KLHL21, a novel gene that contributes to the progression of hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) has very high prevalence and associated-mortality. However, targeted therapies that are currently used in clinical practice for HCC have certain limitations, in part because of the lack of reliable and clinically applicable biomarkers that can be used for diagnosis and prognosis assessments and for the surveillance of treatment effectiveness. METHODS: Meta-analysis was used to analyze the integrated microarray data for global identification of a set of robust biomarkers for HCC. Quantitative RT-PCR (qRT-PCR) was performed to validate the expression levels of selected genes. Gene expression was inhibited by siRNA. CellTiter 96® AQueous One Solution Cell Proliferation assays were used to determine cell proliferation, and Transwell assays were used to determine cell migration and invasion potential. RESULTS: Meta-analysis of the expression data provided a gene expression signature from a total of 1525 patients with HCC, showing 1529 up-regulated genes and 478 down-regulated genes in cancer samples. The expression levels of genes having strong clinical significance were validated by qRT-PCR using primary HCC tissues and the paired adjacent noncancerous liver tissues. Up-regulation of VPS45, WIPI1, TTC1, IGBP1 and KLHL21 genes and down-regulation of FCGRT gene were confirmed in clinical HCC samples. KLHL21 was the most promising gene for potential use as a bioclinical marker in this analysis. Abrogating expression of it significantly inhibited cell proliferation, migration and invasion. CONCLUSIONS: Our study suggests that KLHL21 is a potential target for therapeutic intervention. Our findings also provide novel candidate genes on a genome-wide scale, which may have significant impact on the design and execution of effective therapy of HCC patients.

Effects of short-chain fatty acids in inhibiting HDAC and activating p38 MAPK are critical for promoting B10 cell generation and function.

B10 cells are regulatory B cells capable of producing IL-10 for maintaining immune homeostasis. Dysregulation of B10 cells occurs in autoimmune and inflammatory diseases. Modulation or adoptive transfer of B10 cells is a promising therapeutic strategy. The short-chain fatty acids (SCFAs), the metabolites of microbiota, play a critical role in maintaining immune homeostasis and are the potential drugs for the modulation of B10 cells. It is not clear whether and how SCFAs upregulate the frequency of B10 cells. Here, we found that SCFAs could promote murine and human B10 cell generation in vitro. Upregulation of B10 cells by butyrate or pentanoate was also observed in either healthy mice, mice with dextran sodium sulfate (DSS)-induced colitis, or mice with collagen-induced arthritis. Moreover, SCFA treatment could ameliorate clinical scores of colitis and arthritis. Adoptive transfer of B cells pretreated with butyrate showed more alleviation of DSS-induced colitis than those without butyrate. A further study demonstrates that SCFAs upregulate B10 cells in a manner dependent on their histone deacetylase (HDAC) inhibitory activity and independent of the G-protein-coupled receptor pathway. Transcriptomic analysis indicated that the MAPK signaling pathway was enriched in B10 cells treated with butyrate. A study with inhibitors of ERK, JNK, and p38 MAPK demonstrated that activating p38 MAPK by butyrate is critical for the upregulation of B10 cells. Moreover, HDAC inhibitor has similar effects on B10 cells. Our study sheds light on the mechanism underlying B10 cell differentiation and function and provides a potential therapeutic strategy with SCFAs and HDAC inhibitors for inflammation and autoimmune diseases.

Porphyromonas gingivalis induces periodontitis, causes immune imbalance, and promotes rheumatoid arthritis.

Periodontitis induced by bacteria especially Porphyromonas gingivalis (P. gingivalis) is the most prevalent microbial disease worldwide and is a significant risk factor for systemic diseases such as rheumatoid arthritis (RA). RA and periodontitis share similar clinical and pathologic features. Moreover, the prevalence of RA is much higher in patients with periodontitis than in those without periodontitis. To explore the immunologic mechanism of periodontitis involved in RA, we established a mouse model of periodontitis and then induced RA. According to the results of paw thickness, arthritis clinical score, arthritis incidence, microscopic lesion using H&E staining, and micro-CT analysis, periodontitis induced by P. gingivalis promoted the occurrence and development of collagen-induced arthritis (CIA) in mice. Furthermore, periodontitis enhanced the frequency of CD19+ B cells, Th17, Treg, gMDSCs, and mMDSCs, whereas down-regulated IL-10 producing regulatory B cells (B10) in CIA mice preinduced for periodontitis with P. gingivalis. In vitro stimulation with splenic cells revealed that P. gingivalis directly enhanced differentiation of Th17, Treg, and mMDSCs but inhibited the process of B cell differentiation into B10 cells. Considering that adoptive transfer of B10 cells prevent RA development, our study, although preliminary, suggests that down-regulation of B10 cells may be the key mechanism that periodontitis promotes RA as the other main immune suppressive cells such as Treg and MDSCs are up-regulated other than down-regulated in group of P. gingivalis plus CIA.

Clinical therapeutic efficacy of mesenchymal stem cells derived from adipose or bone marrow for knee osteoarthritis: a meta-analysis of randomized controlled trials.

