Knockout of the HCC suppressor gene Lass2 downregulates the expression level of miR-694

  • Authors:
    • Xiaodong Lu
    • Yuanyuan Chen
    • Tiantian Zeng
    • Lufang  Chen
    • Qixiang Shao
    • Wenxin Qin
  • View Affiliations

  • Published online on: October 3, 2014
  • Pages: 2696-2702
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Homo sapiens longevity assurance homolog 2 of yeast LAG (Lass2) catalyzes the synthesis of long-chain ceramide which is an essential element of membranous structures. Deletion of Lass2 is associated with a high risk of spontaneous or DEN-induced hepatocellular carcinoma (HCC), yet the mechanism remains unclear. In the present study, we found extensive vesicles in hepatocytes of one-month-old Lass2-knockout (KO) mice. Hepatic biochemical indices were increased and expression of albumin was attenuated in the one‑month Lass2-KO liver. The results indicate that the injuries of the hepatocytes in young Lass2-KO mice, based on the results of Gene Ontology analysis of mRNA microarray of Lass2-KO liver vs. wild-type liver showed ‘wounding response’ was the mostly possible altered pathway in the Lass2-KO mice. miR-mRNA integrated analysis revealed that miR-694 was downregulated while its target gene tumor necrosis factor α-induced protein 3 (Tnfaip3) was upregulated, as confirmed by qPCR. The expression of NF-κB which is negatively controlled by Tnfaip3 was detected by qPCR and was found to be downregulated. Herein, we first report that Lass2 deficiency caused the downregulation of miR-694 and the upregulation of its target gene Tnfaip3 in vivo in mice, which may be related to a high risk of occurrence of HCC.


Recently Homo sapiens longevity assurance homolog 2 of yeast LAG (Lass2) has attracted the interest of researchers since a large amount of evidence has demonstrated that Lass2 is a potential tumor-suppressor gene. Deficiency of Lass2 is involved in the tumorigenesis of various types of tumors, especially hepatocellular carcinoma (HCC) (14). It has been reported that two of three non-specific Lass2-deleted mice presented liver cancer spontaneously when they were about nine months old (3). In our recently published study, using a hepatocellular-specific Lass2-knockout (KO) animal model, we found that Lass2-KO mice were more susceptible to the carcinogen DEN, i.e., DEN caused the Lass2-KO mice to develop liver tumors earlier and the tumors developed more rapidly (5).

To explore the biological functions of Lass2 and the related mechanisms it employs to suppress HCC, the structures and functions of the livers of the Lass2-KO mice and wild-type (WT) mice were compared. Meanwhile, microarrays of mRNAs and miRNAs of the livers from the two genotypes were performed and analyzed.

Materials and methods


The hepatocyte-specific Lass2-KO mice used in the present study were generated by the crossing of mice (C57BL/6J) carrying floxed the second exon of Lass2 and Albumin-Cre transgenic mice (C57BL/6J), as previously reported (5). All protocols for animal care and use were approved by the Regulations for the Administration of Affairs Concerning Experimental Animals (The State Science and Technology Commission of P.R. China, 1988), and the Animal Experimental Center of Jiangsu University was licensed for animal experiments. All of the mice were housed in pathogen-free (SPF) animal facilities under a standard 12-h-light/12-h-dark cycle. Animals received free access to water and commercial mouse chow throughout the present study. One-month-old male Lass2-KO and -WT mice were sacrificed by cervical dislocation. The mice for the experiments that followed were n=10/group. Liver weight/body weight were measured.

Biochemical estimation

The mice were sacrificed by cervical dislocation, and blood harvested from the inferior vena cava was centrifuged at 1,600 × g for 10 min at room temperature to obtain serum for hepatic biochemical estimation. The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) in serum were estimated using an AutoAnalyzer (Hitachi 7600, Japan) at the Affiliated Hospital of Jiangsu University.

