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Inhibition of PI3K/AKT Pathway Enhanced Through Oxaliplatin Colon Cancer RKO Cell Apoptosis Induced by TRAIL Sensitivity Mechanism

Author: XuMingZuo
Tutor: ZhuZhiTu
School: Liaoning Medical
Course: Oncology
Keywords: TRAIL oxaliplatin colorectal cancer PI3K/Akt apoptosis
CLC: R735.35
Type: Master's thesis
Year: 2013
Downloads: 4
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Abstract


ObjectiveResearch oxaliplatin to TRAIL-induced apoptosis of colon cancer RKO cells, and explore the mechanism of action of the combination, caspase-3protein expression, as well as the impact of the PI3K/Akt pathway.Methods1、MTT assay TRAIL and oxaliplatin inhibit the proliferation of cells.2、Wright-Giemsa staining morphological changes were observed. Flow cytometry to detect apoptosis.3、Western blot method to detect Akt, p-Akt, caspase-3protein expression.4、Statistical analysis:Each experiment was repeated three times, the data to±s, t-test and one-way ANOVA analysis SPSS16.0software.Results1、The inhibitory effect of different concentrations of TRAIL and oxaliplatin single-agent and combination therapy of colon cancer RKO cells:TRAIL role in colon cancer RKO cells, caused only mild apoptosis MTT analysis showed0,25,50,100,200400ng/ml TRAIL role in colon cancer RKO cells for24h, cell proliferation rate were101.9±7.3%,108.8±6.4%,113.2±12.1%,101.5± 5.8%,76.5±5%,73.3±7.2%, and the control group(P>0.05). The results showed that colon cancer RKO cells to TRAIL-insensitive.0,10,20,40,80μg/mL oxaliplatin platinum treatment colon cancer RKO cells, the IC50,24,48and72h, respectively,44.3±7.6,22.3±6.1,9.8±4.5ng/mL, with the control group than P<0.05, the results table Mingaoshali platinum can inhibit the proliferation of colon cancer RKO cells, and was a time-a dose-dependent relationship.100ng/mITRAIL joint40μg/mL oxaliplatin cell proliferation rate oxaliplatin monotherapy group48.4±5.2%to30.6±2.7%, the monotherapy group P <0.05. The results table Mingaoshali platinum enhanced sensitivity to TRAIL-induced colon cancer RKO cell apoptosis.2、Oxaliplatin can cause colon cancer RKO cell apoptosis:Wright-Giemsa staining,40μg/mL oxaliplatin role in colon cancer RKO cells24h can be observed typical apoptotic morphology of apoptotic colon cancer RKO cells, the results table Mingaoshali platinum can induced apoptosis.3、Oxaliplatin enhanced TRAIL-induced colon cancer RKO cell apoptosis:100ng/ml of TRAIL role in colon cancer RKO cell apoptosis ratio by0.9±2.7%only increased to2.5±2.1%, and the control group, P> the0.05,1h,3h,6h,12h,24h, p-Akt activation gradually increased and reached a peak at6hours and12hours,24h began to weaken, there is no observed activation of caspase-3. Tip:TRAIL is involved in colon cancer RKO cells PI3K/AKT pathway, leading to colon cancer RKO cells sensitive to TRAIL.40μg/ml oxaliplatin role in colon cancer RKO cells24h, the apoptosis ratio was9.6±1.9%, with the control group,P<0.05, significantly inhibited the levels of p-Akt can be observed activation of Caspase-3.100ng/ml TRAIL combined with40μg/ml role of oxaliplatin24h, apoptosis ratio increased to23.5±3.5%with monotherapy group P<0.05, also inhibits p-Akt levels, and at the same time can be observed to activation The Caspase-3, and Biaoshali platinum monotherapy group slightly increased. Tip:oxaliplatin can cause colon cancer RKO cell apoptosis, and increase colon cancer RKO cell sensitivity to TRAIL by inhibiting the activation of Akt.ConclusionOxaliplatin by inhibiting TRAIL-induced activation of the PI3K/Akt pathway, enhanced sensitivity of colon cancer RKO cells to TRAIL.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Intestinal neoplasms > Colon tumor
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