Aim: This meta-analysis, only including randomized controlled trials (RCTs), was conducted to assess separately and compare the therapeutic efficacy of adipose-derived mesenchymal stem cells (ADMSCs) and bone marrow-derived mesenchymal stem cells (BMSCs) for knee osteoarthritis (OA) at the same follow-up time. Methods: Potential relevant researches were identified from PubMed, Web of Science, Embase, Cochrane Library and clinicaltrials.gov. The data, from clinical trials concentrating on knee OA treated with ADMSCs or BMSCs, were extracted and pooled for meta-analysis to compare the clinical outcomes of patients with knee OA in visual analog scale (VAS), Western Ontario McMaster Universities Osteoarthritis Index (WOMAC), Lysholm knee scale (Lysholm) and Tegner activity scale (Tegner). Results: Nine randomized controlled trials including a total of 377 patients met the inclusion criteria. This meta-analysis obtained the following results. First, the improvement of VAS scores was statistically significant after BMSCs treatment at 6-, 12- and 24-month follow-up compared with control groups (p < 0.01). In contrast, the improvement of WOMAC scores was of no statistical significance, but showed a positive trend with the prolongation of the follow-up time (6 months: mean difference [MD] = 6.51; 95% CI: -2.38 to 15.40; p = 0.15; 12 months: MD = -6.81; 95% CI: -13.94 to 0.33; p = 0.06). Lysholm scores presented a similar pattern (12 months: MD = 1.93; 95% CI: -11.52 to 15.38; p = 0.78; 24 months: MD = 8.94; 95% CI: 1.45 to 16.43; p = 0.02). Second, VAS and WOMAC scores of patients after ADMSCs treatment were significantly improved at any follow-up time (p ≤ 0.05). The improvement of Lysholm scores was of no statistical significance compared with control groups, although treatment outcome at 12-month follow-up was better than that at 24-month follow-up, which was debatable because only data of one clinical trial were pooled in the analysis (12 months: MD = 7.50; 95% CI: -1.94 to 16.94; p = 0.12; 24 months: MD = 5.10; 95% CI: -3.02 to 13.22; p = 0.22). Finally, by comparing the statistical results of VAS and WOMAC scores, it could be concluded that the therapeutic effect of ADMSCs on knee OA was more effective than that of BMSCs. Conclusion: This meta-analysis showed that regeneration with BMSCs or ADMSCs had a great application potential in the treatment of patients with knee OA, and ADMSCs tended to be superior to BMSCs according to the limited clinical evidences available.

Expression and Function of Tetraspanins and Their Interacting Partners in B Cells.

Tetraspanins are transmembrane proteins that modulate multiple diverse biological processes, including signal transduction, cell-cell communication, immunoregulation, tumorigenesis, cell adhesion, migration, and growth and differentiation. Here, we provide a systematic review of the involvement of tetraspanins and their partners in the regulation and function of B cells, including mechanisms associated with antigen presentation, antibody production, cytokine secretion, co-stimulator expression, and immunosuppression. Finally, we direct our focus to the signaling mechanisms, evolutionary conservation aspects, expression, and potential therapeutic strategies that could be based on tetraspanins and their interacting partners.

Gene expression profiling and functional analysis reveals that p53 pathway-related gene expression is highly activated in cancer cells treated by cold atmospheric plasma-activated medium.

BACKGROUND: Cold atmospheric-pressure plasma (CAP) has been considered a promising strategy for anti-cancer treatment. Traditionally, CAP was employed to kill cancer cells or tumor tissues by direct irradiation. However, CAP has some disadvantages such as infiltration capacity and storage convenience. Recently, plasma-activated medium (PAM) was used as an alternative strategy to treat cancer cells or tumors. The novel PAM approach has potential as an anti-cancer therapy. OBJECTIVE: To reveal the global activation of signaling pathways in oral cancer cells induced by PAM. METHODS: Oral squamous cell line SCC15 were treated by PAM and gene expression profiles were evaluated by using RNA-seq. Functional analyses were employed to reveal the global responses of SCC15 cells with PAM stimulation. QRT-PCR and Western blot were carried out to validate the expression levels of selected genes. RESULTS: More than 6G clean data per sample were obtained in PAM-treated SCC15 cells. A total of 934 differentially expressed genes (DEGs) were identified and GO analysis implicated the deep involvement of biological process. KEGG mapping further clustered 40 pathways, revealing that "p53 pathway" was significantly enriched. SCC15 cells were commonly used as a p53-null cell line. Therefore, the enriched p53 pathway-related genes in our analysis might be activated by other stimulators, in a p53-independent manner. Gene set enrichment analysis (GSEA) was also performed to evaluate changes at the gene-sets level. The results demonstrated not only the high engagement of "p53 pathway" but also the involvement of novel pathways such as hypoxia pathway. CONCLUSIONS: The present study elucidates the transcriptomic changes of PAM treated SCC15 cells, containing highly enriched DEGs involved in "p53 pathway". Our analysis in this work not only provides genomic resources for future studies but also gives novel insights to uncover the molecular mechanism of PAM stimulation.