[Retracted] miR­365 induces hepatocellular carcinoma cell apoptosis through targeting Bcl­2.

[This retracts the article DOI: 10.3892/etm.2017.4244.].

Thioredoxin-interacting protein: a critical link between autophagy disorders and pancreatic ß-cell dysfunction.

Thioredoxin-interacting protein (TXNIP) is a known important regulatory protein of islet ß-cell biology and function, but the detailed mechanism is not clear. Autophagy plays a pivotal role in maintaining cellular homoeostasis. This study aimed to elucidate the influence of TXNIP on the autophagy of ß-cell. In this study, C57BL/6 mice and TXNIP-/- mice were fed with a standard diet (SD) or a high-fat and high-sugar diet (HFSD), and then we analysed biochemical and autophagy related indexes in the mice. We infected MIN6 cells with LV-TXNIP and siRNA TXNIP, then the cells were treated with free fatty acid (FFA), autophagic activator rapamycin (RAP), inhibitors of autophagy chloroquine (CQ) and bafilomycin A1(BAF), finally, we examined the changes of autophagy in MIN6 cells. The results showed that HFSD led to ß-cell dysfunction and autophagy dysregulation, which was improved by TXNIP knockout in mice. In vitro experiments, TXNIP gene silencing enhanced LC3B-I conversion to LC3B-II, reduced the protein level of P62, decreased autophagosome accumulation induced by FFA treatment, increased the glucose-stimulated insulin secretion (GSIS) and autophagic flux inhibited by treatment with CQ. TXNIP overexpression induced upregulation of LC3B-I, LC3B-II and P62, accentuating the increase in autophagy and organelle destruction induced by FFA, and exacerbated the effect of BAF on the accumulation of autophagy proteins. Increasing TXNIP levels reduced GSIS, which was reversed by treatment with RAP. In summary, our study suggested that TXNIP is a critical link between autophagy disorders and pancreatic ß-cell dysfunction.

Up-regulation of SNHG6 activates SERPINH1 expression by competitive binding to miR-139-5p to promote hepatocellular carcinoma progression.

We aimed to assess the roles of small nucleolar RNA host gene 6 (SNHG6) in hepatocellular carcinoma (HCC) progression, and establish the lncRNA-miRNA-mRNA regulation mechanism for HCC therapy. SNHG6 is one of the host genes in small nucleolar RNAs (snoRNAs), which make a difference in the development of human cancers. SERPINH1 is a gene encoding a member of the serpin superfamily of serine proteinase inhibitors with miRNA predicted by TargetScan and DIANA Tools. SNHG6, serpin family H member 1 (SERPINH1) and miR-139-5p expression levels in HCC tissues and cells were determined by quantitative real-time PCR (qRT-PCR). Migration and invasion of HCC cells were measured by transwell assay. Cell cycle analysis was determined by using flow cytometry. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and colony formation assay were performed for cell viability analysis. The expression of SERPINH1 was detected by qRT-PCR and western blot. Dual-luciferase reporter gene assay was conducted to identify the targeted relationship between miR-139-5p and SNHG6, as well as SERPINH1 and miR-139-5p. The positive regulation between SNHG6 and SERPINH1 was demonstrated in this study. In contrast, miR-139-5p was significantly down-regulated in HCC cells, the inhibition of miR-139-5p promotes the proliferation of HCC cells, and accelerated the cell cycle of HCC cells. Our study demonstrated the co-expression of SNHG6 and SERPINH1 in HCC cells for the first time, which revealed that SNHG6 could serve as a novel oncogene for the HCC therapy by its regulation.

C1QTNF1-AS1 regulates the occurrence and development of hepatocellular carcinoma by regulating miR-221-3p/SOCS3.

