UBC Mediated by SEPT6 Inhibited the Progression of Prostate Cancer

Background. Prostate cancer is one of the most common malignancies in men. Protein ubiquitination is an important mechanism for regulating protein activity and level in vivo. We aimed to study the mechanism of SEPT6 and UBC action in prostate cancer to identify new targets. Methods. The ubiquitin-protein and the ubiquitin coding gene UBA52, UBA80, UBB, and UBC expressions were detected in clinical tissues and cells. Overexpression and knockdown of UBC were performed in prostate cancer DU145 cells. Cell Counting Kit 8 (CCK-8) assay was performed to detect cell proliferation. Cell cycle at 24 h was detected by flow cytometry. Clonal formation assay was used to measure cell clone number. Immunofluorescence (IF) was performed to detect the colocalization of SEPT6 and UBC in prostate cancer cells. Next, we overexpressed or knocked down SEPT6 expression in DU145 cells. Pearson correlation coefficient was applied to analyze the relationship between SEPT6 and UBC in prostate cancer tissue. oe-SEPT6+oe-UBC coexpressing cells were constructed to detect the upstream and downstream relationship between SEPT6 and UBC on prostate cancer cells. The tumor formation experiment was performed to explore SEPT6/UBC effect on prostate cancer. Results. UBC was upregulated in prostate cancer tissues and cells. Overexpression of UBC promoted cell survival and proliferation. IF revealed the colocalization of SEPT6 and UBC in prostate cancer cells. UBC expression decreased after oe-SEPT6, while increased after sh-SEPT6, indicating that UBC was downstream of SEPT6. Pearson correlation coefficient analysis showed that SEPT6 was negatively correlated with UBC in prostate cancer tissues. SEPT6 as an upstream gene of UBC regulated prostate cancer cell behavior through UBC. The tumor formation experiment showed that SEPT6 could inhibit tumor growth. Conclusion. In general, SEPT6 inhibited UBC expression, thereby reducing the overall ubiquitination level, affecting the expression level of downstream cell proliferation-related genes, and then affecting the progression of prostate cancer.

Bedrock gorge incision via anthropogenic meander cutoff

Bedrock river-gorge incision represents a fundamental landscape-shaping process, but a dearth of observational data at >10 yr timescales impedes understanding of gorge formation. I quantify 102 yr rates and processes of gorge incision using historical records, field observations, and topographic and image analysis of a human-caused bedrock meander cutoff along the North Fork Fortymile River in Alaska (USA). Miners cut off the meander in 1900 CE, abruptly lowering local base level by 6 m and forcing narrowing and steepening of the channel across a knickpoint that rapidly incised upstream. Tectonic quiescence, consistent rock erosivity, and low millennial erosion rates provide ideal boundary conditions for this 102 yr gorge-formation experiment. Initial fast knickpoint propagation (23 m/yr; 1900–1903 CE) slowed (4 m/yr; 1903–1981 CE) to diffusion (1981–2019 CE) as knickpoint slope decreased, yielding an ~350-m-long, 6-m-deep gorge within the pre–1900 CE channel. Today, diffusion dominates incision of a 500-m-long knickzone upstream of the gorge, where sediment transport likely limits ongoing adjustments to the anthropogenic cutoff. Results elucidate channel width, slope, discharge, and sediment dynamics consistent with a gradual transition from detachment- to transport-limited incision in fluvial adjustment to local base-level lowering.

LRP16 Promotes Proliferation and Migration of Esophageal Squamous Cell Carcinoma by Regulating Hippo Signaling Pathway

The present study aimed to investigate the expression of LRP16 in the development of ESCC and the relationship between Hippo signaling pathway and LRP16. Immunohistochemistry was used to detect the expression of LRP16 in ESCC tissues. After transfection, the expression of LRP16 was detected by reverse transcription quantitative PCR (RT-qPCR) and western blot techniques. Cell counting kit (CCK-8), clone formation experiment, flow cytometry and wound healing were used to determine the proliferation, apoptosis, cell cycle and migration of ESCC cells. The changes of factors related to Hippo signaling pathway were determined via RT-qPCR and western blot experiments. The results showed that the LRP16 expression in ESCC tissues was higher than that in normal tissues. High expression of LRP16 was related to the depth of invasion, TNM stage and lymph node metastasis of ESCC. Furthermore, the knockdown of LRP16 inhibited proliferation, migration and promoted cell apoptosis and made cells arrested in G2/M phase. It also resulted in decreased expression of Yes-associated protein (YAP), and increased expression of mammalian STE20-like protein kinase (MST1/2), suggesting that LRP16 promoted the development of ESCC through Hippo signaling pathway. The results of this study suggest that LRP16 may be a carcinogenic gene of ESCC and promotes the progression of ESCC through the regulation of Hippo signaling pathway. Our study provides a new idea for the diagnosis and treatment of ESCC in the future.

