Inhibitation of effective bcl-2 siRNA on apoptosis of human leukemia-60 cells / 白血病·淋巴瘤

Objective To study the effect of bel-2 siRNA on apoptosis of HL-60 cells.Methods bcl-2 siRNA was synthesized in vitro transcription with silencer siRNA construction kit.The synthesized siRNA was transfected into HL-60 cells with Amine siPORT transfection.We used MTT flow cytometer and hoechst 33258 flourescence stainning t0 evaluate cell proliferation and apoptosis. Results.Bcl-2 siRNA could partially inhibit the growth of HL-60 cells.After incubated with bcl-2 siRNAl for 48 hours,HL-60 cells exhibited morphologic characteristic of apoptosis including chromatin condensation,crescents formation and nuclear fragmentation.Conclusion Effective bcl-2 siRNA can induce apoptosis and inhibit cell proliferation.

Inhibition of bcl-2 gene expression by siRNA in HL-60 cells / 中国药理学通报

Aim To study the interference role of Bcl-2 siRNA on HL-60 cells. Methods Bcl-2 siRNA was synthesized in vitro transcription with silencer siRNA construction kit. The synthesized siRNA was transfected into HL-60 cells with lipid siPORT transfection. Forty-eight hours after the transfection, we used MTT and immunofluorescence to detect cell proliferation and apoptosis,and used RT-PCR and immunofluorescence to detect the level of Bcl-2 mRNA and Bcl-2 protein expression. Result Bcl-2 siRNA reduced the level of Bcl-2 mRNA and Bcl-2 protein expression in HL-60 cells and induced cell apoptosis. There was no difference on the effect of other groups compared with the control. Effective Bcl-2 siRNA specifically degraded Bcl-2 expression in the levels of mRNA and protein and induced HL-60 cells apoptosis.Conclusion These results indicate that siRNA is a highly specific tool for targeted gene knockdown. siRNA-mediated gene silencing is a reliable approach for large-scale screening of gene function and drug target validation.

Change of CD36 expression in minipigs fed high fat/high cholesterol diet / 中国病理生理杂志

AIM: In order to investigate the change of CD36 expression in atherosclerosis. METHODS: Chinese minipigs were fed a normal control diet (CD) or a high fat/high cholesterol diet (HFHC) for 12 months after common carotid artery injury induced by balloon denudation. Plasma total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and triglycerides (TG) were determined by commercially enzymatic methods. CD36 mRNA and protein levels were determined by reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry, respectively. RESULTS: After HFHC for 12 months, plasma total cholesterol, HDL cholesterol and triglyceride in HFHC minipigs were increased compared with the control. CD36 expression and aorta PPAR? in HFHC minipigs were upregulated. CONCLUSION: HFHC may induce hyper cholesterolemia, hypertriglyceridemia and upregulation of CD36 and aortic PPAR? expression.