Effects Of C086 Combined With Imatinib On GIST-882 Cells

Objective: Gastrointestinal stromal tumor(GIST)is the most common mesenchymal tumor in the digestive tract.Imatinib Mesylate(IM)is a first-line targeted therapy for patients with moderate-and high-risk GIST,which can significantly prolong the median survival and progression-free survival of patients with GIST.However,studies have shown that the incidence of drug resistance in IM targeted therapy is as high as 70%-80%,which may eventually lead to the progression of recurrence of GIST.Currently,these patients with GIST often lack further effective treatment methods.In recent years,studies have found that heat shock protein 90 is expressed in a variety of tumor tissues including GIST,and it has been reported that it is closely related to tumor development,drug tolerance and treatment prognosis.C086 is a curcumin derivative synthesized by Institute of Pharmacy,Fujian Medical University.It has strong HSP90 inhibitor activity.In summary,this study will investigate the role of HSP90 in the development and progression of GIST and targeted drug resistance by observing changes in the GIST-882 sensitive and resistant cell lines of GIST-882 after the specific inhibitor of HSP90,C086.The effects of GIST882 cells provide new ideas and theoretical basis for the treatment of imatinib-resistant GIST.METHODS:1.Paraffin sections were prepared from surgical specimens of 32 patients with GSIT who had been treated for the first time in our hospital,who had not undergone targeted therapy or chemotherapy prior to surgery on 2010.1-12.HSP90 was detected in the tumor tissues and normal tissues by immunohistochemistry.The expression of HSP90 in tumor tissues and normal tissues from patients with GIST was compared;2.The HSP90 inhibitor C086,IM were tested for GIST-882 imatinib resistance and sensitive cell lines by cell counting and CCK-8.The effect of cell proliferation was calculated,and the inhibition rate of C086 and IM on GIST-882 was calculated.Then the growth curve was calculated and IC50 was calculated.The proliferation of imatinib-resistant and sensitive cell lines of GIST882 was observed by C086,IM single drug and combination therapy.Different effects;3.The cell cycle was detected by flow cytometry,and the effects of C086,IM alone and combination drugs on the cell cycle and apoptosis of GIST882 imatinib-resistant and sensitive cell lines were observed;4.Changes of signal pathway molecules in GIST882 imatinib-resistant strain IM-1 cells treated with concentration C086.Results:1.The expression of HSP90 in normal tissues of 32 patients with GIST was significantly lower than that of tumor tissues(P<0.05);2.The proliferation of GIST882 cell line was significantly inhibited by C086,and the inhibitory effect was increased with the concentration of C086.Increasingly,the combination of C086 and Imatinib in GIST-882 imatinib-resistant and sensitive cell lines was more potent than C086 and Imatinib alone(P<0.05).3.Scratch test results showed: C086 There was no significant effect on the lateral migration ability of GIST882 resistant and sensitive cell lines.4.Flow cytometry results showed that there was no significant difference in G0/G1 cell ratio between C086 and Imatinib treatment groups(P>0.05).GIST-882 IM-1 cells in the S phase and G2 phase treated cells were significantly lower than the control group(P<0.05);5,C086 treatment groups HSP90,Akt,P-Akt,P-Erk expression levels were significantly lower(p <0.05),and with the increase of C086 concentration,there was a decreasing trend;the expression level of KIT and P-KIT in C086 treatment group was significantly decreased(p<0.05),but the degree of change did not change with the increase of C086 concentration,but Erk’s The expression level is significant except for C086 100umol/l treatment Lower(p <0.05),under which conditions the expression level of other concentrations did not change significantly(p> 0.05)conclusion:1.The expression of HSP90 in high-risk patients with GIST was significantly higher than that in positive tissues(P<0.05).2.C086 treatment of GIST imatinib-resistant and sensitive cell lines,can make the cell proliferation decreased,while the combination of C086 and Imatinib in the resistant strains inhibited cell proliferation better than C086,Imatinib single drug effect Case;3.HSP90 may promote cell proliferation mainly through Akt-mediated PI3K/Akt and RAF/Erk pathways.