MiR-335-5p inhibits proliferation of Huh-7 liver cancer cells via targeting the Oct4/Akt pathway.

OBJECTIVE: The aim of this study was to detect the expression of micro ribonucleic acid (miR)-335-5p in the liver tissues of patients with liver cancer, and to explore its effect on liver cancer and mechanism using Huh7 human liver cancer cells. PATIENTS AND METHODS: Liver tissues were collected from patients with liver cancer. The expression of miR-335-5p in tissues was detected via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Subsequently, Huh7 cells were transfected with miR-335-5p in vitro. After overexpressing miR-335-5p, changes in the expression of octamer-binding transcription factor 4 (Oct4) gene were observed via qRT-PCR. Furthermore, the proliferation of Huh7 cells and the protein expressions of protein kinase B (Akt) and phosphorylated Akt (p-Akt) were detected using cell counting kit (CCK)-8 assay and Western blotting (WB), respectively. RESULTS: Compared with Control group, the expression of miR-335-5p increased significantly in the liver tissues of liver cancer patients (p<0.01). In comparison with those in negative group, the messenger RNA (mRNA) expression of Oct4 and the proliferation rate of Huh7 cells were both significantly inhibited in miR-335-5p group (p<0.01, p<0.05). After overexpression of miR-335-5p, the protein expression level of p-Akt decreased remarkably (p<0.01). CONCLUSIONS: MiR-335-5p directly binds to the 3' untranslated region (3'UTR) of Oct4 mRNA to restrain the phosphorylation of Akt, thereby inhibiting Huh7 cell proliferation.

Phosphorylated AKT inhibits the apoptosis induced by DRAM-mediated mitophagy in hepatocellular carcinoma by preventing the translocation of DRAM to mitochondria.

Increasing autophagy is beneficial for curing hepatocellular carcinoma (HCC). Damage-regulated autophagy modulator (DRAM) was recently reported to induce apoptosis by mediating autophagy. However, the effects of DRAM-mediated autophagy on apoptosis in HCC cells remain unclear. In this study, normal hepatocytes (7702) and HCC cell lines (HepG2, Hep3B and Huh7) were starved for 48 h. Starvation induced apoptosis and autophagy in all cell lines. We determined that starvation also induced DRAM expression and DRAM-mediated autophagy in both normal hepatocytes and HCC cells. However, DRAM-mediated autophagy was involved in apoptosis in normal hepatocytes but not in HCC cells, suggesting that DRAM-mediated autophagy fails to induce apoptosis in hepatoma in response to starvation. Immunoblot and immunofluorescence assays demonstrated that DRAM translocated to mitochondria and induced mitophagy, which led to apoptosis in 7702 cells. In HCC cells, starvation also activated the phosphatidylinositol 3-kinase (PI3K)/AKT pathway, which blocks the translocation of DRAM to mitochondria through the binding of p-AKT to DRAM in the cytoplasm. Inactivation of the PI3K/AKT pathway rescued DRAM translocation to mitochondria; subsequently, mitochondrial DRAM induced apoptosis in HCC cells by mediating mitophagy. Our findings open new avenues for the investigation of the mechanisms of DRAM-mediated autophagy and suggest that promoting DRAM-mediated autophagy together with PI3K/AKT inhibition might be more effective for autophagy-based therapy in hepatoma.

Dietary supplementation with Lactobacillus fermentum I5007 improves the anti-oxidative activity of weanling piglets challenged with diquat.

AIMS: To determine the effects of Lactobacillus fermentum I5007 on the redox state of piglets oxidatively stressed with diquat. METHODS AND RESULTS: Twenty-four, 28-day-old barrows were used in a 2 × 2 factorial design experiment with the main effects being Lact. fermentum supplementation and diquat challenge. Half of the pigs (n = 12) were orally administered with 20 ml of a solution containing 10(8 ) CFU ml(-1) of Lact. fermentum each morning of the 21-day trial, while the remainder received saline. On day 8, these two groups were further subdivided so that half of the pigs in each group (n = 6) were intraperitoneally injected with 10 mg kg(-1) BW diquat, while the remainder received saline. The diquat-injected pigs had significantly poorer performance and increased levels of plasma cortisol, adrenaline, carbonyl and malondialdehyde. Lactobacillus fermentum supplementation significantly increased superoxide dismutase and glutathione and increased the ability to inhibit superoxide anion production in liver and muscle. CONCLUSIONS: Lactobacillus fermentum improved the anti-oxidative defence system and alleviated damage caused by diquat. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus fermentum has the potential to alleviate oxidative stress and improve weaning pig performance.

Upward herniation of the posterior fossa cyst in the shunted child.

Seventy-two years after Dandy and Blackfan's initial description of the Dandy-Walker syndrome, controversies still exist regarding the precise methods of treatment for this disorder. One potential complication of simple lateral ventricular shunting is upward herniation of the posterior fossa cyst, which at times can result in precipitous clinical deterioration. We present three examples of posterior fossa cysts with upward herniation and review the pertinent literature. We advocate contrast studies to determine whether aqueductal communication exists, before shunting is performed, as well as close follow-up of the shunted child.