BACKGROUND: The aim of our study was to explore how C1QTNF1-AS1 regulated miR-221-3p/SOCS3 axis in human hepatocellular carcinoma (HCC). METHODS: Differentially expressed lncRNAs and genes were examined via RNA-seq. GO analysis and KEGG pathway enrichment analysis were carried out based on the function of dys-regulated mRNAs. RT-qPCR was employed to detect the relative mRNA expression level of C1QTNF1-AS1, miR-221-3p, SOCS3 and key genes in the JAK/STAT signaling pathway in HCC tissues and cells, and western blot analysis was conducted to detect the relative protein expression levels of SOCS3 and key proteins in the JAK/STAT signaling pathway in HCC tissues and cells. MTT assay, transwell assay and flow cytometry were utilized to assess HCC cell proliferation, invasion, migration and apoptosis. Dual luciferase reporter gene assay was used to verify the targeted relationship between C1QTNF1-AS1 and miR-221-3p, as well as between miR-221-3p and SOCS3. A tumorigenicity assay in nude mice was conducted to investigate the effects of C1QTNF1-AS1 on HCC tumor growth in vivo. RESULTS: C1QTNF1-AS1 and SOCS3 were down-regulated, while miR-221-3p was up-regulated in HCC tissues and cells. In HepG2 and Huh7 cells, overexpression of C1QTNF1-AS1 or SOCS3, as well as silence of miR-221-3p inhibited HCC cell proliferation, migration, and invasion and promoted HCC cell apoptosis. The results of the dual luciferase reporter gene assay indicated that miR-221-3p could directly target both C1QTNF1-AS1 and SOCS3. In addition, up-regulation of C1QTNF1-AS1 suppressed HCC tumor growth in vivo. CONCLUSION: Overexpression of C1QTNF1-AS1 down-regulated miR-221-3p and subsequently up-regulated SOCS3, thereby inhibiting HCC cell proliferation, migration and invasion and promoting apoptosis through the JAK/STAT signaling pathway.

LncRNA ZEB1-AS1 reduces liver cancer cell proliferation by targeting miR-365a-3p.

Liver carcinoma is one of the most common malignancies worldwide. Previous studies have demonstrated that long non-coding RNAs (lncRNAs) are crucial mediators that participate in a wide range of molecular processes associated with carcinogenesis. However, little is known about the specific mechanisms that underlie the majority of lncRNAs. Many studies have indicated that lncRNAs affect microRNA (miRNA or miR) activities via physical base-paired binding, therefore serving as competing endogenous RNAs (ceRNAs) that indirectly regulate the expression of miRNA targets. In the current study, it was revealed that lncRNA zinc-finger E-box binding homeobox 1 antisense 1 (ZEB1-AS1) serves as a ceRNA for miR-365a-3p, functioning to positively modulate E2F transcription factor 2 (E2F2) expression in liver cancer cells. Additionally, reverse transcription-quantitative polymerase chain reaction demonstrated that levels of ZEB1-AS1 were abnormally upregulated in liver cancer and this was positively correlated with E2F2 expression. Furthermore, high levels of ZEB1-AS1 exhibited a trend for poor survival in patients with liver cancer. Western blot analysis demonstrated that ZEB1-AS1 silencing could reduce E2F2 expression. EdU staining and flow cytometry analysis indicated that downregulation of ZEB1-AS1 could suppress cell proliferation and decrease the S phase proportion of liver cancer cells, which was effectively reversed by the inhibition of miR-365a-3p. ZEB1-AS1 was also determined to be physically associated with miR-365a-3p, while miR-365a-3p was revealed to target the E2F2 3'UTR for degradation or translational repression. The results also demonstrated that ZEB1-AS1 positively regulates E2F2 expression by competitively binding to miR-365a-3p. It was further revealed to enhance liver cancer cell proliferation. Thus, these results indicate that ZEB1-AS1 is required for liver cancer progression in a ceRNA dependent manner. ZEB1-AS1 may therefore be a potential target for liver cancer intervention.

miR-365 induces hepatocellular carcinoma cell apoptosis through targeting Bcl-2.

