| Objective1.to study the effects of antioxidant reduced glutathione(GSH) on the anti-proliferation and anti-apoptosis role of doxorubicin (DOX) treat thehuman hepatoma cell line HepG2；2.to investigate the gene expression of NF-κB p65、bcl-2after HepG2cell line apoptosis induced separately by both single DOX treatment and itscombination with GSH；furthermore，based on these gene expression change，tofurther explore the mechanism of their interaction.3.to try to provide a case study onthe notion of individualized hepatoprotection during cancer chemotherapy. Methods1.MTT assay was used to probe the HepG2cell growth after being treated by variousconcentrations of DOX and combination with GSH;2.Real-time RT-PCR was used to investigatethe NF-κBp65mRNA expression changes in the HepG2cells after being treated by variousconcentrations of single DOX and combination with GSH100μg/ml；3. Western blotting was usedto document the protein expression changes of NF-κB p65and Bcl-2in the HepG2cells at the24th hour after treated by single DOX2μg/ml dosage plus its combination with GSH100μg/ml；4. Flow cytometry was used to study the apoptosis of HepG2cells at the24th hour after beingtreated by DOX2μg/ml and its combination with GSH100μg/ml. The experimental data wasanalyzed by SPSS version19.0software and both analysis of variance and SNK-q statistic wereused to compare the means between groups. Results The rate of growth inhibition wasmeasured1.MTT method: When (1) DOX concentration0.125μg/ml,0.25μg/ml,0.5μg/ml,1μg/ml,2μg/ml, the role of24h,48h,72h inhibited proliferation of HepG2cells; these different concentrations of DOX, respectively, and GSH (100μg/ml)joint, the role of24h,48h,72h also inhibited the proliferation of HepG2cells, but theinhibition rate of each group compared with DOX alone has reduced. DOX monotherapy group, the inhibition rate DOX joint GSH group has time and dose-dependent manner, and at different time points between the two groups at the sameconcentration of drug inhibition rate was statistically significant (P <0.05); at the sametime two different concentrations there are significant differences between the groupsinhibition rate (P <0.05);(2) DOX monotherapy group was higher than DOXinhibited joint GSH group, the two groups were statistically significant (p <0.05).2Flow cytometry DOX2μg/ml, and DOX2μg/ml joint GSH100ug/ml for24h afterHepG2cells were apoptotic rate. The control group apoptosis was （0.733±0.153）%，DOX monotherapy group apoptosis was （28.400±0.007）%, apoptoticcells in the combination group was （15.500±0.006）%, the two groups werestatistically significant (P <0.01) compared with the control group，DOX monotherapygroup was statistically significant (P <0.01) compared with the DOX joint GSH group.3RT-PCR results showed that, DOX monotherapy group, DOX joint GSH grouptreated HepG2cells after24h, the expression of NF-κB p65mRNA two groups weresignificantly different (P <0.05)4Western blotting results showed that:. DOX2μg/ml,and DOX2μg/ml joint GSH100μg/ml treated HepG2cells after24h, expression ofNF-κB p65: DOX group than in the control group increased expression, DOX+GSHgroup expression increased expression compared with DOX group. The two groupswere statistically significant (P <0.05) compared with the control group， DOXmonotherapy group was statistically significant (P <0.05) compared with the DOXjoint GSH group. Bcl-2expression: expression DOX group than in the control groupincreased, DOX+GSH group expression increased expression compared with DOXgroup. The two groups were statistically significant (P <0.05) compared with thecontrol group， DOX monotherapy group was statistically significant (P <0.05)compared with the DOX joint GSH group. Conclusion The combination of DOX withGSH will reduce its anti-cancer effects via possibly upregulation of NF-κB p65andBcl-2gene expression；Therefore for those patients who was given DOX to treatcancer should avoid using GSH to protect liver at the same time. |
PDF Full Text Request