ABSTRACT
Objective:To explore the effect of miR-1249-5p on the proliferation, metastasis and cell cycle of PC-3 cell in prostate cancer.Methods:The relationship between the expression level of miR-1249-5p and the overall survival of prostate cancer patients was analyzed using OncoMir Cancer Database (OMCD). The human prostate cancer cell line PC-3 was divided into two groups: miR-1249-5p group and negative control group. Mediated by Lipofectamine 2000, miR-1249-5p mimics liposome complex or negative miRNA liposome complex were transfected into PC-3 cell at logarithmic growth stage. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-1249-5p in PC-3 cell of two groups. Colony formation assay was used to detect the changes of the proliferation ability of PC-3 cell in the two groups. Transwell experiment was used to detect the changes of PC-3 cell invasion in the two groups, and the cell cycle changes of the two groups of PC-3 were detected by flow cytometry. The miRNA prediction software miRGator was used to predict the target gene of miR-1249-5p. RT-qPCR and Western blotting were used to detect the target gene expression of miR-1249-5p. Measurement data were expressed as mean±standard deviation ( ± s), and t-test was used for comparison between two groups. Results:Compared with prostate cancer patients with low miR-1249-5p expression, prostate cancer patients with higher miR-1249-5p expression had longer overall survival, and the difference was statistically significant ( P<0.01). The expression level of miR-1249-5p in the miR-1249-5p group (10.74±1.19) was significantly higher than that of the negative control group (1.56±0.27), the difference was statistically significant ( P<0.01). The number of colonies formed in the miR-1249-5p group (35.86±6.94) was significantly less than that in the negative control group (88.94±11.66), and the difference was statistically significant ( P<0.01). The number of transmembrane cells [(25.01±6.83)/high power field of view] in the miR-1249-5p group was significantly less than that of the negative control group [(82.76±8.35)/high power field of view], and the difference was statistically significant ( P<0.01). The proportion of cells in the G 0-G 1 phase in the miR-1249-5p group [(50.79±6.61)%] was significantly higher than that in the negative control group [(27.09±2.30)%], the difference was statistically significant ( P<0.01), and PC-3 cell were inhibited in the G 0-G 1 phase. Neural precursor cell expressed developmentally down-regulated 9 ( NEDD9) may be the target gene of miR-1249-5p. Compared with the negative control group, the NEDD9 gene expression in the miR-1249-5p group was significantly lower than that of the negative control group, the difference was statistically significant ( P<0.01). Conclusion:miR-1249-5p can inhibit the proliferation, metastasis and cell cycle of PC-3 cell in prostate cancer, which may be achieved by negatively regulating the expression of proto-oncogene NEDD9.
ABSTRACT
Neural precursor cell Expressed‚Developmentally Down-regulated protein 4 (NEDD4-1‚ also known as NEDD4 in some papers) is a tumor-related protein that has attracted much attention in recent years. It belongs to the E3 HECT (homologous to E6 associated protein C terminus) ubiquitin ligase‚ which could ubiquitinate various proteins that are subsequently degraded in lysosomes or proteasomes‚ or mediate their nuclear-cytoplasmic translocation‚ or indirectly affect various signaling pathways of different malignant tumors. With the deepening of a large number of tumor-related experiments‚ it has been found that NEDD4-1 can affect the biological behavior of tumors by regulating cell cycle‚ invasion and metastasis of cancer cells‚ antagonize drug resistance and many other pathways. In digestive system tumors‚ NEDD4-1 mainly promotes the proliferation‚ invasion and migration of hepatocellular carcinoma through multiple pathways such as PTEN/ PI3K/ Akt‚ TGF-β‚ Hippo and LDLRAD4. In pancreatic cancer‚ NEDD4-1 acted as an oncogene in the PI3K/ Akt signaling pathway‚ but acted as a tumor suppressor gene in the Myc-Sirt2 signaling circuit. In gastric and colorectal cancer‚ the NEDD4-1-related signaling pathways are different from other digestive system tumors. NEDD4-1 promotes gastric cancer progression and metastasis (via the EGFR signaling pathway) and inhibits colorectal cancer tumor growth (via the Wnt signaling pathway) independently of the PTEN/ PI3K/ Akt pathway. NEDD4-1 has become a hot research direction for therapeutic purposes. In this paper‚ we summarize the functions‚ signaling pathways and potential inhibitors of NEDD4-1 in different digestive system tumors‚ and discuss the relationship between NEDD4-1 and different signaling pathways‚ aiming to provide important reference data for cancer therapy.
