ABSTRACT
Objective To investigate the effect of knockdown or overexpression of G6PD on proliferation, growth and migration of human hepatocellular carcinoma cell PLC/PRF/5. Methods Lentivirus-mediated knock-down or overexpression of G6PD was achieved in human hepatocellular carcinoma cell line PLC/PRF/5. RT-PCR and Western blotting assay were used to detect the overexpression or knockdown of G6PD.Cell proliferation and mi-gration curves were recorded by real-time cell analysis system(RTCA),the cell proportion in the DNA replication phase can be directly displayed with EDU experiment,cell growth ability was detected by colony forming assay. Results The doubling time of cells in G6PD knockdown group was longer than that of the control group,and the cell growth rate decreased significantly,the proportion of cells in proliferative phase(43.2%)was lower than that in the control group,but the rates colony formation and migration were significantly decreased(P<0.05,respective-ly),and the migration curves separated apparently.While no significant differences in proliferation,growth and mi-gration of PLC/PRF/5 cells were found between the over-expressed strain and the control group. Conclusion The reduction of G6PD expression in HCC cells inhibits the proliferation and growth of HCC,which may lay a foun-dation for the further study of the pathogenesis and treatment of HCC.
ABSTRACT
Objective To compare and analyze the primary and secondary structures and antigenic epitopes of the two allergens: Der p 2 and Der f 2. Methods The protein sequences of Der p 2 and Der f 2 were downloaded online. The primary and secondary structures of the dust mite allergens were compared and analyzed bioinformatically to determine the potential epitope and signal peptide sites. Results Both Der p 2 and Der f 2 contained 146 amino acids and 9 potential protein binding sites with a secondary structure that mainly contains [3 - sheets, and there might be signal peptides site at the 1st 17th segment of the N - terminus. B cell epitopes analysis revealed that both Der p 2 and Der f 2 have 9 potential linear B epitopes and 2 conformational B epitopes. NetMHCⅡserver prediction showed Der p 2 contains 6 high affinity sites, whereas Der f 2 0nly contains 5. Conclusion This study may lay the foundation for further research of the biochemical function of the 2 allergens and contribute to vaccine development for allergen - specific immunotherapy.
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Objective To explore the potential impact of low concentration of metformin on the morphology and function of mitochondria of HepG2 cells. Methods HepG2 cells in experimental group and control group were treated with or without low concentration of metformin (1mM/L), respectively. The cells were incubated for 12h in the incubator with constant temperature and humidity as well as 1% oxygen. Orange Mitoview was used to stain the mitochondria to detect the effects of the drug on its morphology and quantity. Transmission electron microscope was utilized to observe the effect of metformin on the ultrastructure of mitochondria. The mitochondrial respiratory chain complex I activity in HepG2 cell was detected by Complex I Enzyme Activity Dipstick Assay Kit (DAK). Results Orange Mitoview staining showed that low concentration of metformin had little effect on the morphology and number of mitochondria of cells in experimental group , and the difference between control and experimental group was not statistically significant (P > 0.05). In addition, the result was further determined by transmission electron microscopy. However, DAK analysis showed that complex I activity of cells in experimental group was significantly lower than that in control group. Conclusion Under Hypoxia conditions, low concentration of metformin had no significant effect on the morphology and number of mitochondria of HepG2 cells, but it significantly reduces the activity of mitochondria of HepG2 cells.