AAV9-mediated erythropoietin gene delivery into the brain protects nigral dopaminergic neurons in a rat model of Parkinson's disease.

We have recently shown that intrastriatal injection of recombinant human erythropoietin (EPO) protects dopaminergic (DA) neurons in the substantia nigra (SN) from 6-hydroxydopamine (6-OHDA) toxicity in a rat model of Parkinson's disease. However, systemic administration of EPO did not protect nigral DA neurons, suggesting that the blood-brain barrier limits the passage of EPO protein into the brain. In the present study, we used an adeno-associated viral (AAV) serotype 9 (AAV9) vector to deliver the human EPO gene into the brain of 6-OHDA-lesioned rats. We observed that expression of the human EPO gene was robust and stable in the striatum and the SN for up to 10 weeks. EPO-immunoreactive (IR) cells were widespread throughout the injected striatum, and EPO-IR neurons and fibers were also found in the ipsilateral SN. Enzyme-linked immunosorbent assay and western blot analyses exhibited dramatic levels of EPO protein in the injected striatum. As a result, nigral DA neurons were protected against 6-OHDA-induced toxicity. Amphetamine-induced rotational asymmetry and spontaneous forelimb use asymmetry were both attenuated. Interestingly, we also observed that intrastriatal injection of AAV9-EPO vectors led to increased numbers of red blood cells in peripheral blood. This highlights the importance of using an inducible gene delivery system for EPO gene delivery.

The increase of miR-27a affects the role of cisplatin on proliferation and migration capacities of liver cancer cells.

OBJECTIVE: To study the effects of chronic virus-mediated micro ribonucleic acid miR-27a on proliferation and migration capacities of liver cancer cells. PATIENTS AND METHODS: A total of 60 patients with primary liver cancer from January 2015 to December 2016 were selected as observation group, 60 patients with chronic liver disease were selected as control group, and another 60 healthy subjects who received physical examination during the same period were selected as healthy group. All patients received serum miRNA detection. The different expressions of serum miRNAs in healthy people, patients with chronic liver disease, and patients with liver cancer were analyzed. The correlations of miR-27a with growth and proliferation of liver cancer MCC-7721 cells were studied. RESULTS: The levels of miR-27a and miR-664b in subjects in control group and healthy group were significantly lower than those in observation group, but the expression levels of miR-30a were statistically elevated (p<0.05). The expression of serum miR-27a in liver cancer patients with higher tumor-node-metastasis (TNM) staging (stage III-IV, number of tumors >1, tumor size >5 cm, and vascular invasion) was significantly higher than those in patients with lower TNM staging (stage I-II, number of tumors =1, tumor size ≤5 cm, and no vascular invasion) (p<0.05). At 48 h and 72 h after transfection, the proliferation of liver cancer MCC-7721 cells was significantly enhanced compared to that in non-transfection group (p<0.05). The level of miR-27a was significantly upregulated in cisplatin-resistant liver cancer A549/CDDP cells compared with that in parental A549 cells. MiR-27a regulated epithelial-mesenchymal transition (EMT) and cisplatin resistance in vitro, while modulated the in vivo response of liver cancer cells to cisplatin. Further studies identified the Raf kinase inhibitor protein (RKIP) as a direct and functional target of miR-27a. The knockdown of RKIP by RNAi showed a similar effect to ectopic miR-27a expression, whereas the over-expression of RKIP weakened the function of miR-27a in liver cancer cells. CONCLUSIONS: MiR-27a participated in the proliferation and migration capacities of liver cancer cells and influenced the effect of cisplatin via targeting RKIP. The high expression of miR-27a is closely related to the malignant degree of liver cancer, which provides guidance for the diagnosis, targeted therapy, and prognostic evaluation of liver cancer.

Nestin expression in different tumours and its relevance to malignant grade.

BACKGROUND: Nestin, an intermediate filament (IF) protein, is expressed in proliferating progenitor cells of developmental and regenerating tissues, and is identified as a neuroepithelial precursor cell marker. Recently, nestin was detected in some neoplasms such as glioma, ependymoma, melanoma, rhabdomyosarcoma, gastrointestinal stromal tumour (GIST), and testicular stromal tumour. Moreover, the expression intensity of nestin exhibited significant correlation with the malignant grade of glioma. AIMS: To detect the expression of nestin in different tumours and to analyse the relationship between the expression of nestin and the malignant grade of the tumours. METHODS: Formalin-fixed and paraffin-embedded surgical samples of neoplastic tissues were obtained from the Department of Pathology of Sun Yat-sen University. Histological analysis and immunohistochemical staining for nestin were performed. Histoscores were analysed by semi-quantitative evaluation. RESULTS: Nestin was expressed predominantly in the cytoplasm of angiosarcoma, pancreatic adenocarcinoma and GIST samples, and some tumour cells expressed in the nucleus. There was a statistically significant difference between the histoscore of nestin in high malignant GIST (2.2366 (0.6920)) and that in low malignant GIST (1.3783 (0.4268)) (p = 0.003); and also between that in high malignant angiosarcoma (1.9188 (0.2069)) and that in low malignant angiosarcoma (0.6474 (0.3273)) (p = 0.000). Cavernous angioma did not express nestin. The histoscore of nestin in high malignant pancreatic adenocarcinoma (7/14) was 1.1767 (0.4676), and that in low malignant pancreatic adenocarcinoma (3/8) was 0.6577 (0.0056) (no significant difference, p = 0.112). CONCLUSIONS: Results suggest that the expression of nestin may play an important role in the development of some neoplasms such as GIST and angiosarcoma.