| Objective: Ovarian cancer is one of malignant tumor fromgynec-ological cancers. In our country, the inception rate of ovarian cancerlocated ter-position from gynecological cancers. At present, the recognitionand diagnosing therapy method on ovarian cancer have gained fairly largeadvancement, but the super-inception rate have no advancement. Tumorcytoreductive surgery after neoadjuvant chemotherapy is the main method oftreatment of ovarian cancer, but because of chemotherapy multidrug resistance(MDR), causing subsequent chemotherapy is failure. Therefore, the researchon the mechanisms of tumor resistance and the reversal of drug resistancebecome a hotspot and difficulty on the study in recent years. Some studyfound that heatshockProtein90(HSP90) as a molecular chaperone plays animportant role in the occurrence and development of tumor in vivo.Geldanamycin (GA) is a specific inhibitor of the HSP90, can block tumorsurvival signal pathways in vivo and have effect of reversal of multidrugresistance. This study is investigated the the influence and mechanism ofreversing drug resistance of geldanamyein combination with cisplatin onhuman ovarian Cancer Cells line SKOV3and SKOV3/DDP, offeringreference basis for the mechanism of drug resistance on ovarian cancer.Methods:1The inhibition effect of proliferation was tested by MTT in differentconcentrations of cisplatin (3,6,9μg/mL), geldanamycin (20,40,200,400,2000nmol/L) and geldanamycin combination with cisplatin on humanovarianCancer Cells line SKOV3and cisplatin resistance on ovarian cancer cell lineSKOV3/DDP, and the reversing multiple RF of geldanamycin combinationwith cisplatin was calculated, RF=IC50(DDP)/IC50(GA+DDP). 2RT-PCR was used to analyze the expression LRP and GST-π mRNA inSKOV3and SKOV3/DDP cells treated by cisplatin, geldanamycin andgeldanamycin combination with cisplatin.3Western-blot was used to analyze the expression LRP and GST-πprotein in SKOV3and SKOV3/DDP cells treated by cisplatin, geldanamycinand geldanamycin combination with cisplatin.Results:1Geldanamycin (20,40,200,400,2000nmol/L) has signifant inhibitioneffects on proliferation of SKOV3, SKOV3/DDP cells. With the increase ofGA concentration, the inhibition effects on proliferation of SKOV3,SKOV3/DDP cells increases gradually on dose depending manner, butdifferent concentrations of geldanamycin have no signifant inhibition effectson proliferation of SKOV3, SKOV3/DDP cells. Different concentrations ofcisplatin (3,6,9μg/mL) have signifant inhibition effects on proliferation ofSKOV3,SKOV3/DDP cells that were treated48hours on dose dependingmanner, and the inhibition effect on SKOV3is more obvious compared withthat on SKOV3/DDP. The reversing multiple RF of geldanamycincombination with cisplatin were2.24,5.52. The results showed thatgeldanamycin combination with cisplatin also have obvious inhibition effectson proliferation of SKOV3, SKOV3/DDP, the inhibition effects obviouslyhigher than single drug of cisplatin and geldanamycin, and can reverse drugresistance of DDP.2Detection of RT-PCR gene level, LRP, GST-π mRNA in SKOV3andSKOV3/DDP cells changes, the results showed that after SKOV3cell treatedwith48h by geldanamycin, cisplatin and geldanamycin combination withcisplatin, the expression of LRP, GST-π mRNA degraded with significantdifference compared with control group (p<0.05). And geldanamycincombined with cisplatin group reducing two resistance genes of LRP, GST-πmRNA was obviously higher than that of single drug of geldanamycin andcisplatin groups; After SKOV3cell treated with48h by above-mentioneddrugs, the expression of LRP, GST-π mRNA degraded with significant difference compared with control group (p<0.05) in geldanamycin andgeldanamycin combination with cisplatin groups. And geldanamycincombined with cisplatin group reducing LRP, GST-π mRNA two resistancegenes was significantly better than that of single drug of geldanamycin group.But the expression of LRP, GST-π mRNA have no obvious decrease incisplatin group with no significant difference compared with control group(p>0.05).3Detection of Western-blot gene level, LRP, GST-π protein in SKOV3and SKOV3/DDP cells changes, the results showed that after SKOV3celltreated with48h by geldanamycin, cisplatin and geldanamycin combinationwith cisplatin, the expression of LRP, GST-π protein degrade with significantdifference compared with control group (p<0.05). And geldanamycincombined with cisplatin group reducing two resistance genes of LRP, GST-πprotein was obviously higher than that of single drug of geldanamycin andcisplatin groups. After SKOV3/DDP cell treated with48h byabove-mentioned drugs, in geldanamycin and geldanamycin combination withcisplatin groups, the expression of LRP, GST-π protein degrade withsignificant difference compared with control group (p<0.05). Andgeldanamycin combined with cisplatin group reducing two resistance genesLRP, GST-π was significantly higher than that of single drug of geldanamycingroup. But the expression of LRP, GST-π protein have no obvious decrease incisplatin group with no significant difference compared with control group(p>0.05).Conclusions:1Geldanamycin combined with cisplatin has signifant inhibition effectson proliferation of SKOV3, SKOV3/DDP cells, the inhibition effects onproliferation of SKOV3, SKOV3/DDP was obviously better than that of singledrug of geldanamycin and cisplatin, and can signifantly reverse drugresistance of DDP.2LRP, GST-mRNA and protein were expressed in SKOV3andSKOV3/DDP cells, but the expression of LRP, GST-π mRNA and protein was striver in SKOV3/DDP cell.3Geldanamycin combined with cisplatin group can signifantly reduce theexpression of resistance genes of LRP and GST-π mRNA in SKOV3andSKOV3/DDP cells, and in the two drugs in the joint group resistance genesLRP, GST-π mRNA and protein expression are stronger than the cut singledrug group.4Geldanamycin can cut through the resistance genes LRP, GST-π ofovarian cancer cells SKOV3/DDP cells with cisplatin resistance and improveovarian cancer cell to cisplatin resistance. |
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