ABSTRACT
Objective:To analyze the expression of hsa-microRNA-6832-5p (hsa-miR-6832-5p) in bladder tumor tissues and bladder tumor cells, and to explore its interference on the expression of proline-rich protein 11 (PRR11) gene in bladder tumor cells and its effect on cell proliferation and migration.Methods:Real-time quantitative polymerase chain reaction (qPCR) was used to detect the expression level of hsa-miR-6832-5p in bladder tumor tissues, paratumorous tissues, bladder tumor cell lines and normal bladder epithelial cells. Bioinformatics predicts and the dual luciferase reporter gene validates the downstream target gene of hsa-miR-6832-5p. hsa-miR-6832-5p or miR-NC were transfected to the bladder tumor cell lines with the lowest expression level of hsa-miR-6832-5p, respectively, and named as miR-6832-5p group and miR-NC group. qPCR was used to detect the expression levels of hsa-miR-6832-5p and target gene mRNA in the transfected cells. Western blot was used to detect the expression level of target gene protein. Methyl thiazolyl tetrazolium (MTT) assay and transwell assay were used to detect cell proliferation and migration, respectively.Results:The expression of hsa-miR-6832-5p was lower in bladder tumor tissues than in adjacent tissues ( P<0.01). The expression of hsa-miR-6832-5p in bladder tumor cells was lower than that in normal bladder epithelial cells ( P<0.05), and T24 cells had the lowest hsa-miR-6832-5p expression level ( P<0.01). Bioinformatics and dual luciferase reporter genes showed that hsa-miR-6832-5p can directly act on the 3′-untranslated region of the PRR11 gene ( P<0.01). The expression of hsa-miR-6832-5p in the miR-6832-5p group was significantly higher than that in the miR-NC group ( P<0.01). The expression of PRR11 in the miR-6832-5p group was significantly lower than that in the miR-NC group ( P<0.01). Western blot results were consistent with qPCR results. Compared with the miR-NC group, the proliferation of bladder tumor cells was significantly decreased after transfection with hsa-miR-6832-5p ( P<0.05), and the migration ability of cells was significantly decreased ( P<0.01). Conclusions:The expression of hsa-miR-6832-5p was significantly decreased in bladder tumor tissues and cell lines. hsa-miR-6832-5p inhibited the proliferation and migration of bladder tumor cells by down-regulating the expression of PRR11 gene.
ABSTRACT
Objective To explore the role of hydrogen sulfide (H 2S ) in acute liver injury induced by crush-ing hind lim bs of rats. Methods The rats w ere random ly divided into the follow ing groups:control, crush-ing, H 2S donor sodium hydrosulfide (NaHS) + crushing, H 2S inhibitor propargylglycine (PAG ) + crush-ing group. The acute liver injury m odel w as established by crushing the hind lim bs of rats w ith standard w eight. R ats w ere sacrificed at 30 m in and 120 m in after the crush. The activities of serum aspartate am inotransferase (AST) and alanine am inotransferase (ALT) w ere m easured by colorim etric m ethod, and the content of H 2S in plasm a and the contents of m alondialdehyde (MDA), protein carbonyl, glutathione (GSH) in the liver and the activity of H 2S generating enzym e (cystathionine γ-lyase, CSE) w ere deter-m ined by chem ical m ethod. The expression of CSEm R N Ain liver w as detected by R T-PCR . Results For crush injury group, the levels of ASTand ALTin serum , MDAand protein carbonyl in liver in-creased. The levels of GSH, CSE, CSEm R N Ain liver and H 2S in serum decreased. The adm inistration of NaHS before lim bs crush could attenuate the changes of liver injury, but the pre-treatm ent w ith PAG could exacerbate the changes. Conclusion The decrease of H 2S production could involve in m ediating the acute liver injury induced by traum atic stress in rats.
ABSTRACT
Objective To investigate effects of antioxidant stress protein hem e oxygenase-1 (HO-1) on lipopolysaccharide (LPS)-induced endoplasm ic reticulum stress (ERS) of rat hepatocytes. Methods The BRL cells (rat hepatocyte cell line) were cultured. The hepatocytes were treated with LPS, LPS+HO-1 si RNA , HO-1 siRNA and PB S solution, respectively. The cell viability was m easured by trypan blue ex-clusion test. The apoptosis cells were detected by the fluorescent dye Hoechst 33258. E xpressions of GR P78, C HO P, caspase-12 and HO-1 were detected by Western blotting. Results LPS caused an in-crease of HO-1 protein expression of rat hepatocytes in a dose-dependent and tim e-dependent m anner, a up-regulation of GRP78, CHO P and caspase-12, a decrease in cellviability,and an increase in apopto-sis rate of hepatocytes. Pretreatm ent of HO-1 siRNA inhibited the up-regulation of LPS-induced HO-1, however, aggravated ERS and cellular injury. Conclusion HO-1 inhibites ERS-m ediated cellular injury of rat hepatocytes induced by LPS.