Histological sections and staining

Liver tissues were immediately removed from the sacrificed mice, partly fixed in AAF (100% alcohol 85 ml, acetic acid 5 ml, formalin 10 ml) for morphological examination, and partly stored at −80°C for further use. The tissues were paraffin-embedded and sectioned (5-μm thick). Periodic acid-Schiff (PAS) staining was performed using a commercially available kit (cat. no. 0609A14; Shellfish Gamma Biotechnology Co., Ltd., Nanjing, China). Briefly, the sections were incubated in 0.5% periodic acid solution for 15 min, rinsed with distilled water, and exposed to Schiff’s reagent for 20 min followed by two 3-min exposures to 0.6% sodium metabisulfite.

Western blotting

Tissues were lysed in RIPA lysis buffer containing the protease inhibitor phenylmethanesulfonyl fluoride (cat. nos. P0013C and ST506; both from Beyotime Institute of Biotechnology). The cell extracts were centrifuged at 12,000 × g for 20 min at 4°C in a Beckman Avanti-30 centrifuge, and the supernatants were used for the experiments. The protein concentrations were determined with the BCA assay kit (cat. no. P0009; Beyotime Institute of Biotechnology). The equivalent tissue proteins (10 μg/lane) were subjected to electrophoresis on a Mini-Protean Tetra Electrophoresis System (165–8001; Bio-Rad, Hercules, CA, USA) and transferred onto PVDF membranes (Millipore, Bedford, MA, USA) via a semi-dry transfer system (Bio-Rad). The membranes were blocked with 5% non-fat milk for 1 h at room temperature in TBST [50 mM (pH 7.5) Tris, 0.9% NaCl and 0.1% Tween-20] and then incubated with a 1:200 dilution of rabbit anti-mouse/human albumin (cat. no. BS6520; Bioworld Technology, Inc., Dublin, OH, USA) overnight at 4°C. The membranes were washed and then incubated with peroxidase goat anti-rabbit antibody (cat. no. XR-9920; ProSci Inc., Poway, CA USA) for 1 h at room temperature and developed using the BeyoEcl Plus kit (cat. no. P0018; Beyotime Institute of Biotechnology) for 1 min, and then scanned by ChampChemi professional (SG2011; Beijing Sage Creation Science Co., Ltd., Beijing, China).

RNA isolation and qPCR analysis

Total RNA including miRNA from liver tissues was extracted by TRIzol (cat. no. 15596026; Invitrogen Corp., Carlsbad, CA, USA) or the miRNeasy Mini kit (cat. no. 217004; Qiagen, Hilden, Germany) according to the manufacturer’s suggestions. For mRNA qPCR, RNA was transcribed into cDNA using the QuantiTect reverse transcription kit and QuantiTect SYBR-Green PCR kits (cat. no. 205311 and no. 204243; both from Qiagen) according to the manufacturer’s protocols. For miRNA qPCR, RNA was transcribed into cDNA using the miScript Reverse II transcription kit (cat. no. 218161; Qiagen). The reaction component consisted of total RNA 1 μg, miScript HiSpec buffer 4 μl, Nucleics Mix 2 μl, miScript reverse transcriptase mix 2 μl, RNase-free H2O up to 20 μl. The reaction was carried out at 37°C for 60 min and 95°C for 5 min on the ABI PCR 9700 system (Applied Biosystems, Foster City, CA, USA). cDNA was diluted in 80 μl nuclease-free H2O for further application by LightCycler 480 SYBR-Green I master (Roche, Switzerland; cat. no. 04887352001). The reaction system for qPCR consisted of: LightCycler 480 SYBR-Green I Master 5 μl, forward primer 0.2 μl, reverse primer 0.2 μl, cDNA 1 μl, nuclease-free H2O 3.6 μl. PCR was run on ABI 7500 Fast (Applied Biosystems) and normalized against the expression of GAPDH or U6. The program was performed at 95°C for 10 min, 95°C for 10 sec plus 60°C for 30 sec for 40 cycles. For the melting curve evaluation, the temperature was slowly increased from 60°C to 97°C, and 5 acquisitions per °C were performed continuously. All samples were analyzed in triplicate. Relative expression was calculated using the comparative threshold cycle (Ct) method and was indicated as n-fold change = −(ΔCtKO − ΔCtWT). The primers for Tnfaip3, NF-κB and GAPDH were: for Tnfaip3, 5′-CAGCACCTAAG CCAACGAGT-3′ and 5′-TGGACCTGTCAATGTGTTCG-3′; for NF-κB, 5′-AGCTTATGCCGAACTTCTCG-3′ and 5′-GAC TCCGGGATGGAATGTAA-3′; for GAPDH, 5′-GCAAGG TCATCCCAGAG-3′ and 5′-AAGTCGCAGGAGACAAC-3′; for miR-694 and U6, the miRNA specific primers were respectively: 5′-CTGAAAATGTTGCCTGAAG-3′ and 5′-CAAGG ATGACACGCAAATTCG-3′, whereas the reverse primer was the manufacturer-provided miScript universal primer.

