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Ginsenoside Rg3 Inhibits Growth Of Human Endometrial Cancer Cell Lines Ishikawa By Inhibiting NF-kappaB Pathway

Posted on:2012-11-28Degree:MasterType:Thesis
Country:ChinaCandidate:J X MaFull Text:PDF
GTID:2154330335499914Subject:Obstetrics and gynecology
Abstract/Summary:PDF Full Text Request
ObjectiveExplore ginsenoside Rg3 (Rg3, GS-Rg3) restrain NF-кB pathway to induce people endometrial cancer cell line Ishikawa apoptosis to provide experimental material for the treatment of endometrial cancer.Methods1. The growth inhibition effect of GS-Rg3 on Ishikawa cells was examined using MTT assways:The Ishikawa cell were processed by different concentrati- ons of GS-Rg3 for 24 hours, 48 hours, 72 hours, were determined absorbency value calculation,and inhibiting rates, drawing concentration response curves.2. To detect the apoptosis of Ishikawa cells: The Ishikawa cells were treated with GS-Rg3 by flow cytometry, Centrifugal, join V-FITC and PI computer detec- tion.3. Ability of invasion was compared with transwell chambermode: After different density GS-Rg3 treating Ishikawa cells 24, 48 hours, made from singlecelled levitation liquid, Transwell matrigel bedding with small room in the fluctuation between the Chambers, chemo tactic agent next room to join, each room with cell liquid, 37℃5 %CO2 training 24 hours, ethanol fixation, Conventional as crystallization violet stain, Counting the number of wear membrane cells, take the average of aggressive tumor cells on every vision at random.4. The protein expressions of caspase-3 and p65 in Ishikawa cells were detected by western blot. After different density GS-Rg3 treating Ishikawa cells 48 hours, collecting the cells, extracting the total protein. Separating and closing nonprotein transferred-printing combining site, 4℃incubation overnight, join antibodies and FaGuangJi, then developed, fuser, experimental repeat 3 times, And gray scanning, scan software analysis with Quanti.Results1. The growth inhibition effect of GS-Rg3 on the Ishikawa cells was examined using MTT assways:GS-Rg3 markedly inhibited the proliferation of Ishikawa cell in vitro (P<0.05).Its effect was more remarkable associated with its concentration increasing, presenting dosage dependence. In certain concentrat- ion the range is dose and time dependence.2. Flow cytometric art detection displayed: After different density GS-Rg3 treating Ishikawa cells 48 hours, its apoptosis rate increase with the concentrat- ion increasing. The results have statistically significant compared with controls difference (P<0.05).3. Cell aggressive comparison: The invasion cell ratio has no significant differences after GS-Rg3 treating Ishikawa cells24 and 48 hours (P>0.05), and the cell attack ability compared with controls differences have statistically significant (P<0.05).4. After different density GS-Rg3 treating the Ishikawa cells 48 hours, measuring each group gray values then do relatively gray- values statistical. The analysis shows that the Caspase-3 proteins in cells with the increase of drug density express increasing gradually, while the P65 protein expression level declined with the increase of GS-Rg3 density compared with the control group, the difference was significant( P<0.05).Conclusions1. GS-Rg3 markedly inhibited proliferation of Ishikawa cell in vitro. Its effect was more remarkable associated with its concentration in creasing, presenting dosage dependence.2. GS-Rg3 can induce Ishikawa apoptosis in vitro.3. GS-Rg3 can put down P65 protein and raise Caspase-3 protein expression,it could inhibit the Ishikawa cell proliferation and promote cell apoptosis by down-regulating the NF-kB signaling pathway.
Keywords/Search Tags:Ginsenoside Rg3, Endometrial cancer, NF-kB pathway, Apoptosis
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