ABSTRACT
BACKGROUND: Whole-genome expression profiling is a technical method for gene expression research, with high sensitivity and specificity. This technique can be used to detect differential genes related to chronic periodontitis in the whole genome, therefore efficiently and quickly finding chronic periodontitis-related factors. OBJECTIVE: To screen genes related to chronic periodontitis by using the whole-genome expression profiling. METHODS: Normal periodontal ligament tissue of 15 patients with orthodontic extraction was selected as control group, and periodontal tissue of 21 patients with chronic periodontitis was selected as experimental group. To screen up-regulated and down-regulated genes. the genome-wide expression profile chips of four chronic periodontitis tissues and four healthy tissues were compared. The expression of the differential gene PI3K-Akt signal pathway was verified by real-time PCR (7 normal cases and 13 cases of chronic periodontitis) and western blot (4 normal cases and 4 cases of chronic periodontitis). The experimental protocol was approved by the Ethics Committee of the First Affiliated Hospital of Hainan Medical University (approval No. HNM20180034) and informed consent was obtained from each patient. RESULTS AND CONCLUSION: Analysis of the whole genome expression profile chip revealed that 1 565 up-regulated genes and 1 849 down-regulated genes were significantly differentially expressed in chronic periodontitis samples. The enrichment analysis revealed that the expression of PI3K-Akt signaling pathway was significantly different in chronic periodontitis (P < 0.001). Real-time PCR and western blot assay results indicated that PI3K and Akt expression was higher in the experimental group than in the control group (P < 0.05). All the findings indicate that the genome-wide expression profile chip is fast and highly sensitive to screen the changes in chronic periodontitis-related genes. Significantly differential expression of PI3K-Akt signal pathway in chronic periodontitis provides an experimental basis for the treatment of chronic periodontitis.
ABSTRACT
OBJECTIVE: To evaluate and compare protein expression profiles of synovial fibroblasts using proteome analysis in swine knee injuries with joint instability, during early post-traumatic arthritis (PTA) development. METHOD: Experimental PTA was induced by transection of the anterior cruciate ligament (ACL) in swine left knee joints. After sacrifice at 8 weeks, cartilage and synovium obtained from both knee joints were prepared for histopathologic examination. Cultured synovial fibroblasts were processed for 2-dimensional electrophoresis and mass spectrometric analysis. Histopathologic examination showed overt arthritic changes that supported the development of early PTA. RESULTS: Proteome analyses led to the identification of more than 1,500 protein spots and of 11 differently expressed protein spots. Of those, six proteins were down-regulated (cytoskeletal beta actin, cofilin-1, destrin, Rho GDP dissociation inhibitor alpha, and unnamed protein product), and five proteins were up-regulated (alpha-B crystallin, smooth muscle protein 22-alpha, and cytoskeletal beta actin) in ACL-transected synovial fibroblasts. That is, proteins related to cellular organization and signal transduction are down-regulated, and those related to cell rescue, defence, and stress are up-regulated. CONCLUSION: These results may suggest that joint instability contributes to the development of PTA and is one of the major etiologic factors of PTA. In addition, this suggests that the proteome analysis of synovial fibroblasts is a useful approach in examining a joint after an injury and can be used to understand the pathogenesis of PTA.