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目的:探讨陵水暗罗提取物暗罗素(ZPT)对耐药卵巢癌细胞株顺铂(DDP)敏感性的影响。方法:MTS法检测暗罗素和顺铂对卵巢癌细胞株A2780和耐药株A2780/DDP的增殖抑制效应。Western Blot法检测A2780和A2780/DDP细胞株p65蛋白表达水平,荧光素酶法检测A2780和A2780/DDP细胞株NF-κB活性。结果:暗罗素可以显著抑制人卵巢癌细胞株A2780和耐药株A2780/DDP的增殖活性[A2780和A2780/DDP的半数抑制浓度(IC50)值分别为1.498μmol/L和1.516μmol/L];联合组(0.5μmol/L暗罗素+5μmol/L顺铂)相对于顺铂单药组,显著下调A2780/DDP的增殖活性(80.80%vs 35.63%,P<0.01)。荧光素酶法检测A2780和A2780/DDP细胞株的NF-κB活性显示,A2780/DDP组NF-κB活性是A2780组的1.92倍(P<0.01);Western Blot法检测p65蛋白表达水平,提示A2780/DDP组p65蛋白表达显著高于A2780组。相对于对照组,0.5μmol/L暗罗素可以显著下调NF-κB活性(100%vs 39.69%,P<0.01)。Western Blot法检查0.5μmol/L暗罗素对A2780/DDP抗凋亡蛋白Bcl-2的影响,发现暗罗素可以显著抑制Bcl-2蛋白表达。结论:陵水暗罗提取物暗罗素可以增加顺铂耐药细胞株A2780/DDP对顺铂的敏感性,暗罗素下调NF-κB-Bcl-2途径活性可能是其逆转顺铂耐药机制。
Objective: To investigate the effect of the crude extract of Lingshui Haliotensin (ZPT) on cisplatin (DDP) sensitivity in drug-resistant ovarian cancer cell lines. Methods: MTS was used to detect the inhibitory effect of dextromethorphan and cisplatin on the proliferation of ovarian cancer cell line A2780 and the resistant strain A2780 / DDP. The protein expression levels of p65 in A2780 and A2780 / DDP cell lines were detected by Western Blot and NF-κB activity in A2780 and A2780 / DDP cell lines by luciferase assay. Results: The dark Russell inhibit the proliferation of human ovarian cancer cell line A2780 and the resistant strain A2780 / DDP [IC50 of A2780 and A2780 / DDP were 1.498μmol / L and 1.516μmol / L, respectively] The combination group (0.5μmol / L dark-Russell + 5μmol / L cisplatin) significantly reduced the proliferation activity of A2780 / DDP (80.80% vs 35.63%, P <0.01) compared with cisplatin alone group. The activity of NF-κB in A2780 and A2780 / DDP cell lines was detected by luciferase assay. The NF-κB activity in A2780 / DDP group was 1.92 times higher than that in A2780 group (P <0.01). The expression of p65 protein was detected by Western Blot, / DDP group p65 protein expression was significantly higher than the A2780 group. Compared with the control group, 0.5μmol / L of dark-Russell significantly reduced NF-κB activity (100% vs 39.69%, P <0.01). The effect of 0.5μmol / L dark-Russell on the anti-apoptotic protein Bcl-2 of A2780 / DDP was detected by Western Blot and found that dark-Russell could significantly inhibit the expression of Bcl-2. It is concluded that the dark-Russell extract of Maitreya can increase the sensitivity of cisplatin-resistant cell line A2780 / DDP to cisplatin. The down-regulation of NF-κB-Bcl-2 pathway by dark Russell may be the mechanism of its reversal of cisplatin resistance.