ABSTRACT
AIM:To investigate the molecular mechanism of inhibition of miR-421 expression promoting radiosensitivity in the cervical cancer cells.METHODS:Cervical cancer lines HeLa, SiHa, C33A and CaSki were transfected with miR-421 inhibitor or negative control nucleotide using Lipofectamine 2000 kit, and the levels of miR-421 expression in the cervical cancer lines and endometrial epithelium cell line ESC were detected by real-time PCR.These cells with transfection were exposed to various doses of X-ray (0, 2, 4, 6 and 8 Gy).After 48 h, the cell viability, LDH leakage rate and apoptotic rate were measured respectively by MTT assay, ELISA and flow cytometry with Annexin V-FITC/PI staining.The protein levels of cleaved caspase-9, caspase-9, cleaved PARP, PARP, Bcl-2 and Bax were monitored by Western blot.RESULTS:Low miR-421 levels was found in the ESC cells, while high miR-421 levels were observed in the HeLa, SiHa, C33A and CaSki cells.The level of miR-421 in the cells transfected with miR-421 inhibitor was significantly lower than that in negative control group (P<0.05).The viability and LDH leakage rate of the cervical cancer cells with low miR-421 expression were notablely lower than those in negative control group, and the apoptotic rate at 72 h was remarkablely increased (P<0.05) under the same conditions.The results of Western blot indicated that, after exposure to ionizing radiation, the protein levels of cleaved caspase-9, cleaved PARP and Bcl-2 were significantly increased, while the protein level of Bax was significantly decreased in the cervical cancer cells with low miR-421 expression compared with negative control group (P<0.05).CONCLUSION:miR-421 is lowly expressed in the normal endometrial epithelial cells, but highly expressed in the cervical cancer cells.Down-regulation of miR-421 expression significantly inhibits the growth and enhances the radiosensitivity of cervical cancer cells at least partly via activating caspase-9 apoptosis pathway, thus promoting Bcl-2 and inhibiting Bax expression.
ABSTRACT
Objective Motility is an important physiological characteristic of a mature sperm.Nerve growth factor(NGF) is a protein essential for the development,maintenance and survival of the peripheral and central nervous systems.NGF and its receptors TrkA and p75 are widely expressed in the testis,accessory reproductive organ,and the epididymal sperms.In the present study,we investigated the role of NGF on human sperm motility.Methods Use 0.1,1 and 10 μmol/L concentrations of NGF,on sperm motility study to investigate the optimal concentration.Use CASA to detect Sperm motility changes every 10 minutes in an hour after 10 μmol/L NGF was added to the semen.Results The parameters of sperm motility increased after NGF incubation had significant difference, in particular,VAP,VSL,VCL,BCF and LIN mean were significantly increased more than 32%.MAD,STR,ALH and WOB mean had no notable difference.Furthermore,NGF promotes the sperm motility in a time- and dose- dependent manner.In addition,the enhancement of NGF on sperm motility was more stronger than those of sperm culture medium.Conclusion Our findings suggest that NGF plays a promoted role in human sperm motility.
ABSTRACT
Atherosclerosis is a pathegenesis process involved in the inflammatory and proliferative responses of cells in which endothelial cells have an important function in the regulation of a variety of genes. A variety of molecules have been identified in the atherosclerotic environment that are able to activate NF ?B. Possible functional implications for activated NF ?B in atherogenesis are discussed by protecting endothelial cells selectively, perhaps, it may provide a new method for the therapy of the atherosclerosis.