Effects of miR-195-5p on ox-LDL-induced vascular endothelial cell injury through regulating FBXW7 / 中国临床药理学与治疗学

AIM: To investigate the effect of miR-195-5p on ox-LDL-induced vascular endothelial cell injury and its mechanism. METHODS: The expression of miR-195-5p in HUVECs cells induced by ox-LDL was detected by RT-PCR. Cell proliferation, the expression of oxidative stress associated factors (MDA and LDH) and apoptosis were detected by CCK-8, ELISA and flow cytometry, respectively. The potential targets of miR-195-5p were determined by dual luciferase reporter assay. The expression of target protein in ox-LDL-induced HUVECs cells and the relationship between miR-195-5p and FBXW7 expression were detected by Western blotting. RESULTS: The expression of miR-195-5p in ox-LDL-induced HUVECs cells was significantly higher than that in normal HUVECs cells. Subsequently, miR-195-5p silencing in ox-LDL-induced HUVECs cells effectively promoted the proliferation of HUVECs cells, whereas suppressed the expression of oxidative stress associated factors (MDA and LDH) and apoptosis level. Luciferase reporter assay and Western blot results showed that miR-195-5p could directly target FBXW7 protein gene and negatively regulate its expression. Finally, miR-195-5p modulated the proliferation, oxidative stress and apoptosis of HUVECs cells induced by ox-LDL by regulating the expression of FBXW7. CONCLUSION: miR-195-5p is highly expressed in HUVECs cells induced by ox-LDL. In addition, miR-195-5p can aggravate ox-LDL-induced cytotoxicity, oxidative stress and apoptosis of HUVECs cells by inhibiting the expression of FBXW7.

Serum mir-195-5p exhibits clinical significance in the diagnosis of essential hypertension with type 2 diabetes

OBJECTIVES: Diagnosis and management of essential hypertension (EH) or type 2 diabetes mellitus (T2DM) by combining comprehensive treatment and classificatory diagnosis have been continuously improved. However, understanding the pathogenesis of EH patients with concomitant T2DM and subsequent treatment remain the major challenges owing to the lack of non-invasive biomarkers and information regarding the underlying mechanisms. METHODS: Herein, we collected 200 serum samples from EH and/or T2DM patients and healthy donors (N). Gene-expression profiling was conducted to identify candidate microRNAs with clinical significance. Then, a larger cohort of the aforementioned patients and 50 N were used to identify the correlation between the tumor suppressor miR-195-5p and EH and/or T2DM. The dual-luciferase reporter assay was used to explore the target genes of miR-195-5p. The suppressive effects of miR-195-5p on the 3′-UTR of the dopamine receptor D1 (DRD1) transcript in EH patients with concomitant T2DM were verified as well. RESULTS: Compared with that in other groups, serum miR-195-5p was highly downregulated in EH patients with concomitant T2DM. miR-195-5p overexpression efficiently suppressed DRD1 expression by binding to the two 3′-UTRs. Additionally, two single nucleotide polymorphisms, including 231T-A and 233C-G, in the miR-195-5p binding sites of the DRD1 3′-UTR were further identified. Collectively, we identified the potential clinical significance of DRD1 regulation by miR-195-5p in EH patients with concomitant T2DM. CONCLUSIONS: Our data suggested that miR-195-5p circulating in the peripheral blood served as a novel biomarker and therapeutic target for EH and T2DM, which could eventually help address major challenges during the diagnosis and treatment of EH and T2DM.

MiR-195-5p targeting FGF2 inhibits malignant biological behaviors of endometrial carcinoma HEC-1B cells / 中国肿瘤生物治疗杂志

@# Objective: To explore the molecular mechanism of miR-195-5p targeting FGF2 to inhibit the proliferation, apoptosis, invasion and migration of endometrial cancer HEC-1B cells. Methods: After culture and transfection, HEC-1B cells were divided into 4 groups: HEC-1B group, miR-195-5p mimic group, pLV-FGF2 group and miR-195-5p+FGF2 group. The expressions of miR-195-5p and mRNA levels of FGF2 were detected by qRT-PCR. The targeted relationship of miR-195-5p and FGF2 was verified by luciferase assay. The protein expression of FGF2 was examined by Western blotting; Proliferation of HEC-1B cells was measured by CCK-8; Apoptosis was tested by flow cytometry; HEC-1B cell invasion was detected by transwell, and migration was measured by scratch assay. Results: Compared with HEC-1B group, the expression of miR-195-5p in miR-195-5p mimic group was elevated while FGF2 mRNA level was declined (all P<0.01). Luciferase assay indicated that FGF2 was a target of miR-195-5p. Compared with HEC-1B group, the protein level of FGF2 in miR-195-5p mimic group was decreased, and the protein levels of FGF2 in pLV-FGF2 group were enhanced (P<0.01). The protein levels of FGF2 in miR-195-5p+FGF2 group were lower than that in pLV-FGF2 group (all P<0.01). The proliferation in miR-195-5p mimic group was lower than HEC-1B group (P<0.01), while the proliferation in pLV-FGF2 group was higher than that in HEC-1B group (all P<0.01). Compared with HEC-1B group, apoptosis in miR-195-5p mimic group was increased, and apoptosis in pLV-FGF2 group was decreased (P<0.01); moreover, apoptosis in miR-195-5p+FGF2 group was higher than that in pLV-FGF2 group (P<0.01). Compared with HEC-1B group, the number of invasive cells per field and the rate of wound healing in miR195-5p mimic group were decreased, while those in pLV-FGF2 group was enhanced (P<0.01); moreover, the number of invasive cells per field and the rate of wound healing in miR-195-5p+FGF2 group was lower than those in pLV-FGF2 group (all P<0.01). Conclusion: miR-195-5p inhibits proliferation, invasion and migration and promotes apoptosis of endometrial cancer HEC-1B cells by targeting FGF2, and could be used as a treatment target of endometrial cancer.