环状RNA_0003028靶向微小RNA-498调控生殖器形成抑制基因-1/p53 信号通路对人肝癌细胞系Huh7的影响

谢宇端; 董国伟; 韩艳

doi:10.16098/j.issn.0529-1356.2022.05.009

PDF(10338 KB)

解剖学报 ›› 2022, Vol. 53 ›› Issue (5) : 600-606. DOI: 10.16098/j.issn.0529-1356.2022.05.009

环状RNA_0003028靶向微小RNA-498调控生殖器形成抑制基因-1/p53 信号通路对人肝癌细胞系Huh7的影响

谢宇端^1,2 董国伟^2*韩艳² 何生奇³

作者信息 +

Circular RNA_0003028 targeting microRNA-498 regulating the effects of suppressor with morphogenesis effect on genitalia-1/p53 signaling pathway on liver cancer cell line Huh7

XIE Yu-duan^1,2 DONG Guo-wei^2* HAN Yan² HE Sheng-qi³

Author information +

文章历史 +

摘要

目的探讨环状RNA（circ）_0003028对人肝癌细胞增殖、迁移和侵袭的影响及分子机制。方法人肝癌细胞系Huh7分为小干扰RNA（si）-NC组、si-circ_0003028组、微小RNA（miR）-NC组、miR-498模似物（mimics）组、si-circ_0003028+anti-miR-NC组、si-circ_0003028+anti-miR-498组；Real\|time PCR检测肝癌组织及各组细胞中circ_0003028和miR-498表达水平；MTT检测细胞增殖；Transwell检测细胞迁移和侵袭数；Western blotting检测蛋白表达；双荧光素酶报告实验检测circ_0003028和miR-498的靶向调控关系。结果肝癌组织中circ_0003028表达水平升高（0.98±0.02 vs 1.36±0.01），miR-498表达水平降低（0.98±0.02 vs 0.63±0.02）（P<0.05）。抑制circ_0003028表达或miR-498过表达后，Huh7细胞中Ki-67（0.85±0.02 vs 0.41±0.02或0.95±0.11 vs 0.37±0.02）、基质金属蛋白酶（MMP）-2（0.71±0.02 vs 0.43±0.03或0.83±0.02 vs 0.41±0.03）、MMP-9（0.74±0.02 vs 0.37±0.02或0.78±0.02 vs 0.39±0.02）蛋白表达水平降低，细胞活性（1.53±0.03 vs 1.05±0.02或1.68±0.02 vs 1.11±0.02）降低；迁移（111.40±2.12 vs 77.22±2.38或108.90±2.30 vs 78.44±1.46）和侵袭（87.89±2.18 vs 49.78±1.98或80.22±1.79 vs 38.22±1.52）细胞数减少，生殖器形成抑制基因-1(SMG-1)（0.76±0.02 vs 1.39±0.02或0.79±0.02 vs 1.39±0.02）、p53（0.77±0.02 vs 1.24±0.03或0.82±0.03 vs 1.45±0.03）、p53-ser15（0.78±0.03 vs 1.50±0.02或0.82±0.02 vs 1.49±0.04）蛋白表达水平升高（P<0.05）。Circ_0003028靶向调控miR-498，沉默miR-498逆转了抑制circ_0003028表达对Huh7细胞增殖、迁移和侵袭的影响。结论抑制circ_0003028表达通过靶向miR-498影响SMG-1/p53 信号通路而抑制人肝癌细胞增殖、迁移和侵袭。