Hepatocellular carcinoma (HCC) is currently ranked as the third leading cause of cancer-related mortality worldwide. microRNAs (miRs) serve important roles in the development and progression of HCC. miR-365 has been demonstrated to function as a tumor suppressor in several types of cancer, including HCC; however, the mechanisms by which miR-365 regulates HCC apoptosis remains to be elucidated. In the present study, reverse transcription-quantitative polymerase chain reaction was performed to determine miR-365 expression levels in HCC and normal liver (LO2) cells. miR-365 overexpression was induced in SMC7721 cells using a plasmid-based system, and Cell Counting Kit-8 and TUNEL assays were performed to detect cell activity and apoptosis following miR-365 transfection. A luciferase assay was performed to determine the direct target of miR-365 in apoptosis regulation. Furthermore, a subcutaneously transplanted tumor model was established to evaluate the effects of miR-365 on tumor growth in vivo. The tumor tissue was used for further proliferation and apoptosis detection. The results of the present study indicated that miR-365 expression was significantly lower in HCC cells compared with LO2 cells (P<0.01). Transfection of SMC7721 cells with miR-365 plasmid significantly inhibited cell activity by inducing apoptosis (P<0.01). Luciferase assay indicated that miR-365 targets B-cell lymphoma 2 (Bcl-2) directly and therefore induces the downstream expression of pro-apoptotic proteins. The SMC7721 primary tumor growth was significantly reduced by miR-365 transfection (P<0.01). Further investigation demonstrated that the miR-365 group contained significantly fewer cells that were positive for proliferating cell nuclear antigen (P<0.01) and significantly more apoptotic cells (P<0.01). In conclusion, the results of the present study demonstrated that miR-365 may serve a role in inducing HCC apoptosis via directly targeting Bcl-2. This may provide a novel diagnosis and therapy target for the treatment of patients with HCC.

SATB2 induces stem-like properties and promotes epithelial-mesenchymal transition in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most prevailing malignancies, and the molecular mechanisms underlying HCC tumorigenesis remain to be further clarified. The aim of our present study was to determine the biological functions and clinical significance of SATB2 in HCC. Quantitative RT-PCR was performed to detect SATB2 mRNA expression in HCC tissues and cells. Cell proliferation, migration and invasion were investigated by MTT assay and transwell assay. Western blotting was performed to evaluate the protein expression levels. Tumor xenografts were established to explore the effects of SATB2 on HCC tumor growth in vivo. Our results indicated that SATB2 was highly expressed in HCC tissues and cell lines, and its high expression was closely correlated with aggressive clinical phenotypes and poor prognosis of HCC patients. Through gain- and loss-of function experiments, we found that SATB2 significantly promoted HCC cell proliferation, migration, and invasion in vitro and HCC tumorigenicity in vivo. Furthermore, SATB2 was identified as an important regulator of stem-like properties and epithelial to mesenchymal transition (EMT) in HCC cells. In summary, our results suggested that SATB2 is a crucial upregulator of HCC, and SATB2 might be a potential prognostic marker and a therapeutic target for HCC.

Circulation autoantibody against Lamin A/C in patients with Sjögren's syndrome.

Lamin A/C proteins are major components of nuclear laminae and were encoded by the LMNA gene. Recent studies have found that in addition to provides nuclear-membrane strength; it also regulates the gene expression. Lamin A/C has been confirmed as an autoantigen in RA, SLE and vasculitis. Anti-Lamin A/C antibodies also have been found by indirect immunofluorescence method. In this study, we used various research methods to confirm Lamin A/C is an autoantigen in Han Chinese patients with confirmed Sjögren's syndrome (SS). To further investigate the relationship between the autoimmune disease antigens, we compared the amino acid sequence of Lamin A/C epitope and several common antigens' antigenic determinant. As a result, we found that Lamin A/C has similar epitopes with U1RNP. It means that the potential relationship exist between Lamin A/C and U1RNP. Clinical data we collected also showed that anti-Lamin A/C and anti-U1RNP antibodies always appear in same serum sample. Therefore, we speculated that cross-reaction may take place between antigen and potential antigen, which have similar epitope. Then, by epitope spreading, the potential antigen can be a new autoantigen. Our study provided a new thinking for further research about the relationship between autoantigens and their development mechanism in autoimmune diseases.