Microarrays of mRNA and miRNA

Liver tissues from 2 one-month male Lass2-KO mice and 2 one-month male WT mice were respectively subjected to the experiments of Mouse OneArray (MOA 2.0 ver.) and Mouse and Rat miRNA OneArray (MRmiOA 4.0 ver.) using Phalanx microarray platform by Oebiotech Co. (Shanghai, China). Briefly, RNA was extracted from liver tissues with TRIzol reagent (Invitrogen), whose quantity and purity were assessed using a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies Inc., Wilmington, DE, USA). The purity and integrity of each extracted RNA met the requirements: A260/A280 >1.6, A260/A230 >1 and RNA integrity number (RIN) value >5. Small RNA fraction indicated the abundance of RNA <200 nt compared with the overall RNA area from Agilent RNA 6000 Nano assay and was acceptable for the miRNA assay. Possibility of genomic DNA contamination was excluded by gel electrophoresis. Two micrograms of RNA from each group was respectively converted into cyanine-5 labeled target cRNA, hybridized to either Mouse OneArray or Mouse and Rat miRNA OneArray by the Affymetrix GeneChip fluidics station 450, and scanned with an Affymetrix GeneChip scanner 3000 7G. After normalization, differentially expressed mRNAs or miRNAs were established at log2 |Fold change| >1 and P<0.05.

For advanced data analysis, all biological replicates were pooled and calculated to identify differentially expressed genes based on the threshold of fold change and the P-value. The correlation of expression profiles between biological replicates and treatment conditions was demonstrated by unsupervised hierarchical clustering analysis. A subset of genes was selected for clustering analysis. An intensity filter was used to select genes where the difference between the maximum and minimum intensity values exceeded 4,000 among all microarrays. For this microarray project, the number of genes clustered was 272 genes. According to previously selected differentially expressed gene lists, Gene Ontology (GO) analysis was performed by Oebiotech Co. Targetscan 5.1 was utilized to predict miR-targeting mRNA. mRNA-miRNA integration analysis demonstrated potential mRNA targets with inverse expression alterations as their regulatory miRs displayed in the mRNA microarray or miRNA microarray.

Statistical analysis

Data were analyzed by the Student’s t-test. P<0.05 was considered to indicate a statistically significant difference, and P≤0.01 and P≤0.001 are indicated by relevant symbols in the figures and legends. qPCR and western blot analyses were repeated three times.


Structural alterations in the liver tissues from hepatocyte-specific Lass2-KO mice were noted in the PAS-stained sections

The average ratio of liver weight/body weight of the Lass2-KO mice was higher than the ratio in the control WT mice (Fig. 1A). Compared to the liver tissues from the WT mice, the hepatocytes of the Lass2-KO displayed abundant vesicles (Fig. 1B).

Liver functions and the expression of ALB are altered in Lass2-KO mice compared with the WT control

The production of ALB in the liver was attenuated in the Lass2-KO mice (Fig. 2A). The levels of ALT, AST and LDH in serum were respectively elevated in the Lass2-KO mice, indicating abnormal liver function in the Lass2-KO mice (Fig. 2B–D), in accordance with the structural injury of the Lass2-KO hepatocytes.

Profiles of mRNAs and miRNAs in the liver tissues are respectively reprogrammed in Lass2-KO mice vs. WT control

mRNA and miRNA microarray results showed that over 600 mRNA were markedly upregulated and over 700 genes were downregulated; whereas 13 miRNAs were markedly upregulated and 31 miRNAs were downregulated (Table I). According to the GO analysis, ‘response to wounding’ and ‘inflammatory response’ were among the top-10 altered pathways (Table II). miRNA-mRNA integrated analysis identified 4 upregulated and 3 downregulated miRNAs and their respective negatively controlled target genes (Table III).

