HCMV-miR-US33-5p promotes apoptosis of aortic vascular smooth muscle cells by targeting EPAS1/SLC3A2 pathway.

BACKGROUND: In patients with acute aortic dissection (AAD), increased vascular smooth muscle cell (VSMC) apoptosis has been found. Human cytomegalovirus (HCMV)-miR-US33-5p was significantly increased in the plasma of patients with AAD. However, the roles of miR-US33-5p in human aortic VSMC (HA-VSMC) apoptosis remain to be elucidated. METHODS: In the current study, cell apoptosis was analyzed by flow cytometry, cell proliferation by CCK-8 assay, and differentially expressed genes by RNA sequencing. Luciferase reporter assay was used for binding analysis between miR-US33-5p and endothelial PAS domain protein 1 (EPAS1), and EPAS1 and amino acid transporter heavy chain, member 2 (SLC3A2). The enrichment degree of SLC3A2 promoter DNA was analyzed by chromatin immunoprecipitation assay. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and immunoblotting were performed for measuring messenger RNA (mRNA) and protein levels, respectively. RESULTS: It was found that HCMV infection inhibited proliferation but promoted HA-VSMC apoptosis by upregulating HCMV-miR-US33-5p. Transfection of HCMV-miR-US33-5p mimics the significant effect on several signaling pathways including integrin signaling as shown in the RNA sequencing data. Western blotting analysis confirmed that HCMV-miR-US33-5p mimics suppression of the activity of key factors of the integrin signal pathway including FAK, AKT, CAS, and Rac. Mechanistic study showed that HCMV-miR-US33-5p bound to the 3'-untranslated region of EPAS1 to suppress its expression, leading to suppression of SLC3A2 expression, which ultimately promoted cell apoptosis and inhibited cell proliferation. This was confirmed by the findings that silencing EPAS1 significantly reduced the SLC3A2 expression and inhibited proliferation and key factors of integrin signal pathway. CONCLUSIONS: HCMV-miR-US33-5p suppressed proliferation, key factors of integrin signal pathway, and EPAS1/SLC3A2 expression, but promoted HA-VSMC apoptosis. These findings highlighted the importance of HCMV-miR-US33-5p/EPAS1/SCL3A2 signaling and may provide new insights into therapeutic strategies for AAD.

miR-876-3p suppresses the progression of colon cancer and correlates the prognosis of patients.

BACKGROUND: miR-876-3p has been identified to be downregulated in colon cancer, implying the potential biological function in the progression and prognosis of colon cancer. The clinical significance and the biological function of miR-876-3p were investigated in this study to assess the potential of miR-876-3p in acting as a novel biomarker of the progression of colon cancer. METHODS: The expression of miR-876-3p in colon cancer was evaluated by RT-qPCR. The clinical significance of miR-876-3p was assessed by associated its expression level with the clinical features and prognosis of patients. The biological function of miR-876-3p was estimated by the CCK8 and Transwell assay in vitro. RESULTS: The significant downregulation of miR-876-3p was observed in colon cancer tissues and cells, which was closely associated with the lymph node metastasis status, TNM stage, and the perineural invasion of patients. miR-876-3p served as an independent indicator that was negatively associated with the prognosis of patients. In colon cancer cells, miR-876-3p showed significant inhibitory effects on cell proliferation, migration, and invasion, indicating its tumor suppressor role in the progression of colon cancer. CONCLUSION: miR-876-3p might be involved in colon cancer development, which provides a potential therapeutic target for colon cancer treatment.

Long Noncoding RNA NORAD Promotes Fracture Healing through Interacting with Osteoblast Differentiation via Targeting miR-26a.

The present study was designed to evaluate the dynamic expression of lncRNA NORAD in fracture healing of patients with brittle fractures and explore the function and mechanism of NORAD in regulating osteoblastic proliferation, differentiation, and apoptosis. The expression level of NORAD was detected by quantitative real-time PCR. The proliferation, differentiation, and apoptosis of osteoblasts were analyzed by MTT assay, ELISA, and flow cytometry. Luciferase report analysis was used to confirm the interaction between NORAD and its target ceRNA miR-26a. This study showed no significant differences in serum NORAD expression on the 7th day during fracture healing in patients, but increased expression of NORAD was certified on the 14, 21, and 28 days after fixation. Overexpression of NORAD promoted the proliferation and differentiation of osteoblasts and suppressed the apoptosis of osteoblasts. miR-26a proved to be the target gene of NORAD and was inhibited by overexpression of NORAD in osteoblasts. The enhanced expression of miR-26a was negatively linked to the lessened expression of NORAD. NORAD could accelerate the proliferation and differentiation of osteoblasts and inhibit apoptosis, thereby promoting fracture healing.

