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目的探讨4-氨基吡啶(4-AP)对卵巢上皮性癌细胞株SKOV3细胞增殖的抑制作用。方法应用RT-PCR技术分析SKOV3细胞膜上电压门控式钾离子通道(Kv)mRNA的表达;应用膜片钳记录4-AP对SKOV3细胞膜上电压门控式钾离子电流(IK)的抑制作用;应用流式细胞仪检测4-AP对SKOV3细胞增殖周期的影响;采用四甲基偶氮唑蓝(MTT)比色法证实4-AP对SKOV3细胞增殖的抑制作用。以未加4-AP的SKOV3细胞为对照。结果RT-PCR技术检测显示,SKOV3细胞表达KvmRNA;膜片钳记录显示,5mmol/L的4-AP可以降低IK值52·5%;流式细胞仪检测结果显示,应用5mmol/L的4-AP作用72h后,SKOV3细胞周期分布与对照相比,G0/G1期细胞由39·7%增加到62·3%;S期和G2/M期细胞的比例下降,分别由57·3%、3·0%下降到36·2%、1·4%(P<0·05);MTT比色法显示,0·1、1、5、10、15、20mmol/L的4-AP均能抑制SKOV3细胞的增殖,抑制率分别为17·5%、35·0%、54·6%、69·1%、71·2%、72·8%,各浓度分别与对照比较,差异均有统计学意义(P<0·01),且抑制程度与药物浓度呈正相关关系。结论4-AP能够抑制SKOV3细胞的增殖,Kv在卵巢上皮性癌细胞株SKOV3细胞的增殖过程中起着重要作用。
Objective To investigate the inhibitory effect of 4-aminopyridine (4-AP) on the proliferation of ovarian epithelial cancer cell line SKOV3. Methods The expression of voltage-gated potassium channel (Kv) mRNA in SKOV3 cell membrane was analyzed by RT-PCR. The inhibition of voltage-gated potassium current (IK) on SKOV3 cell membrane was recorded by patch clamp. The effect of 4-AP on the proliferation of SKOV3 cells was detected by flow cytometry. The inhibitory effect of 4-AP on the proliferation of SKOV3 cells was confirmed by MTT assay. The SKOV3 cells without 4-AP were used as controls. Results The results of RT-PCR showed that Kv mRNA was expressed in SKOV3 cells. Patch-clamp recording showed that 5 mmol / L 4-AP reduced the IK value by 52.5%. Flow cytometry results showed that 5mmol / L 4- Compared with the control, the cell cycle distribution of SKOV3 increased from 39.7% to 62.3% after 72 h of AP treatment, and decreased from 57.3% in S phase and G2 / M phase, 3 · 0% to 36.2% and 1.4%, respectively (P <0.05). MTT assay showed that the 4-AP of 0 · 1, 1, 5, 10, The inhibition rates of SKOV3 cells were 17.5%, 35.0%, 54.6%, 69.1%, 71.2%, 72.8% respectively. The differences were statistically significant Statistical significance (P <0.01), and the degree of inhibition and drug concentration was positively correlated. Conclusions 4-AP can inhibit the proliferation of SKOV3 cells and Kv plays an important role in the proliferation of SKOV3 cells.