Effect of long non-coding RNA LNC 01309 on proliferation and migration abilities of human hepatoma cells and its mechanism of action

Hongyan LIU; Jingjiao LI; Zejun LU; Yongzhen LIU; Juyi WEN

doi:10.3969/j.issn.1001-5256.2022.03.014

Volume 38 Issue 3

Mar. 2022

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Article Navigation > Journal of Clinical Hepatology > 2022 > 38(3): 563-571

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Effect of long non-coding RNA LNC 01309 on proliferation and migration abilities of human hepatoma cells and its mechanism of action

DOI: 10.3969/j.issn.1001-5256.2022.03.014

1.
Naval Clinical College of Anhui Medical University, Hefei 230032, China
2.
Department of Oncology, The Sixth Medical Center of PLA General Hospital, Beijing 100048, China

Research funding:

The Key Logistics Science Research Project of PLA (BHJ14C008)

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Corresponding author: WEN Juyi, wenjuyi@126.com(ORCID:0000-0002-7469-7811)
Received Date: 2021-07-21
Accepted Date: 2021-09-29
Published Date: 2022-03-20

Abstract

Abstract

Objective To investigate the effect of long non-coding RNA (lncRNA) LNC01309 on the proliferation and migration abilities of human hepatocellular carcinoma (HCC) cells and its mechanism of action. Methods HCC samples and corresponding adjacent tissue samples were collected from 12 patients with HCC who underwent surgical treatment in The Sixth Medical Center of PLA General Hospital from February 2018 to June 2019, and quantitative real-time PCR was used to measure the relative expression level of LNC01309. Quantitative real-time PCR was also used to measure the expression level of LNC01309 in human hepatoma cell lines (HepG2, SNU-398, and Hep3B) and the human immortalized normal liver cell line THLE-2. After LNC01309 was overexpressed in HepG2 cells, the cells were divided into plasmid control group (pEXP-control) and overexpression group (pEXP-LNC01309). CCK-8 assay was used to observe the change in cell proliferation, and wound healing assay and Transwell assay were used to observe migration ability. RNA co-immunoprecipitation was used to detect the interaction between LNC01309 with RBM38, with cells divided into IgG group and RBM38 antibody group, and cycloheximide chase assay was used to analyze the effect of LNC01309 on the stability of RBM38 protein. RBM38 was overexpressed in HepG2 cells to conduct the recovery experiment, and CCK-8 assay, wound healing assay, and Transwell assay were used to observe the changes in cell proliferation and migration abilities. The t-test was used for comparison of continuous data between two groups. Results The mean expression level of LNC01309 in HCC tissue was significantly higher than that in adjacent tissue (4.225±2.285 vs 1.541±0.530, t=3.618, P=0.004), and the relative expression level of LNC01309 in hepatoma cells (HepG2, SNU-398, and Hep3B) was also significantly higher than that in normal hepatocytes (THLE-2) (t=4.231、6.489、14.480, all P < 0.05). Compared with the control group, HepG2 cells with the overexpression of LNC01309 had significant increases in growth rate (OD₄₅₀ value at 96 hours: 1.885±0.107 vs 2.527±0.234, t=4.330, P=0.012) and migration ability (11.65%±2.40% vs 35.66%±4.90%, t=9.837, P < 0.001; 100.00%±3.11% vs 161.00%±35.93%, t=4.399, P=0.005); however, the upregulated proliferation and migration abilities of hepatoma cells induced by LNC01309 overexpression were partially inhibited by RBM38 (OD₄₅₀ value at 96 hours: 2.500±0.227 vs 1.913±0.282, t=2.812, P=0.048; 168.00%±9.43% vs 117.20%±18.03%, t=6.622, P < 0.001). Compared with the IgG control group, RBM38 antibody significantly enriched the precipitation of LNC01309 (t=3.846, P=0.031). The results of cycloheximide chase assay showed that the LNC01309 overexpression group had a significant reduction in the stability of RBM38 protein (t=8.038, P=0.001). Conclusion The newly identified LNC01309 reduces the stability of RBM38 protein through interaction with RBM38 and promotes the proliferation and migration of HCC cells.
- Carcinoma, Hepatocellular,
- RNA, Long Noncoding,
- RNA-Binding Proteins

