| Part I Role of endoplasmic reticulum stress in cell apoptosis induced by Different concentrations of Proteasome inhibitor BortezomibObjective To investigate the role of endoplasmic reticulum stress (ERS) in cell apoptosis induced by Proteasome inhibitor Bortezomib.Methods Culture human glioma cell by way of passage,select glioma cells were treated with Bortezomib._Before the treatment(control group) andl,2,5 and 10nmol/L after the treatment the cellular viability was assessed by MTT assay, PCR and Western-blot was employed to detect ERS associated proteins(GRP78) and apoptosis associated proteins(CHOP). The change of morphology of glioma cells was observed by phase contra_microscope.Result It was revealed by MTT assay that 24h after treatment of Bortezomib, the cell viability of SHG-44 was decreased by 56.2%(P<0.05),and continued to be significantly decreased with concentration(P<0.05).The cell vitality of 5nmol/L group and 10nmol/L group decreased significantly (P<0.05),but between the two group have no statistically significant (P>0.05). The apoptosis rate of 5nmol/L group and 10 nmol/L group decreased significantly (P<0.05),but between the two group have no statistically significant (P>0.05).It was indicated by RT-PCR that ERS associated gene GRP78 and CHOP of SHG-44 cells were significantly increased after treatment of Bortezomib, and continued to be significantly increased with concentration(P<0.05).Western-blot is the same to the PCR.Conclusion ERS may be involved in the apoptosis induced by proteasome inhibitor Bortezomib.5nmol/L is the best role concentrations of Bortezomib.Part II Proteasome inhibitor Bortezomib through CHOP signal transduction pathway induced the endoplasmic reticulum stress-related mechanism analysis in SHG-44 cellObjective To investigate the role of endoplasmic reticulum stress (ERS) in cell apoptosis induced by Proteasome inhibitor Bortezomib. At the same time choose CHOP signal pathway, using the specific CHOP inhibitor Doxorubicin observed changes in the role of Bortezomib, Further analysis of Bortezomib inducing glioma cells apoptosis in the role of the endoplasmic reticulum stress.Methods Culture hunman glioma by way of passage, Experimental group was divided into five groups:normal control group, DTT group, Bortezomib group, Doxorubicin groups, Bortezomib + Doxorubicin Group.after the treatment the cellular viability was assessed by MTT assay, flow cytometry was was employed to detect apoptosis situation.PCR and Western-blot was employed to detect ERS associated proteins(GRP78) and apoptosis associated proteins(CHOP).Results:MTT colorimetric assay was found, 5nmol/L to achieve the best inhibitory concentrations, SHG-44 cell viability decreased 56.2%(P<0.05),After the experimental group, compared with the control group, Bortezomib after 24h role, the apoptotic rate significantly increased (54.35±3.15% vs 3.2±0.42%, p<0.05), GRP78mRNA and CHOP mRNA expression increased significantly (p<0.05); But for DTT, the apoptosis rate and expression of GRP78mRNA and CHOP mRNA has increased (P<0.05). Compared with the DTT group, Doxorubicin + Bortezomib group has been lowered, but not as a separate group of Doxorubicin reduced significantly (p<0.05). Western-blot results showed that compared with the control group, Bortezomib can cause ERS. The role of endoplasmic reticulum stress indicators of GRP78, CHOP expression was significantly increased in the Bortezomib group after 24h (p <0.05).In the DTT group, GRP78, CHOP expression was significantly increased (p<0.05),In the Doxorubicin + Bortezomib group,GRP78, CHOP expression has been reduced, but not as good as giving Doxorubicin group (p<0.05).Conclusions:1. Bortezomib can induced apoptosis of glioma cells.2. Bortezomib can induced apoptosis of glioma cells in the process, ERS is to play the major role of pathways.3. CHOP inhibitor Doxorubicin can partially block the channels of endoplasmic reticulum stress, namely, CHOP signaling pathway is one of the ways to play a role in apoptosis.
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