Abstract

Objective To investigate the effects and molecular mechanism of circular RNA(circ)_0003028 on proliferation, migration and invasion of human liver cancer cells. Methods Liver cancer cell line Huh7 were divided into small interfering RNA(si)-NC group, si-circ_0003028 group, microRNA(miR)-NC group, miR-498 mimics group, si-circ_0003028+anti-miR-NC group and si-circ_0003028+anti-miR-498 group. Real-time PCR was used to detect the expression levels of circ_0003028 and miR-498 in liver cancer tissues and cells of each groups. MTT was used to detect cell proliferation. Transwell was used to detect cell migration and invasion. Western blotting was used to detect protein expression. Dual luciferase reporter experiment was used to detect the target regulation relationship between circ_0003028 and miR-498. Results The expression level of circ_0003028(0.98±0.02 vs 1.36±0.01) increased and the expression level of miR-498(0.98±0.02 vs 0.63±0.02) decreased in liver cancer tissues (P<0.05). After inhibiting the expression of circ_0003028 or overexpression of miR-498, the expression levels of Ki-67(0.85±0.02 vs 0.41±0.02 or 0.95±0.11 vs 0.37±0.02), matrix metalloprotein(MMP)-2(0.71±0.02 vs 0.43±0.03 or 0.83±0.02 vs 0.41±0.03), and MMP-9(0.74±0.02 vs 0.37±0.02 or 0.78±0.02 vs 0.39±0.02) proteins in Huh7 cells decreased, and cell viability(1.53±0.03 vs 1.05±0.02 or 1.68±0.02 vs 1.11±0.02) decreased; the number of migration(111.40±2.12 vs 77.22±2.38 or 108.90±2.30 vs 78.44±1.46) and invasion(87.89±2.18 vs 49.78±1.98 or 80.22±1.79 vs 38.22±1.52) cells decreased, and the protein expression levels of suppressor with morphogenesis effect on genitalia-1(SMG-1) (0.76±0.02 vs 1.39±0.02 or 0.79±0.02 vs 1.39±0.02), p53(0.77±0.02 vs 1.24±0.03 or 0.82±0.03 vs 1.45±0.03), and p53-ser15(0.78±0.03 vs 1.50±0.02 or 0.82±0.02 vs1.4.04) increased (P<0.05). circ_0003028 targeted regulation of miR-498, and silencing miR-498 reversed the effects of inhibiting the expression of circ_0003028 on the proliferation, migration and invasion of Huh7 cells. Conclusion Inhibiting the expression of circ_0003028 can inhibit the proliferation, migration and invasion of liver cancer cells by targeting miR-498 to affect the SMG-1/p53 signaling pathway.

导出引用

谢宇端董国伟韩艳何生奇. 环状RNA_0003028靶向微小RNA-498调控生殖器形成抑制基因-1/p53 信号通路对人肝癌细胞系Huh7的影响[J]. 解剖学报. 2022, 53(5): 600-606 https://doi.org/10.16098/j.issn.0529-1356.2022.05.009

XIE Yu-duan DONG Guo-wei HAN Yan HE Sheng-qi. Circular RNA_0003028 targeting microRNA-498 regulating the effects of suppressor with morphogenesis effect on genitalia-1/p53 signaling pathway on liver cancer cell line Huh7[J]. Acta Anatomica Sinica. 2022, 53(5): 600-606 https://doi.org/10.16098/j.issn.0529-1356.2022.05.009

中图分类号： R735.7

参考文献

［1］Fu J, Wang H. Precision diagnosis and treatment of liver cancer in China［J］. Cancer Lett, 2018,412(1): 283-288.

［2］Kudo M. Targeted and immune therapies for hepatocellular carcinoma: Predictions for 2019 and beyond［J］. World J Gastroenterol, 2019,25(7): 789-807.

［3］Deng XJ, Gong ShX, Yang ShH, et al. MicroRNA-539 inhibiting the occurrence and development of hepatocellular carcinoma by targeting E2F transcription factor 3［J］. Acta Anatomica Sinica, 2018, 49(4):469-474. (in Chinese)

邓学军, 龚邵新, 杨胜辉, 等. MicroRNA-539靶向调控E2F转录因子3抑制肝癌的发生与发展［J］. 解剖学报, 2018, 49(4):469-474.

［4］Ren S, Xin Z, Xu Y, et al. Construction and analysis of circular RNA molecular regulatory networks in liver cancer［J］. Cell Cycle, 2017,16(22): 2204-2211.

［5］Zhang X, Xu X, Ge G, et al. miR-498 inhibits the growth and metastasis of liver cancer by targeting ZEB2［J］. Oncol Rep, 2019,41(3): 1638-1648.

［6］Zhou XJ, Wang XF, Feng YJ, et al. xpressions of SMG-1, ATM and P53 in laryngeal squamous cell carcinoma and their clinical significance［J］. Journal of Southern Medical University, 2016,36(1): 50-55. （in Chinese）

周学军, 王晓凤, 冯勇军, 等. 喉鳞癌组织中SMG-1、ATM和P53的表达及临床意义［J］. 南方医科大学学报, 2016,36(1): 50-55.

［7］Han LL, Nan HC, Tian T, et al. Expression and significance of the novel tumor-suppressor gene SMG-1 in hepatocellular carcinoma［J］. Oncol Rep, 2014,31(6): 2569-2578.