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References

[1]	BHAN A, SOLEIMANI M, MANDAL SS. Long noncoding RNA and cancer: A new paradigm[J]. Cancer Res, 2017, 77(15): 3965-3981. DOI: 10.1158/0008-5472.CAN-16-2634.
[2]	PENG WX, KOIRALA P, MO YY. LncRNA-mediated regulation of cell signaling in cancer[J]. Oncogene, 2017, 36(41): 5661-5667. DOI: 10.1038/onc.2017.184.
[3]	MATTICK JS. The genetic signatures of noncoding RNAs[J]. PLoS Genet, 2009, 5(4): e1000459. DOI: 10.1371/journal.pgen.1000459.
[4]	KUMAR S, GONZALEZ EA, RAMESHWAR P, et al. Non-coding RNAs as mediators of epigenetic changes in malignancies[J]. Cancers (Basel), 2020, 12(12): 3657. DOI: 10.3390/cancers12123657.
[5]	MA L, BAJIC VB, ZHANG Z. On the classification of long non-coding RNAs[J]. RNA Biol, 2013, 10(6): 925-933. DOI: 10.4161/rna.24604.
[6]	MIYAMOTO M, MOTOOKA D, GOTOH K, et al. Performance comparison of second- and third-generation sequencers using a bacterial genome with two chromosomes[J]. BMC Genomics, 2014, 15: 699. DOI: 10.1186/1471-2164-15-699.
[7]	RHOADS A, AU KF. PacBio sequencing and its applications[J]. Genomics Proteomics Bioinformatics, 2015, 13(5): 278-289. DOI: 10.1016/j.gpb.2015.08.002.
[8]	XU J, ZHENG Y, PU S, et al. Third-generation sequencing found LncRNA associated with heat shock protein response to heat stress in Populus qiongdaoensis seedlings[J]. BMC Genomics, 2020, 21(1): 572. DOI: 10.1186/s12864-020-06979-z.
[9]	SHEN Y, LIANG W, LIN Y, et al. Single molecule real-time sequencing and RNA-seq unravel the role of long non-coding and circular RNA in the regulatory network during Nile tilapia (Oreochromis niloticus) infection with Streptococcus agalactiae[J]. Fish Shellfish Immunol, 2020, 104: 640-653. DOI: 10.1016/j.fsi.2020.06.015.
[10]	HUANG Z, ZHOU JK, PENG Y, et al. The role of long noncoding RNAs in hepatocellular carcinoma[J]. Mol Cancer, 2020, 19(1): 77. DOI: 10.1186/s12943-020-01188-4.
[11]	CHEN Y, LI Z, CHEN X, et al. Long non-coding RNAs: From disease code to drug role[J]. Acta Pharm Sin B, 2021, 11(2): 340-354. DOI: 10.1016/j.apsb.2020.10.001.
[12]	LI H, JI FX, XU XN, et al. Effects of long-chain non-coding RNA LINC00467 targeting miR-1247-5p on proliferation, migration and invasion of hepatocellular carcinoma cells[J]. Chin J Gerontol, 2021, 41(15): 3328-3333. DOI: 10.3969/j.issn.1005-9202.2021.15.048 李豪, 姬发祥, 徐晓宁, 等. 长链非编码RNA LINC00467靶向miR-1247-5p调控肝癌细胞增殖、迁移及侵袭的影响[J]. 中国老年学杂志, 2021, 41(15): 3328-3333. DOI: 10.3969/j.issn.1005-9202.2021.15.048
[13]	YUAN JH, YANG F, WANG F, et al. A long noncoding RNA activated by TGF-β promotes the invasion-metastasis cascade in hepatocellular carcinoma[J]. Cancer Cell, 2014, 25(5): 666-681. DOI: 10.1016/j.ccr.2014.03.010.
[14]	PANZITT K, TSCHERNATSCH MM, GUELLY C, et al. Characterization of HULC, a novel gene with striking up-regulation in hepatocellular carcinoma, as noncoding RNA[J]. Gastroenterology, 2007, 132(1): 330-342. DOI: 10.1053/j.gastro.2006.08.026.
[15]	DU Y, KONG G, YOU X, et al. Elevation of highly up-regulated in liver cancer (HULC) by hepatitis B virus X protein promotes hepatoma cell proliferation via down-regulating p18[J]. J Biol Chem, 2012, 287(31): 26302-26311. DOI: 10.1074/jbc.M112.342113.
