Expression of long-chain non-coding RNA FOXN3-AS2 in hepatocellular carcinoma and its effect on proliferation and invasion of hepatoma cells

doi:10.12092/j.issn.1009-2501.2018.11.008

Abstract

Abstract:

AIM: To observe the expression levels of FOXN3-AS2 in hepatocellular tissues and cells, and its effect on proliferation and invasion of hepatoma cells. METHODS: The expression levels of FOXN3-AS2 in 12 hepatocellular carcinoma tissues and 5 hepatocellular carcinoma cell lines were detected by real-time quantitative PCR (qRT-PCR). Transfection of plasmid in the hepatocellular carcinoma cell line with the lowest expression level of FOXN3-AS2 was used to increase the expression of FOXN3-AS2. The cell counting kit (CCK-8) and Transwell invasion assay were used to detect the effect of overexpressed FOXN3-AS2 on proliferation and invasion of hepatoma cells. Bioinformatics was used to predict the miRNA that FOXN3-AS2 could complement and related genes, qRT-PCR detected the expression levels of miRNA and related gene mRNA, and Western blot was used to detect the expression level of related proteins. RESULTS: The expression level of FOXN3-AS2 in hepatocellular carcinoma tissues was significantly lower than that in adjacent tissues (P<0.01). The expression level of FOXN3-AS2 in hepatocellular carcinoma cell lines was significantly lower than that in human normal liver cells (P<0.05). FOXN3-AS2 has the lowest expression level in HepG2 cells (P<0.01). High expression of FOXN3-AS2 significantly inhibited the proliferation activity (P<0.05) and invasive ability (P<0.05). FOXN3-AS2 could complement the miR-34a-5p, and miR-34a-5p can complement the KLF4 gene. FOXN3-AS2 can complementarily pair with miR-34a-5p, and miR-34a-5p can complementarily pair with KLF4. High expression of FOXN3-AS2 decreased the expression of miR-34a-5p (P<0.01) and increased the expression of KLF4 mRNA (P<0.01). The expression of KLF4, β-catenin and E-cadherin proteins were increased, while the expression of CDK4 and Cyclin D1 proteins were decreased. CONCLUSION: The expression of FOXN3-AS2 is down-regulated in HCC tissues and cells. The high expression of FOXN3-AS2 can inhibit the proliferation and invasion of HepG2 cells. The mechanism may be related with regulating the expression of miR-34a-5p and KLF4 genes.

Key words: hepatocellular carcinoma, long non-coding RNA, cell proliferation, cell invasion

CLC Number:

CAI Min, XU Liu, SHEN Lan, ZHANG Jie. Expression of long-chain non-coding RNA FOXN3-AS2 in hepatocellular carcinoma and its effect on proliferation and invasion of hepatoma cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(11): 1246-1251.

References

［1］Zhang Y, Wen DY, Zhang R, et al. A Preliminary investigation of PVT1 on the effect and mechanisms of hepatocellular carcinoma: evidence from clinical data, a Meta-Analysis of 840 cases, and in vivo validation［J］. Cell Physiol Biochem,2018,47(6):2216-2232.
［2］Ding H, Liu J, Liu B, et al. Long noncoding RNA PVT1 inhibits interferon-alpha mediated therapy for hepatocellular carcinoma cells by interacting with signal transducer and activator of transcription 1［J］. Biochem Biophys Res Commun,2018,500(4):973-980.
［3］Su W, Feng S, Chen X, et al. Silencing of long noncoding RNA MIR22HG triggers cell survival/death signaling via oncogenes YBX1, MET, and p21 in lung cancer［J］. Cancer Res,2018,78(12):3207-3219.
［4］Yu WW, Wang K, Liao GJ. Knockdown of long noncoding RNA linc-ITGB1 suppresses migration, invasion of hepatocellular carcinoma via regulating ZEB1［J］. Eur Rev Med Pharmacol Sci,2017,21(22):5089-5095.
［5］张蕊，邓俊丽，孙红，等. 长链非编码RNA对肿瘤转移调控的研究进展［J］. 中国临床药理学与治疗学,2016,21(9):1061-1067.
［6］Li T, Liu Y, Sun Y. Long non-coding RNA AB209630 suppresses cell proliferation and metastasis in human hepatocellular carcinoma［J］. Exp Ther Med,2017,14(4):3419-3424.
［7］孙跃胜，屈强，潘江华，等. 长链非编码RNA在结直肠癌中的作用［J］. 中国临床药理学与治疗学,2017,22(2):222-227.
［8］Huang J, Deng G, Liu T, et al. Long noncoding RNA PCAT-1 acts as an oncogene in osteosarcoma by reducing p21 levels［J］. Biochem Biophys Res Commun,2018,495(4):2622-2629.
［9］Cao SW, Huang JL, Chen J, et al. Long non-coding RNA UBE2CP3 promotes tumor metastasis by inducing epithelial-mesenchymal transition in hepatocellular carcinoma［J］. Oncotarget,2017,8(39):65370-65385.
［10］Zhang Y, Huang JC, Cai KT, et al. Long noncoding RNA HOTTIP promotes hepatocellular carcinoma tumorigenesis and development: A comprehensive investigation based on bioinformatics, qRTPCR and metaanalysis of 393 cases［J］. Int J Oncol,2017,51(6):1705-1721.
［11］Wang J, Ma W, Liu Y. Long non-coding RNA HULC promotes bladder cancer cells proliferation but inhibits apoptosis via regulation of ZIC2 and PI3K/AKT signaling pathway［J］. Cancer Biomark,2017,20(4):425-434.
［12］Hu B, Cai H, Zheng R, et al. Long non-coding RNA 657 suppresses hepatocellular carcinoma cell growth by acting as a molecular sponge of miR-106a-5p to regulate PTEN expression［J］. Int J Biochem Cell Biol,2017,92:34-42.
［13］Lin X, Lin BW, Chen XL, et al. PAI-1/PIAS3/Stat3/miR-34a forms a positive feedback loop to promote EMT-mediated metastasis through Stat3 signaling in Non-small cell lung cancer［J］. Biochem Biophys Res Commun,2017,493(4):1464-1470.
［14］Pu Y, Zhao F, Wang H, et al. MiR-34a-5p promotes the multi-drug resistance of osteosarcoma by targeting the CD117 gene［J］. Oncotarget,2016,7(19):28420-28434.
［15］Ding X, Zhong T, Jiang L, et al. miR-25 enhances cell migration and invasion in non-small-cell lung cancer cells via ERK signaling pathway by inhibiting KLF4［J］. Mol Med Rep,2018,17(5):7005-7016.
［16］Kong F, Sun T, Kong X, et al. Kruppel-like factor 4 suppresses serine/threonine kinase 33 activation and metastasis of gastric cancer through reversing epithelial-mesenchymal transition［J］. Clin Cancer Res,2018,24(10):2440-2451.
［17］Xu X, Li J, Zhu Y, et al. CRISPR-ON-Mediated KLF4 overexpression inhibits the proliferation, migration and invasion of urothelial bladder cancer in vitro and in vivo［J］. Oncotarget,2017,8(60):102078-102087.
［18］Sun H, Peng Z, Tang H, et al. Loss of KLF4 and consequential downregulation of Smad7 exacerbate oncogenic TGF-beta signaling in and promote progression of hepatocellular carcinoma［J］. Oncogene,2017,36(21):2957-2968.