Table I

Altered miRNAs in the Lass2-KO mouse liver tissues compared to the WT mouse liver tissues.

Table I

Altered miRNAs in the Lass2-KO mouse liver tissues compared to the WT mouse liver tissues.

Upregulated miRNAsDownregulated miRNAs
mmu-miR-1198-5pmmu-miR-1192 mmu-miR-467d-3p
mmu-miR-125b-5p mmu-miR-1224-5pmmu-miR-467f
mmu-miR-142-3p mmu-miR-1247-3pmmu-miR-483-5p
mmu-miR-199b-3p mmu-miR-1894-3pmmu-miR-5109
mmu-miR-199b-5p mmu-miR-1934-3p mmu-miR-669a-3p
mmu-miR-2137 mmu-miR-30c-1-3p mmu-miR-669o-3p
mmu-miR-2861mmu-miR-320-3p mmu-miR-669c-3p
mmu-miR-326-5pmmu-miR-346-3p mmu-miR-669p-3p
mmu-miR-34a-5p mmu-miR-466f-3p mmu-miR-694
mmu-miR-491-3p mmu-miR-466h-3pmmu-miR-711
mmu-miR-5130 mmu-miR-466i-3pmmu-miR-744-5p

Table II

Top 10 enrichment pathway terms from the Gene Ontology (GO) analysis (biological process).

Table II

Top 10 enrichment pathway terms from the Gene Ontology (GO) analysis (biological process).

Gene set nameGenes in overlapP-value
Response to wounding100.0407
Humoral immune response30.0588
Homeostasis of number of cells20.112
Viral genome replication20.122
Locomotory behavior50.122
Inflammatory response60.148
Development of primary sexual characteristics20.172
Response to external stimulus120.172
Jak-Stat cascade20.225
Viral infectious cycle20.235

Table III

miRNA-mRNA integrated analysis of the Lass2-KO mouse liver tissues.

Table III

miRNA-mRNA integrated analysis of the Lass2-KO mouse liver tissues.

A, Upregulated miRNAs and their downregulated predicted target genes

mmu-miR-2861mmu-miR-5130mmu-miR-142-3p mmu-miR-125b-5p

B, Downregulated miRNAs and their upregulated predicted target genes

mmu-miR-1192 mmu-miR-466f-3p mmu-miR-694

Serpinh1 (PAI-1)
qPCR confirms that miR-694 is markedly downregulated

The level of miR-694 was downregulated in the Lass2-KO liver tissue, as confirmed by qPCR (Fig. 3).

Tnfaip3 is markedly upregulated whereas NF-κB is downregulated

Tnfaip3, one of the markedly upregulated mRNAs in the Lass2-KO liver tissues, which is also a putative target gene of miR-694, was confirmed by qPCR (Fig. 4A). Its negatively controlled NF-κB was found to be downregulated (Fig. 4B).


Lass2 is a member of the Lass family, which is conserved among eukaryotes and is abundantly distributed in the liver, kidney and brain. The major function of Lass2 is synthesizing long-chain ceramide-C:24–26 (6). Ceramide serves as the precursor of a series of more complex sphingolipids. Short-chain ceramides function as a second messenger in a variety of cellular events, including apoptosis and differentiation (7,8), and regulate various cellular processes linked to cancer development, progression, metastasis and resistance to therapy (9,10). By comparison, long-chain ceramide, as the important element in constructing membranous structures, may regulate the cellular behavior via influencing the property of membranes. For example, ablation of Lass2 causes morphological alterations of the property of membranes (11,12). In the present study, numerous vesicles were noted in the hepatocytes from young Lass2-KO mice (Fig. 1), in accordance with previous reports from other researchers. Based on the attenuated production of albumin and increased hepatic biochemical indices in the Lass2-KO mice, (Fig. 2), it appears that the hepatocellular-specific Lass2-KO mice underwent hepatocellular injury even at an early age.

