丹参酮IIA对子宫内膜癌细胞增殖与凋亡的影响

Effects of tanshinone IIA on the proliferation and apoptosis of endometrial carcinoma cells

目的探讨丹参酮IIA对子宫内膜癌细胞增殖与凋亡的影响。方法将丹参酮IIA作用后的子宫内膜癌细胞设置为对照组、低浓度组(1μg/mL)、中浓度组(2.5μg/mL)及高浓度组(5μg/mL);同时将酪蛋白激酶2(CK2)特异性磷酸化抑制剂(TBB)和丹参酮IIA共同处理后的细胞设置为对照组、TBB组(60μmol)、药物组(5μg/mL)及药物(5μg/mL)+TBB组(60μmol)。结晶紫染色法检测丹参酮IIA对子宫内膜癌细胞增殖的影响;Western blotting检测各组子宫内膜癌细胞中P53、活化胱天蛋白酶-3(Cleaved caspase-3)、B淋巴细胞瘤-2基因(Bcl-2)、CK2α、乳腺癌缺失因子1(DBC1)、沉默信息调节因子1(SIRT1)、磷酸化乳腺癌缺失因子1(p-DBC1)的表达;同时加入TBB,检测子宫内膜癌细胞中CK2α、DBC1、p-DBC1的表达。结果结晶紫染色结果显示,对照组和高浓度组各时间点(24、48、72 h)细胞增殖率分别为(0.87±0.05)%、(0.61±0.04)%、(0.30±0.04)%、(0.09±0.04)%,差异有统计学意义(F=217.976,P<0.001)。Western blotting结果显示,在丹参酮IIA作用下,P53(F=244.261)及Cleaved caspase-3(F=290.842)的表达水平呈浓度依赖性升高,Bcl-2(F=198.170)的表达水平呈浓度依赖性降低,差异均有统计学意义(P均<0.001);p-DBC1/DBC1的表达水平呈浓度依赖性降低,差异有统计学意义(F=187.092,P<0.001),而CK2α、SIRT1的表达差异均无统计学意义(P均>0.05);单独应用TBB(F=7.847)、丹参酮IIA(F=827.715)及联合应用(F=22.174),p-DBC1/DBC1表达均受到抑制,与对照组比较,差异均有统计学意义(P=0.023,P<0.001,P=0.002);CK2α表达差异无统计学意义(P>0.05)。结论丹参酮IIA可以明显抑制子宫内膜癌细胞增殖并诱导凋亡,其机制可能与CK2/DBC1/SIRT1/P53信号通路有关。

Objective To investigate the effects of tanshinone IIA on the proliferation and apoptosis of endometrial carcinoma cells.Methods According to the experimental requirements,endometrial carcinoma cells treated with tanshinone IIA were divided into the control group,low concentration group(1μg/mL),medium concentration group(2.5μg/mL)and high concentration group(5μg/mL).Meanwhile,cells co-treated with CK2 specific phosphorylation inhibitor(TBB)and tanshinone IIA were divided into the control group,TBB group(60μmol),drug group(5μg/mL)and drug(5μg/mL)+TBB group(60μmol).The effects of tanshinone IIA on the proliferation of cells were detected with crystal violet staining;expressions of P53,Cleaved caspase-3,B-cell lymphoma-2(Bcl-2),casein kinase 2α(CK2α),deletion in breast cancer 1(DBC1),silent information regulator 1(SIRT1)and p-deletion in breast cancer 1(p-DBC1)were detected with Western blotting.After TBB was added,expressions of CK2α,DBC1 and p-DBC1 were also detected.Results The results of crystal violet staining showed that the proliferation rates of the control group and high concentration group at each time point(24 h,48 h,72 h)were(0.87±0.05)%,(0.61±0.04)%,(0.30±0.04)%and(0.09±0.04)%,respectively(F=217.976,P<0.001).The results of Western blotting showed that under the action of tanshinone IIA,the expressions of P53(F=244.261)and Cleaved caspase-3(F=290.842)increased in a concentration-dependent manner,while the expression of Bcl-2(F=198.170)decreased in a concentration-dependent manner(P<0.001),and the expression of p-DBC1/DBC1 decreased in a concentration-dependent manner(F=187.092,P<0.001),but there was no significant difference in CK2αand SIRT1 expressions(P>0.05).The expression of p-DBC1/DBC1 was inhibited by TBB(F=7.847),tanshinone IIA(F=827.715)and their combination(F=22.174),and the difference was statistically significant compared with the control group(P=0.023,P<0.001,P=0.002),but there was no significant difference in CK2αexpression(P>0.05).Conclusion Tanshinone IIA can significantly inhibit the proliferation and induce apoptosis of endometrial carcinoma cells,and the mechanism may be related to the CK2/DBC1/SIRT1/P53 signaling pathway.