The -160C>a polymorphism in E-cadherin is associated with the risk of nephrolithiasis.

Nephrolithiasis is a common disorder worldwide. E-cadherin (CDH1) is involved in epithelial cell-cell interactions and plays important roles in the etiology of nephrolithiasis. We hypothesized that variants in the CDH1 gene are associated with risk of nephrolithiasis. In a hospital-based case-control study of 127 nephrolithiasis patients and 152 controls frequency matched by age and sex, we genotyped the functional -160C>A (rs16260) polymorphism and assessed its associations with risk of nephrolithiasis in a Chinese population. We found that the CA/AA genotypes were associated with a significantly decreased risk of nephrolithiasis (OR = 0.53, 95%CI = 0.32-0.87), compared with the CC genotype, particularly among subgroups of BMI > 24 kg/m(2) (OR = 0.38, 95%CI = 0.17-0.85), age ≤ 57 years (OR = 0.47, 95%CI = 0.24-0.93), and men (OR = 0.56, 95%CI = 0.29-0.99). Our results suggest that the CDH1 polymorphism is involved in the etiology of nephrolithiasis and thus may be a marker for genetic susceptibility to nephrolithiasis

Mesenchymal Stromal Cells Derived Extracellular Vesicles Ameliorate Acute Renal Ischemia Reperfusion Injury by Inhibition of Mitochondrial Fission through miR-30

Background. The immoderation of mitochondrial fission is one of the main contributors in ischemia reperfusion injury (IRI) and mesenchymal stromal cells (MSCs) derived extracellular vesicles have been regarded as a potential therapy method. Here, we hypothesized that extracellular vesicles (EVs) derived from human Wharton Jelly mesenchymal stromal cells (hWJMSCs) ameliorate acute renal IRI by inhibiting mitochondrial fission through miR-30b/c/d. Methods. EVs isolated from the condition medium of MCS were injected intravenously in rats immediately after monolateral nephrectomy and renal pedicle occlusion for 45 minutes. Animals were sacrificed at 24 h after reperfusion and samples were collected. MitoTracker Red staining was used to see the morphology of the mitochondria. The expression of DRP1 was measured by western blot. miR-30 in EVs and rat tubular epithelial cells was assessed by qRT-PCR. Apoptosis pathway was identified by immunostaining. Results. We found that the expression of miR-30 in injured kidney tissues was declined and mitochondrial dynamics turned to fission. But they were both restored in EVs group in parallel with reduced cell apoptosis. What is more, when the miR-30 antagomirs were used to reduce the miRNA levels, all the related effects of EVs reduced remarkably. Conclusion. A single administration of hWJMSC-EVs could protect the kidney from IRI by inhibition of mitochondrial fission via miR-30

Stratified analyses on association between the <i>CDH1</i>-160C>A polymorphism and risk of nephrolithiasis.

<p>^a Adjusted for age and sex.</p

Downregulation of GAS5 promotes bladder cancer cell proliferation, partly by regulating CDK6.

Long non-coding RNAs (lncRNAs) play important roles in diverse biological processes, such as transcriptional regulation, cell growth and tumorigenesis. However, little is known about whether lncRNA-GAS5 (growth arrest-specific 5) regulates bladder cancer progression. In the present study, we found that the GAS5 expression is commonly downregulated in bladder cancer cell lines and human specimens. Knockdown of GAS5 promotes bladder cancer cell proliferation, whereas forced expression of GAS5 suppresses cell proliferation. We further demonstrated that knockdown of GAS5 increases CDK6 mRNA and protein levels in bladder cancer cells. Expectedly, GAS5 inhibition induces a significant decrease in G0/G1 phase and an obvious increase in S phase. Gain-of-function and loss-of-function studies showed that GAS5 inhibits bladder cancer cell proliferation, at least in part, by regulating CDK6 expression.Downregulated GAS5 promotes bladder cancer cell proliferation, partly by regulating CDK6, and thus may be helpful in the development of effective treatment strategies against bladder cancer

Mass spectrometry analysis of the proteins pull-down by GAS5.

<p>Mass spectrometry analysis of the proteins pull-down by GAS5.</p

The characteristics of patients with bladder cancer.

<p>The characteristics of patients with bladder cancer.</p

GAS5 inhibits bladder tumor cell proliferation.

<p>(A) RT4 cells were treated with GAS5-siRNA, and GAS5 expression level was assayed after 48 h by real-time PCR. (B) RT4 cells were treated with GAS5-siRNA, and at the indicated time points, the numbers of cells per well were measured by the absorbance (450 nm) of reduced WST-8. (C) RT4 cells were treated with pcDNA-GAS5, and GAS5 expression level was assayed by real-time PCR. (D) RT4 cells were transiently overexpressed with GAS5, and the numbers of cells per well were measured by the absorbance (450 nm) of reduced WST-8. The results show data from at least three independent experiments, expressed as the mean ± SD. *<i>P</i><<i>0.05</i>.</p

GAS5 regulates bladder cancer cell cycle by regulating CDK6.

<p>(A) GAS5 expression was inhibited by specific siRNAs in RT4, and the cell apoptosis was analyzed by flow cytometer 48 h later. (B) RT4 cells were treated with GAS5-siRNA or CDK6-siRNA. Forty-eight hours later, the relative cell numbers in each cell cycle phase after propidium iodide staining were determined by FACS analysis. The data are from one of three independent experiments. The histograms were analyzed and the percentage of cells in each phase of the cell cycle is shown. The results are presented as mean ± SD for three experiments.</p

GAS5 negatively regulates CDK6 expression.

<p>(A) Biotinylated GAS5 or control were incubated with nuclear extracts (RT4 cells), targeted with streptavidin beads and associated proteins were resolved in a gel. Silver staining of the SDS-PAGE gel containing aliquots of samples derived from proteins pulled down by GAS5. (B) RIP experiments were performed using CDK6 antibody to immunoprecipitate RNA and a primer to detect GAS5 RNA. (C) Analysis of CDK6 mRNA level was performed in bladder cancer cell lines after GAS5-siRNA treatment. (D) Western blot analysis of CDK6 protein level was performed in bladder cancer cells treated with GAS5-siRNA. (E) Analysis of CDK6 mRNA level was performed in bladder cancer cell lines after GAS5 overexpression. The results show data from at least three independent experiments, expressed as the mean ± SD. *<i>P</i><<i>0.05</i>. (F) Negative correlation between GAS5 levels and the CDK6 levels in 16 bladder cancer samples (<i>r</i>² = 0.168, <i>p = 0.013</i>).</p