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目的 观察特异性热休克蛋白70(heatshockprotein,HSP)反义寡核苷酸阻断卵巢癌细胞的HSP70表达,及其对卵巢癌细胞生长和增殖的影响。方法 用胎盘蓝拒染法计算卵巢癌细胞的生长抑制率,Giemsa染色法从形态上了解凋亡的发生,用琼脂糖凝胶电泳进一步检测发生凋亡的特征性DNA降解,流式细胞仪定量分析凋亡发生率及周期特异性。结果 用HSP70反义寡核苷酸处理的卵巢癌细胞表现出明显的生长抑制,在形态学上显示出细胞凋亡的特征,琼脂糖凝胶电泳显示凋亡细胞DNA降解的特征性DNA阶梯图。流式细胞仪检测表明1～20μmolL的反义寡核苷酸能诱导83%～41%的卵巢癌细胞发生凋亡,主要是G1期和S期细胞,凋亡发生随药物剂量及药物作用时间增加而增多。结论 HSP70反义寡核苷酸能抑制卵巢癌细胞的生长并诱导其发生凋亡。
Objective To observe the effect of heatshock protein (HSP) antisense oligonucleotides on the expression of HSP70 in ovarian cancer cells and its effect on the growth and proliferation of ovarian cancer cells. Methods The growth inhibition rate of ovarian cancer cells was calculated using placenta blue exclusion method. The appearance of apoptosis was morphologically understood by Giemsa staining. The characteristic DNA degradation with apoptosis was further detected by agarose gel electrophoresis, and the flow cytometry was used for quantitative analysis. Analysis of the incidence of apoptosis and cycle specificity. Results The ovarian cancer cells treated with HSP70 antisense oligonucleotide exhibited obvious growth inhibition, showed morphological characteristics of apoptosis, and agarose gel electrophoresis showed a characteristic DNA ladder diagram of DNA degradation of apoptotic cells. . Flow cytometry showed that 1～20μmol L antisense oligodeoxynucleotide could induce apoptosis of 83%～41% of ovarian cancer cells, mainly G1 and S phase cells, apoptosis occurred with drug dosage and drug action time. Increase and increase. Conclusion HSP70 antisense oligonucleotide can inhibit the growth of ovarian cancer cells and induce apoptosis.