The microarrays of mRNAs and miRNAs of the Lass2-KO mice liver tissues vs. control demonstrated that miR-694 was upregulated, which was confirmed by qPCR. Moreover, the predicted target gene Tnfaip3 was found to be upregulated, as shown in the results of either the microarray of mRNA or qPCR. NF-κB, which is usually commonly negatively controlled by Tnfaip3 was found downregulated in the Lass2-KO mouse liver tissues. In our previous study (5), another target gene Serpinh1 (PAI-1) of miR-694 was found to be upregulated. The data strongly suggest that Lass2 deletion influences the expression level of miR-694.

However, the function of miR-694 is uncertain. According to miRNA-mRNA integrated analysis in this study, the down-regulated Lass2-related miR-694 elevated 25 mRNAs including Tnfaip3. Tnfaip3 is commonly considered as an inflammation suppressor (13) by inactivation of lymphocytes of suppressing NF-κB (14,15). Although it is commonly considered a tumor suppressor, overexpression of Tnfaip3 has also been reported in several non-lymphoma solid cancers, including HCC (1618). Our data suggest that Tnfaip3/NF-κB might play a role in inhibiting the inflammation and protecting injured hepatocytes caused by deletion of Lass2. In another report, Lass2-KO mice displayed resistance to LPS-induced liver injury (19), which might also be explained by the inhibitory immunity mediated by Tnfaip3/NF-κB.

The functions of other predicted target genes of miR-694 are listed in Table IV. According to published reports, miR-694 target genes are involved it the regulation of various important cellular events, including transcription, cell signal transduction, metabolism of lipid and glucose and trafficking, suggesting a diversity of regulatory functions of miR-694. miRNAs act as highly effective regulators of intracellular events. Moreover, via endocytosis or exocytosis of miRNA-containing vesicles, miRNAs modulate the extracellular milieu including stromal cells and extracellular matrix (20). For example, altered Tnfaip3/NF-κB may influence the immunocytes in the liver, or PAI-1 which was found to be upregulated, may regulate the synthesis of ECM in the liver (5), either of which is a predicted target gene of miR-694. The actual functions of miR-694 may likely be beyond what is listed in Table IV.

Table IV

Functions of the miR-694 predicted target genes.

Table IV

Functions of the miR-694 predicted target genes.

FunctionmiR-694 target gene (ref.)
TranscriptionMef2c (21)
Ncoa7 (22)
Tnfaip3 (13,14)
Elavl4 (23,24)
Cell cycleSka1 (25)
Nup153 (26)
Cell signal transductionGng2 (27)
Abi2 (28,29)
Pde5a (30)
Pde4d (31)
Metabolism regulatorTrib1 (3234)
Sgms1 (3537)
Srebf1 (38)
Vldlr (39)
TraffickingCD63 (40,41)
Man1c1 (42)
Nup153 (26)
Sgms1 (43)
ECM depositionFbln5 (44,45)
PAI-1 (46)
Immunity regulationTrib1 (47,48)
Tnfaip3 (13,14)

Overall, the present study first reports the attenuated expression level of miR-694 in Lass2-KO mouse liver tissue and its alteration of the Tnfaip3/NF-κB pathway. Our data strongly suggest that miR-694 functions in maintaining homeostasis of the liver and provide the basis to explore the functions of Lass2-related microRNAs.


This study was supported by the Grant from the State Key Laboratory of Oncogenes and Related Genes (no. 90-10-02, to X.L.) and Clinical Medicine Science & Technology Project of Jiangsu Province of China (no. BL2013024).



longevity assurance gene 2



miR or miRNA



tumor necrosis factor α-induced protein 3


nuclear transcription factor-κB


real-time quantitative PCR


hepatocellular carcinoma



Pewzner-Jung Y, Ben-Dor S and Futerman AH: When do Lasses (longevity assurance genes) become CerS (ceramide synthases)? Insights into the regulation of ceramide synthesis. J Biol Chem. 281:25001–25005. 2006. View Article : Google Scholar : PubMed/NCBI


Wang H, Wang J, Zuo Y, et al: Expression and prognostic significance of a new tumor metastasis suppressor gene LASS2 in human bladder carcinoma. Med Oncol. 29:1921–1927. 2012. View Article : Google Scholar : PubMed/NCBI