MicroRNA-455-3p promotes osteoblast differentiation via targeting HDAC2.

BACKGROUND: Fragility fracture commonly occurs in the elderly, the basis of fracture healing is osteoblast regeneration. The study measured the expression changes of microRNA-455-3p during fracture healing in patients with fragility fractures, and explored its influence on osteoblast differentiation. METHODS: 108 postmenopausal women with osteoporosis were recruited, in which 58 cases with fragility fracture. qRT-PCR was used for the measurement of miR-455-3p levels. MC3T3-E1 cells were induced differentiation by BMP-2. ELISA was performed for the measurement of alkaline phosphates (ALP), runt-related transcription factor-2 (RUNX2), osteocalcin (OCN), and Collagen I. Luciferase reporter gene assay was done for the target gene analysis. RESULTS: Serum miR-455-3p was significantly decreased in both osteoporosis and fragility fracture patients compared with the control group, which was most deficient in patients with fragility fracture. With the extension of treatment time, the level of miR-455-3p in serum increased gradually and reached the highest level at 4 weeks of treatment. Levels of miR-455-3p continued to increase on the 7th and 14th days after induction of cell differentiation. MiR-455-3p overexpression promoted cell proliferation, and increased the levels of osteoblast differentiation markers, including ALP, OCN, Collagen I, and RUNX2. MiR-455-3p in MC3T3-E1 cells was directly bound to HDAC2 and negatively regulated. Both MC3T3-E1 differentiation and the fracture healing of patients were accompanied by progressively reduced HDAC2. CONCLUSIONS: MiR-455-3p promotes osteogenic differentiation which may be associated with fracture healing, HDAC2 acts as a target of miR-455-3p in the underlying mechanism.

Quality evaluation of HPV vaccine-related online messages in China: a cross-sectional study.

Since 2019, three types of HPV vaccine have been approved for use in mainland China. High quality messages are crucial for vaccine acceptance, but little is known about the online information quality concerning HPV vaccine in China. "HPV vaccine" and "cervical cancer vaccine" in the form of Chinese were used as keywords through search engines from personal computer (PC), portable mobile device (PMD), and WeChat Public Accounts in 2019. Readability, information content, as well as DISCERN scores were evaluated for each message included. Characteristics associated with quality indicators of the messages were also analyzed. A total of 294 messages from PC engines (104, 35%), PMD engines (128, 43%) and WeChat (62, 21%) were assessed. Most (269, 91%) messages required at least undergraduate readability level. The most frequently reported theme was HPV vaccine and its function (273, 93%), while the least was information regarding quality, safety and side effects (129, 44%). The frequency of messages with at least one error was 132 (45%). The median of sum DISCERN scores was 42 (IQR = 14), and only one (< 1%) message showed good DISCERN quality. Messages retrieved from PC engines and those with pictures were of better overall quality. The overall quality of HPV vaccine-related online messages in Chinese websites was not optimal. Government and health professionals should promote information quality construction to improve the status of HPV vaccination messages online.

Enhanced anti-angiogenic effect of a deletion mutant of plasminogen kringle 5 on neovascularization.

Kringle 5 (K5), a proteolytic fragment of plasminogen, has been proved to be an angiogenic inhibitor. Previously, we have evaluated the effect of K5 on the vascular leakage and neovascularization in a rat model of oxygen-induced retinopathy. In this study, we expressed K5 and a deletion mutant of K5 (K5 mutant) in a prokaryocyte expression system and purified them by affinity chromatography. K5 mutant was generated by deleting 11 amino acids from K5 while retaining the three disulfide bonds. The anti-angiogenic activity of intact K5 and K5 mutant were compared in endothelial cells and retinal neovascularization rat model. K5 mutant inhibited the proliferation of primary human retinal capillary endothelial cells (HRCEC) in a concentration-dependent manner, with an apparent EC50 of approximate 35 nmol/L, which is twofold more potent than intact K5. In the even higher concentration range, K5 mutant did not inhibit pericytes from the same origin of HRCEC, which suggested an endothelial cell-specific inhibition. K5 mutant had no effect on normal liver cells and Bel7402 hepatoma cells even at high concentration range either. Intravitreal injection of the K5 and mutant in the oxygen-induced retinopathy rat model both resulted in significantly fewer neovascular tufts and nonperfusion area than controls with PBS injection, as shown by fluorescein angiography. Furthermore, K5 mutant exhibited more strong inhibition effect on neovascularization than intact K5 by quantification of vascular cells. These results suggest that this K5 deletion mutant is a more potent angiogenic inhibitor than intact K5 and may have therapeutic potential in the treatment of those disorders with neovascularization, such as solid tumor, diabetic retinopathy, age-related macular degeneration, rheumatoid arthritis, and hyperplasia of prostate.