Oleate acid-stimulated HMMR expression by CEBPα is associated with nonalcoholic steatohepatitis and hepatocellular carcinoma

Abstract

Non-alcoholic steatohepatitis (NASH) is a type of nonalcoholic fatty liver disease and has become a major risk factor for hepatocellular carcinoma (HCC). However, the underlying pathophysiological mechanisms are still elusive. Here, we identify hyaluronan-mediated motility receptor (HMMR) as a critical gene associated with NASH/HCC by combination of bioinformatic analysis and functional experiments. Analysis of differentially expressed genes (DEGs) between normal controls and NASH/HCC identified 5 hub genes (HMMR, UBE2T, TYMS, PTTG1 and GINS2). Based on the common DEGs, analyses of univariate and multivariate Cox regression and the area under the curve (AUC) value of the receiver operating characteristic (ROC) indicate that HMMR is the most significant gene associated with NASH/HCC among five hub genes. Oleate acid (OA), one of fatty acids that induce cellular adipogenesis, stimulates HMMR expression via CCAAT/enhancer-binding protein α (CEBPα). CEBPα increases the expression of HMMR through binding to its promoter. HMMR promotes HCC cell proliferation in vitro via activation of G1/S and G2/M checkpoint transitions, concomitant with a marked increase of the positive cell cycle regulators, including cyclin D1, cyclin E, and cyclin B1. Knockdown of HMMR suppresses HCC tumor growth in nude mice. Our study identifies an important role of HMMR in NASH/HCC, and suggests that HMMR may be a useful target for therapy and prognostic prediction of NASH/HCC patients.

RNA Sequencing Analysis of Gene Expression by Electroacupuncture in Guinea Pig Gallstone Models

“…By analyzing microarray data, UBE2T was considered a hub gene in patients with gallbladder cancer, and it might serve as a biomarker for gallbladder cancer [50]. rough the PI3K/Akt signaling pathway and the Akt/GSK3β/β-catenin pathway, UBE2T is involved in the cell proliferation, migration, and invasion of cancer cells [51][52][53]. UBE2NL, also a ubiquitin-conjugating enzyme E2 family member, is related to the ubiquitination process [54].…”

Knockdown of FBI-1 Inhibits the Warburg Effect and Enhances the Sensitivity of Hepatocellular Carcinoma Cells to Molecular Targeted Agents via miR-3692/HIF-1α

“…The RNA samples from the cells or the tumor tissues were extracted and reverse transcribed into cDNA according to the manufacturer’s instructions (Thermo Fisher Scientific, Waltham, MA, USA) ( 26 – 28 ). Then, the cDNA samples were analyzed by quantitative PCR (qPCR) according to the manufacturer’s instructions (Thermo Fisher Scientific, Waltham, MA, USA) and methods described in previous publications ( 26 – 28 ). Primers used in the present work were listed as follows: (1) FBI-1, forward sequence, 5’-GCAACATCTGCAAGGTCCGCTT-3’; reverse sequence, 5’-TCTTCAGGTCGTAGTTGTGGGC-3’; (2) miR-3692-5p, the reverse-transcription primer, 5’-GTCGTATCCAGTGCGTGTCGTGGAGTCGGCAATTGCACTGGATACGACCAGTAT-3’; forward sequence, 5’-CCTGCTGGTCAGGAGTGGATACTG-3’; reverse sequence, 5’-CAGTGCGTGTCGTGGAGT-3’; (3) HIF-1α, forward sequence, 5’-TATGAGCCAGAAGAACTTTTAGGC-3’; reverse sequence, 5’-CACCTCTTTTGGCAAGCATCCTG-3’; (4) GLUT1, forward sequence, 5’-TTGCAGGCTTCTCCAACTGGAC-3’; reverse sequence, 5’-CAGAACCAGGAGCACAGTGAAG-3’; (5) LDHA, forward sequence, 5’-GGATCTCCAACATGGCAGCCTT-3’; reverse sequence, 5’-AGACGGCTTTCTCCCTCTTGCT-3’; (6) N-cadherin, forward sequence, 5’-CCTCCAGAGTTTACTGCCATGAC-3’; reverse sequence, 5’-GTAGGATCTCCGCCACTGATTC-3’; (7) E-cadherin, forward sequence, 5’-GCCTCCTGAAAAGAGAGTGGAAG-3’; reverse sequence, 5’-TGGCAGTGTCTCTCCAAATCCG-3’; and (7) Vimentin, forward sequence, 5’-AGGCAAAGCAGGAGTCCACTGA-3’; reverse sequence, 5’-ATCTGGCGTTCCAGGGACTCAT-3’.…”

“…For the animal experiments, the subcutaneous tumor tissues were collected and grinded by using the liquid nitrogen. The RNA samples from the cells or the tumor tissues were extracted and reverse transcribed into cDNA according to the manufacturer's instructions (Thermo Fisher Scientific, Waltham, MA, USA) (26)(27)(28). Then, the cDNA samples were analyzed by quantitative PCR (qPCR) according to the manufacturer's instructions (Thermo Fisher Scientific, Waltham, MA, USA) and methods described in previous publications (26)(27)(28).…”

“…The RNA samples from the cells or the tumor tissues were extracted and reverse transcribed into cDNA according to the manufacturer's instructions (Thermo Fisher Scientific, Waltham, MA, USA) (26)(27)(28). Then, the cDNA samples were analyzed by quantitative PCR (qPCR) according to the manufacturer's instructions (Thermo Fisher Scientific, Waltham, MA, USA) and methods described in previous publications (26)(27)(28). Primers used in the present work were listed as follows: (1) FBI-1, forward sequence, 5'-GCAACA T C T G C A A G G T C C G CT T -3 ' ; r e v e r s e s e q u e n c e , 5' -TCTTCAGGTCGTAGTTGTGGGC-3'; (2) miR-3692-5p, the reverse-transcription primer, 5 '…”

Table6.DOCX

“…However, very little research can be found about GINS2 in the liver, especially in NAFLD. Previous bioinformatics studies indicated that GINS2 might be the hub genes in the development of NASH to HCC and predicted poor prognosis in HCC, but there was no further experiment to verify its effects on NAFLD (Lian et al, 2018;Zhang et al, 2020). PDLIM3, highly expressed in skeletal and cardiac muscle, has been suggested to play a pivotal role in myocyte stability, signal transduction, and mechanical signaling, especially in growth and remodeling processes (Zheng et al, 2010).…”