Fan S, Niu Y, Tan N, et al: LASS2 enhances chemosensitivity of breast cancer by counteracting acidic tumor microenvironment through inhibiting activity of V-ATPase proton pump. Oncogene. 32:1682–1690. 2013. View Article : Google Scholar : PubMed/NCBI


Imgrund S, Hartmann D, Farwanah H, et al: Adult ceramide synthase 2 (CERS2)-deficient mice exhibit myelin sheath defects, cerebellar degeneration, and hepatocarcinomas. J Biol Chem. 284:33549–33560. 2009. View Article : Google Scholar : PubMed/NCBI


Chen L, Lu X, Zeng T, et al: Enhancement of DEN-induced liver tumourigenesis in hepatocyte-specific Lass2-knockout mice coincident with upregulation of the TGF-β1-Smad4-PAI-1 axis. Oncol Rep. 31:885–893. 2014.PubMed/NCBI


Teufel A, Maass T, Galle PR, et al: The longevity assurance homologue of yeast lag1 (Lass) gene family (Review). Int J Mol Med. 23:135–140. 2009.PubMed/NCBI


Morad SAF and Cabot MC: Ceramide-orchestrated signalling in cancer cells. Nat Rev Cancer. 13:51–65. 2013. View Article : Google Scholar : PubMed/NCBI


Stiban J, Tidhar R and Futerman AH: Ceramide synthases: roles in cell physiology and signaling. Adv Exp Med Biol. 688:60–71. 2010. View Article : Google Scholar : PubMed/NCBI


Saddoughi SA and Ogretmen B: Diverse functions of ceramide in cancer cell death and proliferation. Adv Cancer Res. 117:37–58. 2010. View Article : Google Scholar : PubMed/NCBI


Liu J, Beckman BS and Foroozesh M: A review of ceramide analogs as potential anticancer agents. Future Med Chem. 5:1405–1421. 2013. View Article : Google Scholar : PubMed/NCBI


Park JW, Park WJ, Kuperman Y, Boura-Halfon S, Pewzner-Jung Y and Futerman AH: Ablation of very long acyl chain sphingolipids causes hepatic insulin resistance in mice due to altered detergent-resistant membranes. Hepatology. 57:525–532. 2013. View Article : Google Scholar : PubMed/NCBI


Silva LC, Ben David O, Pewzner-Jung Y, et al: Ablation of ceramide synthase 2 strongly affects biophysical properties of membranes. J Lipid Res. 53:430–436. 2012. View Article : Google Scholar : PubMed/NCBI


Ma A and Malynn BA: A20: linking a complex regulator of ubiquitylation to immunity and human disease. Nat Rev Immunol. 12:774–785. 2012. View Article : Google Scholar : PubMed/NCBI


Pujari R, Hunte R, Khan W and Shembade N: A20-mediated negative regulation of canonical NF-κB signaling pathway. Immunol Res. 57:166–171. 2013.PubMed/NCBI


Zhang F, Yang L and Li Y: The role of A20 in the pathogenesis of lymphocytic malignancy. Cancer Cell Int. 12:442012. View Article : Google Scholar : PubMed/NCBI


Wang M and Li S: Bladder polypoid cystitis-derived A20 associates with tumorigenesis. Cell Biochem Biophys. 67:669–673. 2013. View Article : Google Scholar : PubMed/NCBI


Hjelmeland AB, Wu Q, Wickman S, et al: Targeting A20 decreases glioma stem cell survival and tumor growth. PLoS Biol. 8:e10003192010. View Article : Google Scholar : PubMed/NCBI


Wang CM, Wang Y, Fan CG, et al: miR-29c targets TNFAIP3, inhibits cell proliferation and induces apoptosis in hepatitis B virus-related hepatocellular carcinoma. Biochem Biophys Res Commun. 411:586–592. 2011. View Article : Google Scholar : PubMed/NCBI


Ali M, Fritsch J, Zigdon H, Pewzner-Jung Y, Schütze S and Futerman AH: Altering the sphingolipid acyl chain composition prevents LPS/GLN-mediated hepatic failure in mice by disrupting TNFR1 internalization. Cell Death Dis. 4:e9292013. View Article : Google Scholar : PubMed/NCBI


