Roles of Autophagy and Protein Kinase C-epsilon in Lipid Metabolism of Nonalcoholic Fatty Liver Cell Models.

OBJECTIVE: To investigate interaction between autophagy and PKC-ε in lipid metabolism of NAFLD cell models. METHODS: HL-7702 cells and SK-HEP-1 cells were cultured in vitro as NAFLD cell models and treated with RAPA to induce autophagy. 3-MA was used to inhibit cell autophagy. And HL-7702 and SK-HEP-1 cell were ordinary cultured as control groups. Cell viability was determined by MTT colorimetric assay. The levels of TG, TC and PKC-ε were detected by ELISA. PKC-ε was detected by IF. LC3-II/LC3-I, P62, IRS-1, IRS-2, PI3Kp85, mTOR were detected by Western-blot. SPSS 20 software was used for statistical analysis. RESULTS: The values of PKC-ε were the highest in the steatosis groups (HL-7702 cells were 91.10%, SK-HEP-1 cells were 98.20%). Compared with the steatosis groups, the LC3-II/LC3-I ratio in the induced autophagy groups increased obviously (p <0.05). P62/ß-actin grayscale ratio of the induced autophagy groups decreased significantly compared with the steatosis group (p <0.05). MTOR/ß-actin grayscale ratio in the induced autophagy groups were significantly lower than those in the steatosis groups (p <0.05). PI3Kp85, IRS-1 and IRS-2/ß-actin grayscale ratio of the induced autophagy groups increased significantly compared with the steatosis group (p <0.05). CONCLUSION: Up-regulation of autophagy can promote the elimination of liver fat; while down-regulation can promote lipid accumulation. The expression of PKC-ε is positively related to the degree of hepatic steatosis. PI3K was involved in both autophagy and IR induced by PKC-ε. PKC-ε might participate in hepatocyte autophagy by regulating PI3K.

Overexpression of PI3K-p110α in the progression of uterine cervical neoplasia and its correlation with pAkt and DJ-1.

OBJECTIVE: To investigate the expression of PI3K-p110α, pAkt, PTEN, the signaling molecules from PI3K/Akt signaling pathway, DJ-1, an oncoprotein and HSP90a, a molecular chaperone, and their correlation in uterine cervical neoplasia, in order to elucidate their role in cervical carcinogenesis. MATERIALS AND METHODS: Using immunohistochemistry, the authors analyzed the expression of PI3K-p110α, pAkt, PTEN, DJ-1 and HSP90α, and their correlation in ten normal tissues, cervical intraepithelial neoplasia (CIN) including 30 CIN1 and 31 CIN3, and 33 cases of invasive squamous cell carcinoma (SCC). RESULTS: The expression of all proteins significantly increased in CIN3 compared to CIN1, and only the expression of PI3K-p110α significantly increased in invasive SCC compared to CIN3. There was a significant positive correlation between the expression of PI3K-p110α and DJ-1, as well as PI3K-p110α and pAkt in CIN3 and invasive SCC. CONCLUSION: Overexpression of PI3K-p110α is associated with progression of uterine cervical neoplasia, and the expression of pAkt and DJ-1 is positively correlated with PI3K-p110α expression in this process.

Adenovirus mediated knockdown of bone morphogenetic protein 2 inhibits human lung cancer growth and invasion in vitro and in vivo.

Bone morphogenetic protein 2 (BMP-2) is a member of the TGF-beta superfamily of signaling molecules, and has been shown to function as a tumor suppressor involved in development and progression of many malignancies. BMP-2 has previously been reported to be closely correlated with lung cancer. But, the role and molecular mechanisms of BMP-2 in lung cancer have not yet been comprehensively explained. The present study aims to elucidate the role of BMP-2 in growth and invasion of human lung adenocarcinoma (LAC) in vitro and in vivo. Adenovirus vector-mediated BMP-2 small hairpin RNA (shBMP-2) was used to transfect into A549 LAC cells to determine the functional relevance of BMP-2 and tumor growth and invasion in vitro and in vivo, and further investigate the expression levels of BMP-2, vascular endothelial growth factor (VEGF), matrix metallopeptidase-9 (MMP-9), phosphatidylinositol 3-kinase p85alpha (PI3Kp85alpha) and phosphorylated AKT (p-AKT). As a result, LAC cell proliferation and invasion were significantly diminished by knockdown of BMP-2 indicated by MTT and Transwell assays, and cell apoptosis and cycle arrest were markedly induced indicated by flow cytometry. When BMP-2 expression was knocked down, the expression of PI3Kp85alpha, p-AKT, VEGF and MMP-9 was also down-regulated in LAC cells. In addition, the tumor volumes in LAC subcutaneous nude mouse model treated with shBMP-2 were significantly smaller than those in control and ad-GFP groups. Taken together, our findings indicate that knockdown of BMP-2 inhibits growth and invasion of LAC cells possibly via blockade of the PI3K/AKT signaling pathway, and BMP-2 may be a potential therapeutic target for lung cancer.