［8］Lin X, Chen Y. Identification of potentially functional circRNA-miRNA-mRNA regulatory network in hepatocellular carcinoma by integrated microarray analysis［J］. Med Sci Monit Basic Res, 2018,24(1): 70-78.

［9］Xiong DD, Dang YW, Lin P, et al. A circRNA-miRNA-mRNA network identification for exploring underlying pathogenesis and therapy strategy of hepatocellular carcinoma［J］. J Transl Med, 2018,16(1): 220-240.

［10］Yu J, Yang M, Zhou B, et al. CircRNA-104718 acts as competing endogenous RNA and promotes hepatocellular carcinoma progression through microRNA-218-5p/TXNDC5 signaling pathway［J］. Clin Sci, 2019,133(13): 1487-1503.

［11］Zhang PF, Wei CY, Huang XY, et al. Circular RNA circTRIM33-12 acts as the sponge of microRNA-191 to suppress hepatocellular carcinoma progression［J］. Mol Cancer, 2019,18(1): 105-119.

［12］Xing W, Zhou PC, Zhang HY, et al. Circular RNA circ_GLIS2 suppresses hepatocellular carcinoma growth and metastasis［J］. Liver Int, 2022, 42(3): 682-695.

［13］Wang H, Zhang Q, Cui W, et al. Circ_0004018 suppresses cell proliferation and migration in hepatocellular carcinoma via miR-1197/PTEN/PI3K/AKT signaling pathway［J］. Cell Cycle, 2021, 20(20):2125-2136.

［14］Li J, Bao S, Wang L, et al. CircZKSCAN1 suppresses hepatocellular carcinoma tumorigenesis by regulating miR-873-5p/downregulation of deleted in liver cancer 1［J］. Dig Dis Sci, 2021, 66(12):4374-4383.

［15］Zhou H, Chen Y. CircRNA has_circ_0001806 promotes hepatocellular carcinoma progression via the miR-193a-5p/MMP16 pathway［J］. Braz J Med Biol Res, 2021, 54(12):e11459.

［16］Chen Z, Jin P, Chen Z, et al. The expression of circ_0090049 in hepatocellular carcinoma and the molecular regulation mechanism of other biological functions［J］. Anticancer Drugs, 2022, 33(1): 48-60.

［17］Lu H, Gao L, Lv J. Circ_0078710 promotes the development of liver cancer by upregulating TXNDC5 via miR-431-5p［J］. Ann Hepatol, 2021, 27(1):100551.

［18］Liu C, Zhang W, Xing W, et al. MicroRNA-498 disturbs the occurrence and aggression of colon cancer through targeting MDM2 to mediate PPARγ ubiquitination［J］. Life Sci, 2021, 277:119225-119233.

［19］Gao N, Wang FX, Wang G, et al. Targeting the HMGA2 oncogene by miR-498 inhibits non-small cell lung cancer biological behaviors［J］. Eur Rev Med Pharmacol Sci, 2018,22(6): 1693-1699.

［20］Zhao T, Chen Y, Sheng S, et al. Upregulating microRNA-498 inhibits gastric cancer proliferation invasion and chemoresistance through inverse interaction of Bmi1［J］. Cancer Gene Ther, 2019,26(11-12): 366-373.

［21］Li W, Jiang H. Up-regulation of miR-498 inhibits cell proliferation, invasion and migration of hepatocellular carcinoma by targeting FOXO3［J］. Clin Res Hepatol Gastroenterol, 2020,44(1): 29-37.

［22］Yin DL, Lu PL, Yin RL, et al. Over-expression of PRRX1 induces apoptosis of hepatocellular carcinoma SMMC7721 cells through a p53-mediated mitochondrial apoptosis pathway［J］. Chinese Journal of Cancer Biotherapy, 2019,26(6): 662-668. （in Chinese）

尹东亮, 卢沛林, 尹润龙, 等. 过表达PRRX1 通过p53 介导的线粒体凋亡途径诱导肝癌SMMC7721 细胞凋亡［J］. 中国肿瘤生物治疗杂志, 2019,26(6): 662-668

［23］Gubanova E, Issaeva N, Gokturk C, et al. SMG-1 suppresses CDK2 and tumor growth by regulating both the p53 and Cdc25A signaling pathways［J］. Cell Cycle, 2013,12(24): 3770-3780.