[16]	GENG YJ, XIE SL, LI Q, et al. Large intervening non-coding RNA HOTAIR is associated with hepatocellular carcinoma progression[J]. J Int Med Res, 2011, 39(6): 2119-2128. DOI: 10.1177/147323001103900608.
[17]	LI G, ZHANG H, WAN X, et al. Long noncoding RNA plays a key role in metastasis and prognosis of hepatocellular carcinoma[J]. Biomed Res Int, 2014, 2014: 780521. DOI: 10.1155/2014/780521.
[18]	MAASS PG, LUFT FC, BÄHRING S. Long non-coding RNA in health and disease[J]. J Mol Med (Berl), 2014, 92(4): 337-346. DOI: 10.1007/s00109-014-1131-8.
[19]	MENG YM, JIANG J, WANG YH, et al. Mechanism of long noncoding RNA encoded peptide in tumor[J]. Clin Misdiagn Misther, 2021, 34(9): 113-116. https://www.cnki.com.cn/Article/CJFDTOTAL-LCWZ202109024.htm 孟一妹, 蒋晶, 王禹涵, 等. 长链非编码RNA编码肽在肿瘤中的作用机制[J]. 临床误诊误治, 2021, 34(9): 113-116. https://www.cnki.com.cn/Article/CJFDTOTAL-LCWZ202109024.htm
[20]	SCHAUKOWITCH K, KIM TK. Emerging epigenetic mechanisms of long non-coding RNAs[J]. Neuroscience, 2014, 264: 25-38. DOI: 10.1016/j.neuroscience.2013.12.009.
[21]	HUANG JZ, CHEN M, CHEN, et al. A peptide encoded by a putative lncRNA HOXB-AS3 suppresses colon cancer growth[J]. Mol Cell, 2017, 68(1): 171-184. DOI: 10.1016/j.molcel.2017.09.015.
[22]	LI XX, SHI L, ZHOU XJ, et al. The role of c-Myc-RBM38 loop in the growth suppression in breast cancer[J]. J Exp Clin Cancer Res, 2017, 36(1): 49. DOI: 10.1186/s13046-017-0521-5.
[23]	JIANG Y, XU E, ZHANG J, et al. The Rbm38-p63 feedback loop is critical for tumor suppression and longevity[J]. Oncogene, 2018, 37(21): 2863-2872. DOI: 10.1038/s41388-018-0176-5.
[24]	CHO SJ, ZHANG J, CHEN X. RNPC1 modulates the RNA-binding activity of, and cooperates with, HuR to regulate p21 mRNA stability[J]. Nucleic Acids Res, 2010, 38(7): 2256-2267. DOI: 10.1093/nar/gkp1229.
[25]	XU E, ZHANG J, CHEN X. MDM2 expression is repressed by the RNA-binding protein RNPC1 via mRNA stability[J]. Oncogene, 2013, 32(17): 2169-2178. DOI: 10.1038/onc.2012.238.
[26]	YAN W, ZHANG J, ZHANG Y, et al. p73 expression is regulated by RNPC1, a target of the p53 family, via mRNA stability[J]. Mol Cell Biol, 2012, 32(13): 2336-2348. DOI: 10.1128/MCB.00215-12.
[27]	SUN Z, YANG GS, SIMA H, et al. Expression of RBM5 and its effect on prognosis of hepatocellular carcinoma patients after hepatectomy[J/CD]. Chin J Hepat Surg (Electronic Edition), 2021, 10(1): 93-97. DOI: 10.3877/cma.j.issn.2095-3232.2021.01.020. 孙喆, 杨广顺, 司马辉, 等. RBM5在肝细胞癌中的表达及其对肝切除术后患者预后的影响[J/CD]. 中华肝脏外科手术学电子杂志, 2021, 10(1): 93-97. DOI: 10.3877/cma.j.issn.2095-3232.2021.01.020.
[28]	YE J, LIANG R, BAI T, et al. RBM38 plays a tumor-suppressor role via stabilizing the p53-mdm2 loop function in hepatocellular carcinoma[J]. J Exp Clin Cancer Res, 2018, 37(1): 212. DOI: 10.1186/s13046-018-0852-x.

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Effect of long non-coding RNA LNC 01309 on proliferation and migration abilities of human hepatoma cells and its mechanism of action

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Effect of long non-coding RNA LNC 01309 on proliferation and migration abilities of human hepatoma cells and its mechanism of action

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