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The Effects of Celecoxib on the Proliferation and the Reversing Multidrug Resistance in Drug-resistance Human Ovarian Cancer Cell SKOV-3/DDP

Author: GuoXiangMei
Tutor: LiuFengLing
School: Hebei Medical University
Course: Oncology
Keywords: resistance human ovarian cancer cell line SKOV-3/DDP apoptosis reverse multiple drugresistance Survivin GST-π
CLC: R737.31
Type: Master's thesis
Year: 2011
Downloads: 75
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Abstract


Objective: To study whether the selective COX-2 inhibitor celecoxib can inhibit the proliferation of cisplanin resistance human ovarian cancer cell line SKOV-3/DDP and possible mechanism;to observe the effects of celecoxib on reversing the cisplatin resistance in human ovarian cancer cell line SKOV-3/DDP and possible mechanism.Method:1 SKOV-3 and SKOV-3/DDP cells were treated with cisplatin（DDP） in different concentration for 24h,then the growth inhibiting effects were detected by MTT assay and the inhibitory rate(IC50)was determined by the liner regression of the logarithm of concentration of drugs in order to calculate the resistant index.2 SKOV-3/DDP cells were treated with celecoxib in different concentration for 24h,48h,72h respecitively,then the growth inhibiting effects were detcted by MTT assay and the inhibitory rate（IC10）was determined in order to calculate the non-cytotoxic dose of celecoxib for cells:10μΜ.3 SKOV-3/DDP cells were treated with DDP combinated with celecoxib 10μΜfor 24h,then the growth inhibiting effects were detected by MTT assay and the inhibitory rate(IC50)was determined in order to calculate the reverse fold. 4 To determine the apoptosis effect of SKOV-3/DDP cells after treated with DDP,celecoxib, DDP combinated with celecoxib respecitively for 24h. 5 The intracellular GST-πfluorescence intensity in SKOV-3 and SKOV-3/DDP were measured by flow cytometry（FCM）. 6 To measure the expression of Survivin,GST-πprotein and gene in SKOV-3/DDP cells by flow cytometry（FCM） and RT-PCR respecitively after treated with celecoxib for different time, with DDP, DDP combinated with celecoxib respecitively for 24h.Result:1 MTT assay showed that IC50 of DDP in SKOV-3 and SKOV-3/DDP were 10.119μΜand 27.815μΜrespectively.2 celecoxib could obviously inhibit proliferation of SKOV-3/DDP cells in a dose and time dependent manner.The antiproliferation rates with different concentration celecoxib are 0.564%,4.992%,9.089%,074%,44.104% and 59.323%. The antiproliferation rates treated with 10μΜcelecoxib for 24h,48h,72h, were 9.089%,25.448%,30.635%.there are statistically significant compared with control（P﹤0.01）. The IC10 of celecoxib in SKOV-3/DDP is 8.247μΜ.3 The IC50 of DDP in SKOV-3/DDP cells decrease from 27.815μΜto 15.437μΜ, and the reverse fold is about 1.8 times.4 The apoptotic percentage increased grudually treated with celecoxib,DDP,DDP+celecoxib, and the apoptotic percentage were 7.67%,16.99%,32.75% respecitively, there are statistically significant（P﹤0.01） comparaed with the control group（5.105%）,and there are statistically significant comparaed with DDP group and DDP+celecoxib group（P﹤0.01）.5 there was significantly statistical of the FI-value of GST-πprotein in SKOV-3/DDP and SKOV-3 cells（P﹤0.05）. 6 The FI-value of survivin and GST-πprotein decreaced with celecoxib for 24h,48h,72h.The FI-values of survivin protein were 0.994,0.951,0.945 and The FI-values of GST-πprotein were 0.973,0.941,0.963. there are statistically significant（P﹤0.01） comparaed with the control group.7 The FI-values of survivin protein decreaced with treaed with DDP（0.984）,DDP+celecoxib（0.939）,there are statistically significant（P﹤0.01） comparaed with the control group（0.994） and each other. The FI-values of GST-πprotein with treaed with DDP（1.056）,DDP+celecoxib（0.962）,there are statistically significant（P﹤0.01） comparaed with the control group（0.994） and each other.8 The survivin mRNA decreaced with celecoxib for 24h,48h,72h.The survivin/GAPDH of survivin protein were 0.701,0.518,0.675 , the statistically significant were obviously comparaed with the control group（0.711）.The GST-πmRNA were decreaced with celecoxib for 24h（0.642）,48h（0.463）, but increased for 72h（0.545）,there were statistically significant（P﹤0.05） comparaed with the control group.9 The survivin mRNA decreaced with treaed with DDP（0.496）,DDP+celecoxib（0.34）,there were statistically significant（P﹤0.01） comparaed with the control group and each other. The GST-πmRNA were with treaed with DDP（0.987）,DDP+celecoxib（0.925）,there were statistically significant（P﹤0.01） comparaed with the control group and each other.Conclusion:1 In certain range of concentration,celecoxib could obviously inhibit proliferation of SKOV-3/DDP cells in a dose and time-dependent manner.2 The growth inhibition induced by the combination with celecoxib and DDP present synergistic effects.Celecoxib could enhance the SKOV-3/DDP cells effect of DDP and induce apoptosis in SKOV-3/DDP cells.3 The mechanism of celecoxib reverse SKOV-3/DDP cells’drug resistance may be concerned with down regulation expession of survivin and GST-π.4 This study had showed that celecoxib could be a ideal reversal agent of MDR for going on more studys.

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CLC: > Medicine, health > Oncology > Genitourinary tumors > Female genital tumors > Ovarian tumors
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