[1]	WU Xianchuang, LIU Yuxin, NIU Yuji, HE Jinjin, QIAO Hui, ZHANG Leilei. Mechanism of DCLK1 transcriptional regulation in HCC [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(11): 1241-1246.
[2]	DING Jiamin, WANG Youdi, ZHAO Xiali, JIANG Genfeng, CHEN Siwen, HONG Chengcheng, LI Shuqin, HU Weihua. Study on the treatment of thin endometrium with growth hormone and vitamin E [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2022, 27(8): 857-862.
[3]	WANG Xinli, XU Xiaqing, FANG Hanbing, GUO Yuzhong. Study of TPA on enhancing the anti-tumor effects of cisplatin and reducing its renal toxicity [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2022, 27(5): 535-543.
[4]	LI Yanyan, FANG Xiaojie, YIN Xin, SUN Xi, RAO Chunhui. circZNF124 regulates the proliferation, migration and invasion of colorectal cancer SW620 cells by targeting miR-4262 [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2022, 27(11): 1231-1239.
[5]	YE Kaili, ZHENG Wen, YE Qifa, YANG Lan . Mechanism of CDK1 participates in the development of hepatocellular carcinoma and its inhibitor application value [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2021, 26(9): 1086-1094.
[6]	YU Saihong, ZHENG Xiaoliang, PU Yiyi, YAN Dongmei, WANG Xiaoju, YU Jie. Reversal of 5-fluorouracil resistance in hepatocellular carcinoma cells by inhibiting ribonucleotide reductase M2 [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2021, 26(7): 729-737.
[7]	SONG Ruzheng, PENG Ying, WANG Guangji, SUN Jianguo. Commonly used quantitative proteomics research techniques and their application in the study of pathogenesis of liver-derived diseases [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2021, 26(5): 570-578.
[8]	JIN Le, JIANG Duochen, CHEN Hongxiao, LIU Su, CHEN Zhaolin, JU Jing. Quercetin reverses adriamycin resistance by activating GSK-3β/β-catenin pathways through GAS5 in breast cancer cells [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2021, 26(12): 1344-1351.
[9]	PENG Fusheng, HUANG Xiaohui, LI Peng, TANG Jian'er. miR-34a inhibits proliferation of prostate cancer LNCaP cells by regulating androgen receptor gene [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2021, 26(1): 10-17.
[10]	SONG Dandan, CHEN Xueyuan, CHEN Ruiqi, LI Hanqiao, WU Tian. Research status of long non-coding RNA MALAT1 in breast cancer [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2021, 26(1): 49-57.
[11]	WANG Xianghai, XING Min, LYU Tangfeng, LIU Hongbing, SONG Yong. Effects of knocking down long intergenic non-coding RNA 00467 on the prognosis of patients with lung adenocarcinoma and its mechanism [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2020, 25(8): 841-849.
[12]	HOU Bin, HUANG Weiping, YIN Zhongpu, HU Shousen. lncRNA PCAT19 inhibits the proliferation and invasion of nasopharyngeal carcinoma cells by adsorbing miR-142-5p and regulating the expression of ING3 gene [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2020, 25(7): 721-727.
[13]	WANG Ziyuan, SUN Mingyu. Advances in research on long non-coding RNA in drug resistance of colorectal cancer [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2020, 25(7): 796-802.
[14]	HUANG Kang, LI Ling, YE Qifa, PENG Guizhu. Expression of long non-coding RNA LINC00844 in hepatocellular carcinoma and its inhibiting effect on cell proliferation and migration [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2020, 25(4): 366-372.
[15]	CHENG Jianping, LI Zhen, ZHAO Xiaolin, YANG Mengyuan, JI Xiwei, YU Jiufei. Research on the expression of miR-186-5p in serum of patients with colon cancer#br# [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2020, 25(10): 1097-1104.