Deubiquitinating enzyme USP48 mediates pyroptosis and meliorates immune evasion in tumors by stabilizing GSDME expression

Pyroptosis is a kind of programmed cell death, which is characterized by the activation of inflammatory caspase and the cleavage of gasdermin proteins. It is widely involved in the occurrence and development of tumors. Studies have shown that ubiquitin related proteins play a key regulatory role in many biological processes. However, the role and molecular mechanism of ubiquitin-related proteins in pyroptosis have not been well identified. Here, using CRISPR-Cas9 screening, we identified a deubiquitinating enzyme (USP48) that has the most significant regulatory effect on cell pyroptosis. USP48 stabilizes GsderminE (GSDME) expression by causing deubiquitination of it, thereby achieving its regulatory effect on pyroptosis. USP48 prevents the degradation of GSDME by inhibiting K63-linked ubiquitination at positions K120 and K189 of GSDME. Clinical tissue testing confirmed that the expression of USP48 has a significant positive correlation with GSDME and pyroptosis-related factors. GSDME plays a crucial role in the regulation of cell pyroptosis by USP48. The single-cell sequencing results showed that the functions of T cells and tumor-associated macrophages in the tumor microenvironment are inhibited to varying degrees after USP48 gene knockout. Finally, the tumor formation experiment in mice confirmed that overexpression of USP48 could effectively improve the therapeutic effect of PD-1 inhibitors. These findings define a pyroptosis regulation pathway and indicate that activation of the pharmacological activity of USP48 may provide an effective strategy to sensitize cancer cells to pyroptosis-related immunotherapeutic resistance.

miR-3156-5p Enhances Hypoxia-Induced Angiogenesis in Hepatocellular Carcinoma via Modulating the SOCS5/HIF-1α/VEGF Pathway

Background: MicroRNAs (miRNAs) are dysregulated in hypoxia-induced hepatocellular carcinoma (HCC). This study probed the regulatory mechanism of miR-3156-5p on HCC under hypoxia. Methods: HCC cells (HepG2) were exposed to normoxia or hypoxia, and the conditioned medium (CM) of HepG2 was applied. Quantitative reverse transcription PCR (qRT-PCR) was implemented to analyze the miR-3156-5p profile. The cell counting kit-8 (CCK-8) assay and the colony formation experiment were conducted to measure cell proliferation, colony formation, and angiogenesis. Results: The results manifested that miR-3156-5p was up-regulated in HCC cells and endothelial cells under hypoxia, and up-regulating miR-3156-5p boosted HCC cell proliferation, endothelial cell angiogenesis, and HIF-1α/VEGF expression. Conclusions: miR-3156-5p activates the HIF-1α/VEGF pathway by hampering SOCS5, thereby enhancing the angiogenic potential of hypoxia-induced endothelial cells in HCC cells.

Metformin exerts anti-AR-negative prostate cancer activity via AMPK/autophagy signaling pathway

Background Encouraged by the goal of developing an effective treatment strategy for prostate cancer, this study explored the mechanism involved in metformin-mediated inhibition of AR-negative prostate cancer. Methods Cell behaviors of DU145 and PC3 cells were determined by CCK8 test, colony formation experiment and scratch test. Flow cytometry was used to detect cell cycle distribution. Cell autophagy was induced with metformin, and an autophagy inhibitor, 3-MA, was used to assess the level of autophagy. Detection of LC3B by immunofluorescence was conducted to determine autophagy level. Cell proliferation, autophagy and cell cycle were examined by performing Western blot. DU145 and PC3 cell lines were transfected with AMPK siRNA targeting AMPK-α1 and AMPK-α2. Tumor formation experiment was carried out to evaluate the anti-prostate cancer effect of metformin in vivo. Results The inhibitory effect of metformin on the proliferation of prostate cancer cell lines was confirmed in this study, and the mechanism of such an effect was related to autophagy and the block of cell cycle at G0/G1 phase. Metformin also induced the activation of AMPK, markedly promoted expression of LC3II, and down-regulated the expression of p62/SQSTM1. Animal experiments showed that the tumor volume of metformin group was smaller, meanwhile, the levels of p-AMPK (Thr172) and LC3B were up-regulated and the Ki-67 level was down-regulated, without abnormalities in biochemical indicators. Conclusion This study found that autophagy induction might be the mechanism through which metformin suppressed the growth of AR-negative prostate cancer. Moreover, the activation of AMPK/autophagy pathway might be a therapeutically effective for treating AR-negative prostate cancer in the future.

