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目的:观察醋酸甲羟孕酮(MPA)对人卵巢癌CoC1/cDDP细胞移植瘤的生长抑制作用及对DDP的耐药逆转作用,并分析其作用机制。方法:建立人卵巢癌裸鼠皮下移植瘤模型,随机分为4组,每组5只。(1)对照组:腹腔注射等体积生理盐水;(2)DDP治疗组:每只每次腹腔注射DDP 3mg/kg。(3)MPA治疗组:每只每次灌胃30mg/kg;(4)联合治疗组:每只每次灌胃MPA 30mg/kg,1h后每只每次腹腔注射DDP 3mg/kg,每隔2天给药1次,共4次,于治疗第1、5、10、15、20天分别测瘤体积,20天后处死裸鼠,完整剥出瘤组织,称瘤重,计算抑瘤率;通过流式细胞仪检测亚G1期细胞及AnnexinV+/PI-细胞鉴定细胞凋亡,分析移植瘤细胞周期;用半定量RT-PCR法检测移植细胞组织Survivin-ΔEx3、caspase-3、P21WAF1/CIP1及GST-π4基因mRNA表达。结果:(1)MPA组、MPA+DDP组治疗第10天起皮下移植瘤体积明显小于DDP组和对照组,且进行性缩小(P<0.01);抑瘤率分别为51.63%、62.21%,均明显大于DDP组的6.84%(P<0.01),并且两组间有显著差异(P<0.01);(2)流式细胞仪分析显示,移植瘤出现亚G1期峰及AnnexinV+/PI-细胞均证实MPA能诱导CoC1/cDDP细胞凋亡,并显著高于对照组及DDP组,并出现G1期阻滞;与DDP合用除出现G1期阻滞外又出现G2/M期阻滞,S期明显减少,亚G1期细胞及AnnexinV+/PI-细胞进一步上升;(3)半定量RT-PCR检测显示,MPA组Survivin-ΔEx3、GST-πmRNA下调,而P21WAF1/CIP1、caspase-3 mRNA上调,与DDP合用后,对Survivin-ΔEx3、P21WAF1/CIP1及GST-πmRNA的表达无协调作用,而对caspase-3 mRNA有协同上调作用。结论:MPA通过阻滞G0/G1期明显抑制了CoC1/cDDP移植瘤生长,并有很强的致凋亡作用,同时下调GST-πmRNA,从而逆转对顺铂耐药。
Objective: To observe the inhibitory effect of medroxyprogesterone acetate (MPA) on the growth of human ovarian cancer CoC1 / cDDP cell xenografts and the drug resistance reversal effect on DDP, and to analyze its mechanism of action. Methods: Human ovarian cancer xenograft model of subcutaneous xenografts was established and randomly divided into 4 groups with 5 in each group. (1) control group: intraperitoneal injection of equal volume of saline; (2) DDP treatment group: each intraperitoneal injection of DDP 3mg / kg. (3) MPA treatment group: each time each gavage 30mg / kg; (4) combined treatment group: Each time each gavage MPA 30mg / kg, 1h after each intraperitoneal injection of DDP 3mg / kg, every 2 days for 1 time, a total of 4 times on the 1st, 5th, 10th, 15th and 20th days respectively. The nude mice were sacrificed after 20 days, and the tumor tissues were completely stripped and weighed to calculate the tumor inhibition rate. Cell apoptosis was detected by flow cytometry in sub-G1 phase and Annexin V + / PI-cells, and the cell cycle was analyzed. Survivin-ΔEx3, caspase-3, P21WAF1 / CIP1 GST-π4 mRNA expression. Results: (1) The volume of subcutaneously transplanted tumor in MPA group and MPA + DDP group was significantly smaller than that in DDP group and control group on the 10th day after treatment (P <0.01), and the inhibition rates were 51.63%, 62.21% (P <0.01). (2) Flow cytometry analysis showed that sub-G1 peak and Annexin V + / PI-cells MPA could induce the apoptosis of CoC1 / cDDP cells and was significantly higher than the control group and the DDP group, and G1 arrest; combined with DDP in addition to G1 arrest appears again G2 / M phase arrest, S phase (3) Semi-quantitative RT-PCR showed that downregulation of Survivin-ΔEx3 and GST-πmRNA, and up-regulation of P21WAF1 / CIP1 and caspase-3 mRNA in MPA group were associated with DDP combined with no effect on the expression of Survivin-ΔEx3, P21WAF1 / CIP1 and GST-πmRNA, but synergistic up-regulation of caspase-3 mRNA. CONCLUSION: MPA can significantly inhibit the growth of CoC1 / cDDP xenografts by arresting the G0 / G1 phase and have a strong apoptosis-inducing effect. At the same time, MPA down-regulates GST-πmRNA and reverse the resistance to cisplatin.