Effect of LINC00707 on proliferation, migration and inflammatory factors of human vascular smooth muscle cells induced by ox-LDL by targeting miR-30c-5p / 中国药理学通报

Aim To explore the effect of LINC00707 on the proliferation, migration and inflammatory factors of human vascular smooth muscle cells induced by oixidized low-density lipoprotein (ox-LDL) and its possible mechanism. Methods ox-LDL was used to induce human vascular smooth muscle cells (HVSMCs) to establish an atherosclerotic cell model. si-NC, siLINC00707, miR-NC, miR-30c-5p mimic were trans-fected into HVSMCs, and then 100 mg • L~ ox-LDL was added to the cells. si-LINC00707 and anti-miRNC, or si-LINC00707 and miR-30c-5p Inhibitor were co-transfected into HVSMCs and then treated with 100 mg • L ox-LDL. qRT-PCR was used to detect the expression of LINC00707 and miR-30c-5p. CCK-8 and Transwell test were used to detect cell proliferation and migration. ELISA was used to detect the levels of IL-6, TNF-a, and IL-10. The dual-luciferase reporter experiment was used to detect the targeting relationship between LINC00707 and miR-30c-5p. Western blot was used to detect the protein expression of E-cadherin and N-cadherin. Results The expression of LINC00707 in HVSMCs induced by ox-LDL increased (P <0. 05), while the expression of miR-30c-5p decreased (P < 0. 05). After transfection with siLINC00707 or miR-30c-5p mimic, cell viability, the protein level of N-cadherin, the levels of IL-6 and TNF-a decreased (P < 0. 05), and the number of im grating cells decreased (P<0. 05), while the protein level of E-coadherin and the level of IL-10 increased (P <0. 05). LINC00707 could target miR-30c-5p. After co-transfection with si-LINC00707 and miR-30c-5p inhibitor, cell survival rate, the protein level of N-cadherin, the levels of IL-6 and TNF-α increased (P < 0.05), and the number of migrating cells increased (P <0. 05), while the protein level of E-cadherin and the level of IL-10 decreased (P < 0. 05). Conclusion Down-regulation of the expression of LINC00707 could inhibit the proliferation, migration and inflammation of human vascular smooth muscle cells induced by ox-LDL by promoting the expression of miR-30c-5p.

Effect of miR-30c targeting Wnt/β-catenin signal on the proliferation and apoptosis of human retinal endothelial cells induced by high glucose / 中华内分泌外科杂志

Objective:To investigate the protective effect and mechanism of miR-30c targeting Wnt/β-catenin signal on the proliferation and apoptosis of human retinal endothelial cells (HRECs) induced by high glucose.Methods:Human retinal endothelial cells (HRECs) were cultured and given normal concentration (control group) and high concentration glucose (high glucose group) respectively. According to the experimental design, miR-30c mimic, negative control (miR-NC), Wnt1 overexpression vector (pcDNA3.1-Wnt1) and empty vector (pcDNA3.1) were transfected respectively. RT-PCR was used to detect the expression level of miR-30c in each group. Western blot was used to detect the expression levels of Wnt1, β-catenin and GSK-3β protein in each group. The dual luciferase experiment verified the targeting relationship of miR-30c to Wnt1. Thiazole blue method was used to detect the proliferation activity of each group. Flow cytometry was employed to detect the level of apoptosis of each group of cells.Results:Compared with the control group, the expression of miR-30c in the high glucose group was significantly reduced [ (0.94±0.11) vs (0.32±0.06), P<0.001]; compared with the control group, the cell proliferation activity of the high glucose group was significantly reduced, and the apoptosis rate was significantly increased [ (0.75±0.08) vs (0.13±0.04), (3.53±0.29) % vs (14.89±0.94) %, P<0.001]; compared with the high glucose+miR-NC group, the cell proliferation activity of the high glucose+miR-30c group was significantly increased, and the apoptosis rate was reduced [ (0.14±0.04) vs (0.64±0.06), (14.14±0.86) % vs (6.28±0.45) %, P<0.001]; compared with the miR-NC group, the luciferase activity of the miR-30c group co-transfected with WT-Wnt1 was significantly reduced [ (0.97±0.09) vs (0.26±0.03), P<0.001]; compared with the control group, the protein expression of Wnt1, β-catenin and GSK-3β in the high glucose group were significantly increased [ (0.43±0.05) vs (1.02±0.09), (0.25±0.04) vs (0.82±0.10), (0.39±0.04) vs (0.76±0.11), P<0.001]; compared with the high glucose+miR-NC group, the protein expression of Wnt1, β-catenin and GSK-3β in the high glucose+miR-30c group were significantly reduced [ (1.04±0.10) vs (0.68±0.06), (0.79±0.09) vs (0.34±0.05), (0.74±0.12) vs (0.48±0.06), P<0.001]; compared with the high glucose+miR-30c group, the cell proliferation activity was significantly reduced in the high glucose+miR-30c+pcDNA3.1-Wnt1 group, and the apoptosis rate was significantly increased [ (0.66±0.07) vs (0.31±0.05), (4.26±0.57) % vs (9.75±0.85) %, P<0.001]. Conclusion:miR-30c may negatively regulate the Wnt/β-catenin signaling pathway, promote cell proliferation, and inhibit cell apoptosis induced by high glucose.