miR-29a-3p inhibits endometrial cancer cell proliferation, migration and invasion by targeting VEGFA/CD C42/PAK1

Background This study aimed to investigate the mechanism of miR-29a-3p in regulating endometrial cancer (EC) progression. Methods A total of 72 EC patients were enrolled. EC cells were transfected. Cells proliferation, cloning ability, migration and invasion were researched by MTT assay, colony formation experiment, cell scratch test and Transwell experiment respectively. Dual-luciferase reporter assay was performed. Xenograft experiment was conducted using nude mice. miR-29a-3p, VEGFA, CDC42, PAK1 and p-PAK1 expression in cells/tissues was investigated by qRT-PCR and Western blot. Results miR-29a-3p expression was aberrantly reduced in EC patients, which was associated with poor outcome. miR-29a-3p inhibited EC cells proliferation, cloning formation, migration and invasion (P < 0.05 or P < 0.01 or P < 0.001). miR-29a-3p inhibited CDC42/PAK1 signaling pathway activity in EC cells (P < 0.01). VEGFA expression was directly inhibited by miR-29a-3p. miR-29a-3p suppressed EC cells malignant phenotype in vitro and growth in vivo by targeting VEGFA/CDC42/PAK1 signaling pathway (P < 0.05 or P < 0.01). Conclusion miR-29a-3p inhibits EC cells proliferation, migration and invasion by targeting VEGFA/CDC42/PAK1 signaling pathway.

TSP1 is the essential domain of SEMA5A involved in pannus formation in rheumatoid arthritis

Objective In this study, we explored the effect of semaphorin5A (SEMA5A) on rheumatoid arthritis (RA) pathogenesis and its specific TSP1 domain on pannus formation. Methods The expression of SEMA5A was detected in synovium, fibroblast-like synoviocytes (FLS) and synovial fluid of RA patients and healthy controls (HCs) by q-PCR, IHC, WB and ELISA. SEMA5A-mAb intervention was performed to appraise the severity of joints in CIA model. Transcriptome sequencing and bioinformatics analysis in SEMA5A transfected FLS from HCs were performed to screen differentially expressed genes after SEMA5A overexpression. MTT assay in RA-FLS, chicken embryo allantoic membrane experiment and tube formation experiment were used to clarify the influence of SEMA5A on cell proliferation and angiogenesis. Furthermore, rescue experiment verified the function of TSP1 domain of SEMA5A in the progress of RA with Sema5a-/- CIA mice. Results The expression of SEMA5A increased in RA compared with HCs. Simultaneously, SEMA5A-mAb significantly attenuated joint injury and inflammatory response in CIA models. Besides, transcriptome sequencing and angiogenesis-related experiments verified the ability of SEMA5A to promote FLS proliferation and angiogenesis. Moreover, TSP1 was proved as an essential domain in SEMA5A-inducing angiogenesis in vitro. Additionally, rescue of TSP1-deleted SEMA5A failed to deteriorate the severity of arthritis in CIA model constructed with Sema5a -/- mice. Conclusions In summary, up-regulation of SEMA5A was firstly confirmed in pathological lesion of RA patients. Furthermore, the treatment of SEMA5A-mAb attenuated the progress of RA in CIA model. Moreover, TSP1 was indicated as the key domain of SEMA5A to promote pannus formation in RA.

On possibilities for development of human adaptive resources in arctic conditions

This article presents the results of an experimental study aimed to investigate perspectives of the intentional development of adaptive resources of the human organism in the context of the harsh Arctic environmental conditions. The effectiveness of the author’s comprehensive psychophysical training program designed to develop mechanisms of human psycho-physiological and mental self-regulation has been tested in a formative experiment. The study was conducted on a sample of 18 clinically healthy female patients with an average age of 40±5.6 years and the northern experience of 18.3 ± 10.6 years. The results showed a significant change in several measurements indicating the improvement in some important characteristics of the functional state of subjects as a result of the psychophysical training. These changes suggest an increase in the adaptive potential of the subjects due to the formation experiment.