Su Y, Li X, Ji W, et al: Small molecule with big role: MicroRNAs in cancer metastatic microenvironments. Cancer Lett. 344:147–156. 2014. View Article : Google Scholar : PubMed/NCBI


Cante-Barrett K, Pieters R and Meijerink JPP: Myocyte enhancer factor 2C in hematopoiesis and leukemia. Oncogene. 33:403–410. 2014. View Article : Google Scholar : PubMed/NCBI


Higginbotham KS, Breyer JP, Bradley KM, et al: A multistage association study identifies a breast cancer genetic locus at NCOA7. Cancer Res. 71:3881–3888. 2011. View Article : Google Scholar : PubMed/NCBI


Bronicki LM and Jasmin BJ: Emerging complexity of the HuD/ELAVl4 gene; implications for neuronal development, function, and dysfunction. RNA. 19:1019–1037. 2013. View Article : Google Scholar : PubMed/NCBI


Lee EK, Kim W, Tominaga K, et al: RNA-binding protein HuD controls insulin translation. Mol Cell. 45:826–835. 2012. View Article : Google Scholar : PubMed/NCBI


Ye AA and Maresca TJ: Cell division: kinetochores SKAdaddle. Curr Biol. 23:R122–R124. 2013. View Article : Google Scholar : PubMed/NCBI


Ullman KS, Shah S, Powers MA and Forbes DJ: The nucleoporin nup153 plays a critical role in multiple types of nuclear export. Mol Biol Cell. 10:649–664. 1999. View Article : Google Scholar : PubMed/NCBI


Matsuda T, Hashimoto Y, Ueda H, et al: Specific isoprenyl group linked to transducin gamma-subunit is a determinant of its unique signaling properties among G-proteins. Biochemistry. 37:9843–9850. 1998. View Article : Google Scholar


Grove M, Demyanenko G, Echarri A, et al: ABI2-deficient mice exhibit defective cell migration, aberrant dendritic spine morphogenesis, and deficits in learning and memory. Mol Cell Biol. 24:10905–10922. 2004. View Article : Google Scholar : PubMed/NCBI


Dai Z and Pendergast AM: Abi-2, a novel SH3-containing protein interacts with the c-Abl tyrosine kinase and modulates c-Abl transforming activity. Genes Dev. 9:2569–2582. 1995. View Article : Google Scholar : PubMed/NCBI


Kulkarni SK and Patil CS: Phosphodiesterase 5 enzyme and its inhibitors: update on pharmacological and therapeutical aspects. Methods Find Exp Clin Pharmacol. 26:789–799. 2004. View Article : Google Scholar : PubMed/NCBI


Lynch MJ, Baillie GS and Houslay MD: cAMP-specific phosphodiesterase-4D5 (PDE4D5) provides a paradigm for understanding the unique non-redundant roles that PDE4 isoforms play in shaping compartmentalized cAMP cell signalling. Biochem Soc Trans. 35:938–941. 2007. View Article : Google Scholar : PubMed/NCBI


Ishizuka Y, Nakayama K, Ogawa A, et al: TRIB1 downregulates hepatic lipogenesis and glycogenesis via multiple molecular interactions. J Mol Endocrinol. 52:145–158. 2014. View Article : Google Scholar : PubMed/NCBI


Angyal A and Kiss-Toth E: The tribbles gene family and lipoprotein metabolism. Curr Opin Lipidol. 23:122–126. 2012. View Article : Google Scholar : PubMed/NCBI


Dugast E, Kiss-Toth E, Soulillou JP, Brouard S and Ashton-Chess J: The Tribbles-1 protein in humans: roles and functions in health and disease. Curr Mol Med. 13:80–85. 2013. View Article : Google Scholar : PubMed/NCBI


Vacaru AM, Tafesse FG, Ternes P, et al: Sphingomyelin synthase-related protein SMSr controls ceramide homeostasis in the ER. J Cell Biol. 185:1013–1027. 2009. View Article : Google Scholar : PubMed/NCBI


