19) vs (4.87±0.20)] (t=11.265, P<0.05), and miR-548-a-3p in lncRNA TSIX inhibitor group was higher than that in Huh7 group and lncRNA TSIX inhibitor group [(6.32±0.19) vs (4.87±0.20), (2.39±0.19)] (t=6.612, 10.058, P<0.05). Conclusion LncRNA Tsix significantly inhibited the proliferation and invasion of HCC cells and promoted the apoptosis of HCC cells. The mechanism may be related to the negative regulation of miR-548-a-3p by lncRNA Tsix.

Key words: LncRNA TSIX, MiR-548-a-3p, Hepatocellular carcinoma, Cell proliferation, Cell invasion

王佳乐, 李琤, 李广明. lncRNA TSIX通过靶向miR-548-a-3p抑制肝细胞癌细胞增殖和侵袭的机制研究[J]. 肝脏, 2022, 27(2): 213-217.

WANG Jia-le, LI Cheng, LI Guang-ming. The mechanism of lncRNA TSIX inhibiting proliferation and invasion of hepatocellular carcinoma cell by targeting miR-548-a-3p[J]. Chinese Hepatolgy, 2022, 27(2): 213-217.

参考文献

[1] Yang JD, Hainaut P, Gores GJ, et al. A global view of hepatocellular carcinoma: trends, risk, prevention and management. Nat Rev Gastroenterol Hepatol, 2019, 16: 589-604.
[2] Kulik L, El-Serag HB. Epidemiology and management of hepatocellular carcinoma. Gastroenterology, 2019, 156: 477-491. e1.
[3] Jiang MC, Ni JJ, Cui WY, et al. Emerging roles of lncRNA in cancer and therapeutic opportunities. Am J Cancer Res, 2019, 9: 1354-1366.
[4] Yang X, Zhang S, He C, et al. METTL14 suppresses proliferation and metastasis of colorectal cancer by down-regulating oncogenic long non-coding RNA XIST. Mol Cancer, 2020, 19: 46.
[5] Habieb A, Matboli M, El-Tayeb H, et al. Potential role of lncRNA-TSIX, miR-548-a-3p, and SOGA1 mRNA in the diagnosis of hepatocellular carcinoma. Mol Biol Rep, 2019, 46: 4581-4590.
[6] Bu Y, Zheng D, Wang L, et al. LncRNA TSIX promotes osteoblast apoptosis in particle-induced osteolysis by down-regulating miR-30a-5p. Connect Tissue Res, 2018, 59: 534-541.
[7] Kim J, Piao HL, Kim BJ, et al. Long noncoding RNA MALAT1 suppresses breast cancer metastasis. Nat Genet, 2018, 50: 1705-1715.
[8] Zhu H, Zheng T, Yu J, et al. LncRNA XIST accelerates cervical cancer progression via upregulating Fus through competitively binding with miR-200a. Biomed Pharmacother, 2018, 105: 789-797.
[9] Strehle M, Guttman M. Xist drives spatial compartmentalization of DNA and protein to orchestrate initiation and maintenance of X inactivation. Curr Opin Cell Biol, 2020, 64: 139-147.
[10] Yang C, Wu K, Wang S, et al. Long non-coding RNA XIST promotes osteosarcoma progression by targeting YAP via miR‐195‐5p. J Cell Biochem, 2018, 119: 5646-5656.
[11] Li C, Wan L, Liu Z, et al. Long non-coding RNA XIST promotes TGF-β-induced epithelial-mesenchymal transition by regulating miR-367/141-ZEB2 axis in non-small-cell lung cancer. Cancer Lett, 2018, 418: 185-195.
[12] Saffari M, Ghaderian SMH, Omrani MD, et al. The Association of miR-let 7b and miR-548 with PTEN in Prostate Cancer. Urology J, 2019, 16: 267-273.
[13] Yang C, Li Q, Chen X, et al. Circular RNA circRGNEF promotes bladder cancer progression via miR-548/KIF2C axis regulation. Aging (Albany NY), 2020, 12: 6865-6879.
[14] Salah SMM, Matboli M, Nasser HE, et al. Dysregulation in the expression of (lncRNA-TSIX, TP53INP2 mRNA, miRNA-1283) in spinal cord injury. Genomics, 2020, 112:3315-3321.
[15] Xu WW, Xu Y, Ji F, et al. Inhibition of long non-coding RNA TSIX accelerates tibia fraction healing via binding and positively regulating the SOX6 expression. Eur Rev Med Pharmacol Sci, 2020, 24:4070-4079.