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目的:探讨丙氨瑞林对卵巢癌细胞株CoC1/cDDP增殖及凋亡的影响及其作用机制。方法:将丙氨瑞林与CoC1/cDDP一起培养,采用四甲基偶氮唑蓝(MTT)比色法检测细胞增殖抑制率,流式细胞仪进行细胞周期时相及凋亡分析,瑞氏-姬姆萨染色后观察凋亡细胞形态,放射免疫法测定培养液中CA125的变化,半定量RT-PCR法检测作用前后CoC1/cDDP细胞Survivin-ΔEx3及Caspase-3 mRNA的表达。结果:(1)随着丙氨瑞林浓度的升高,作用时间的延长,细胞生长抑制率逐渐上升(P<0.01)。(2)丙氨瑞林作用后的CoC1/cDDP细胞,G0/G1期细胞比例增加,S期及G2/M期细胞比例减少,细胞增殖指数(PI)降低(P<0.01);培养液中CA125的水平下降,与PI正相关(r=0.893,P<0.05);并且这些变化与丙氨瑞林的浓度相关。(3)作用后亚G1期及Annexin V+/PI-细胞的百分率较对照组明显升高(P<0.01),并出现明显的凋亡细胞形态的改变;Survivin-ΔEx3 mRNA表达与对照组比较无明显变化(P>0.05),而Caspase-3 mRNA的表达上调,并随着丙氨瑞林浓度的升高而表达增加(P<0.01),且与亚G1期及Annexin V+/PI-细胞的比例呈正相关(r分别为0.994及0.978,P<0.01)。结论:丙氨瑞林能直接抑制卵巢癌细胞株CoC1/cDDP细胞的增殖,并诱导其凋亡,作用机制与丙氨瑞林上调Caspase-3 mRNA的表达有关。
Objective: To investigate the effect and mechanism of ararelin on the proliferation and apoptosis of ovarian cancer cell line CoC1 / cDDP. METHODS: Ammunorelin was co-cultured with CoC1 / cDDP. MTT assay was used to detect the cell proliferation inhibition rate, cell cycle phase and apoptosis analysis by flow cytometry. The morphology of apoptotic cells was observed after Giemsa staining. The expression of CA125 in culture medium was detected by radioimmunoassay. The expression of Survivin-ΔEx3 and Caspase-3 mRNA in CoC1 / cDDP cells were detected by semi-quantitative RT-PCR. Results: (1) As the concentration of ararelin increased, the prolongation of action time, the cell growth inhibition rate increased gradually (P <0.01). (2) The percentage of cells in G0 / G1 phase and the proportion of cells in S phase and G2 / M phase were decreased and the cell proliferation index (PI) was decreased (P <0.01) after treated with ararelin. CoC1 / The level of CA125 decreased and was positively correlated with PI (r = 0.893, P <0.05); and these changes correlated with the concentration of ararelin. (3) After treatment, the percentage of sub-G1 phase and Annexin V + / PI-cells was significantly higher than that of the control group (P <0.01), and the morphological changes of apoptotic cells were obvious. Survivin-ΔEx3 mRNA expression was lower than that of the control group (P> 0.05), while the expression of Caspase-3 mRNA was up-regulated and increased with the increase of the concentration of the arareverin (P <0.01). Compared with the sub-G1 and Annexin V + / PI- The proportions were positively correlated (r = 0.994 and 0.978, respectively, P <0.01). CONCLUSIONS: Alarelin can directly inhibit the proliferation and induce the apoptosis of ovarian cancer cell line CoC1 / cDDP. The mechanism of action is related to the up-regulation of Caspase-3 mRNA expression by ararelin.