Research progress on the role of miR-30 family in the genesis and development of gastric cancer / 中国肿瘤生物治疗杂志

@#[摘 要] 胃癌（gastric carcinoma，GC）是最常见的消化系统恶性肿瘤之一，其病因及发病机制等都尚未明了，诊断、治疗及预后仍然面临着严峻的问题。微小RNA（microRNA，miRNA）作为一类重要的基因表达调节剂，能够与其相对应的靶基因相结合，从而影响基因表达，以此在GC发生发展中发挥重要的作用。miR-30家族中5个高度保守且成熟的成员——miR-30a、miR-30b、miR-30c、miR-30d和miR-30e位于不同的染色体或邻近位点，这些miRNA在5'末端附近有一个共同序列，3'末端附近具有不同的补偿序列。通过靶向各自相对应的基因影响着GC细胞的增殖、转移、侵袭和对化学药物的耐药性。miR-30家族在GC中发挥着重要的抑癌作用。本文就近年来miR-30家族中5个成员在GC发生发展中作用的研究进展作一综述。

Effects of miR-30c over-expression on early cardiac development of zebrafish / 中华实用儿科临床杂志

Objective To explore the effects of miR -30c over -expression on early cardiac development of zebrafish.Methods The single -cell -stage zebrafish embryos were micro -injected with 3 different concentrations of miR -30c mimics in order to model the miR -30c over -expressed animal.Besides,the effects of miR -30c overex-pression on cardiac development were investigated.The mortality rate and the heart rate were assessed.Beside,the gross morphology and heart morphology of the zebrafish were observed.Moreover,the molecular mechanisms underlying miR -30c function in zebrafish cardiac development was explored,and in -situ hybridization was performed.Results Compared with the wild -type embryos,the mortality rate of zebrafish was increased with the rising miR -30c concen-tration.Furthermore,when the miR -30c mimic concentration was up to 8 μmol/L,the mortality rate reached 80% at 96 h post -fertilization.Simultaneously,the heart rate,which was viewed as an important index to cardiac function,was decreased.Moreover,the pericardial edema was getting worse over time with increasing concentration of overexpression. Then,the cardiac specific gene expression on the zebrafish embryos by whole -mount in situ hybridization was ex-plored.The area of the cardiac chamber was extended and the heart tube looping was negatively affected.Besides,the expression of atrioventricular canal marker genes was absent in zebrafish embryos from miR -30c over -expression groups.Conclusion miR -30c can impact the early cardiac development of zebrafish.

Involvement of miR-30c in resistance to doxorubicin by regulating YWHAZ in breast cancer cells

MicroRNAs (miRNAs) are small RNA molecules that modulate gene expression implicated in cancer, which play crucial roles in diverse biological processes, such as development, differentiation, apoptosis, and proliferation. The aim of this study was to investigate whether miR-30c mediated the resistance of breast cancer cells to the chemotherapeutic agent doxorubicin (ADR) by targeting tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ). miR-30c was downregulated in the doxorubicin-resistant human breast cancer cell lines MCF-7/ADR and MDA-MB-231/ADR compared with their parental MCF-7 and MDA-MB-231 cell lines, respectively. Furthermore, we observed that transfection of an miR-30c mimic significantly suppressed the ability of MCF-7/ADR to resist doxorubicin. Moreover, the anti-apoptotic gene YWHAZ was confirmed as a target of miR-30c by luciferase reporter assay, and further studies indicated that the mechanism for miR-30c on the sensitivity of breast cancer cells involved YWHAZ and its downstream p38 mitogen-activated protein kinase (p38MAPK) pathway. Together, our findings provided evidence that miR-30c was one of the important miRNAs in doxorubicin resistance by regulating YWHAZ in the breast cancer cell line MCF-7/ADR.

The Significance of MicroRNA Let-7b, miR-30c, and miR-200c Expression in Breast Cancers

BACKGROUND: MicroRNA (miRNA) is a class of noncoding protein RNA as a promising biomarker for various diseases. In this study, the expression of let-7b, miR-30c, and miR-200c was studied in breast cancer tissues to evaluate the potential relationship with known clinicopathological parameters. METHODS: Quantitative real-time polymerase chain reaction was performed to determine the expression level of three miRNAs in 37 pairs of noncancerous normal and cancer tissues and an additional 38 cancer tissues from patients with invasive ductal carcinoma. RESULTS: miR-200c expression was higher in cancer tissues compared to noncancerous normal tissues, and its ratio was correlated with patient age at surgery, type of surgery, and Ki-67 expression. The expression level of let-7b in cancer tissues was inversely correlated with lymph node metastasis, histological grade, and Ki-67 expression but positively correlated with estrogen and progesterone receptor expression. miR-200c expression level was positively correlated with Her-2 expression. The miR-30c expression level in breast cancer was not correlated with any parameters. CONCLUSIONS: miR-200c and let-7b could be used as biomarkers in patients with breast cancer, but its pathological mechanism should be determined.