Liu J, Zhang H, Li Z, et al: Sphingomyelin synthase 2 is one of the determinants for plasma and liver sphingomyelin levels in mice. Arterioscler Thromb Vasc Biol. 29:850–856. 2009. View Article : Google Scholar : PubMed/NCBI


Separovic D, Hanada K, Awad Maitah MIY, et al: Sphingomyelin synthase 1 suppresses ceramide production and apoptosis post-photodamage. Biochem Biophys Res Commun. 358:196–202. 2007. View Article : Google Scholar : PubMed/NCBI


Ruiz R, Jideonwo V, Ahn M, et al: Sterol regulatory element-binding protein-1 (SREBP-1) is required to regulate glycogen synthesis and gluconeogenic gene expression in mouse liver. J Biol Chem. 289:5510–5517. 2014. View Article : Google Scholar : PubMed/NCBI


Go GW and Mani A: Low-density lipoprotein receptor (LDLR) family orchestrates cholesterol homeostasis. Yale J Biol Med. 85:19–28. 2012.PubMed/NCBI


Pols MS and Klumperman J: Trafficking and function of the tetraspanin CD63. Exp Cell Res. 315:1584–1592. 2009. View Article : Google Scholar : PubMed/NCBI


Maecker HT, Todd SC and Levy S: The tetraspanin superfamily: molecular facilitators. FASEB J. 11:428–442. 1997.PubMed/NCBI


Scott DW and Patel RP: Endothelial heterogeneity and adhesion molecules N-glycosylation: implications in leukocyte trafficking in inflammation. Glycobiology. 23:622–633. 2013. View Article : Google Scholar : PubMed/NCBI


Subathra M, Qureshi A and Luberto C: Sphingomyelin synthases regulate protein trafficking and secretion. PLoS One. 6:e236442011. View Article : Google Scholar : PubMed/NCBI


Kapustin A, Stepanova V, Aniol N, et al: Fibulin-5 binds urokinase-type plasminogen activator and mediates urokinase-stimulated β1-integrin-dependent cell migration. Biochem J. 443:491–503. 2012.PubMed/NCBI


Choi J, Bergdahl A, Zheng Q, Starcher B, Yanagisawa H and Davis EC: Analysis of dermal elastic fibers in the absence of fibulin-5 reveals potential roles for fibulin-5 in elastic fiber assembly. Matrix Biol. 28:211–220. 2009. View Article : Google Scholar : PubMed/NCBI


Declerck PJ and Gils A: Three decades of research on plasminogen activator inhibitor-1: a multifaceted serpin. Semin Thromb Hemost. 39:356–364. 2013. View Article : Google Scholar : PubMed/NCBI


Satoh T, Kidoya H, Naito H, et al: Critical role of Trib1 in differentiation of tissue-resident M2-like macrophages. Nature. 495:524–528. 2013. View Article : Google Scholar : PubMed/NCBI


Dugast E, Kiss-Toth E, Docherty L, et al: Identification of Tribbles-1 as a novel binding partner of Foxp3 in regulatory T cells. J Biol Chem. 288:10051–10060. 2013. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

Volume 32 Issue 6

Print ISSN: 1021-335X
Online ISSN:1791-2431

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
Spandidos Publications style
Lu X, Chen Y, Zeng T, Chen L, Shao Q and Qin W: Knockout of the HCC suppressor gene Lass2 downregulates the expression level of miR-694. Oncol Rep 32: 2696-2702, 2014
Lu, X., Chen, Y., Zeng, T., Chen, L., Shao, Q., & Qin, W. (2014). Knockout of the HCC suppressor gene Lass2 downregulates the expression level of miR-694. Oncology Reports, 32, 2696-2702.
Lu, X., Chen, Y., Zeng, T., Chen, L., Shao, Q., Qin, W."Knockout of the HCC suppressor gene Lass2 downregulates the expression level of miR-694". Oncology Reports 32.6 (2014): 2696-2702.
Lu, X., Chen, Y., Zeng, T., Chen, L., Shao, Q., Qin, W."Knockout of the HCC suppressor gene Lass2 downregulates the expression level of miR-694". Oncology Reports 32, no. 6 (2014): 2696-2702.