槲皮素对人卵巢癌细胞-3增殖和凋亡的影响

Effect of Quercetin on the Apoptosis and Proliferation of Human Ovarian Cancer Cell Strainon OVCAR-3 Cells

目的:探讨槲皮素对人卵巢癌细胞-3(ovarian carcinoma cell,OVCAR-3)增殖和凋亡的影响及其作用机制。方法:将对数生长期的OVCAR-3细胞随机分为对照组和槲皮素组。对照组细胞给予培养基常规培养,而槲皮素组细胞加入槲皮素(160μmol·L^(-1))。各组细胞培养48 h后,利用噻唑蓝检测人卵巢癌细胞株OVCAR-3细胞增殖;流式细胞仪检测人卵巢癌细胞株OVCAR-3细胞凋亡,RT-PCR检测细胞受体型酪氨酸激酶2(JAK2),信号传导与转录激活因3(STAT3),B细胞淋巴因子2(Bcl-2),人Bcl-2相关X蛋白(Bax)信使mRNA表达水平。免疫蛋白印记法检测各组OVCAR-3细胞中STAT3,磷酸化受体型酪氨酸激酶2(p-JAK2),磷酸化信号传导与转录激活因3(p-STAT3),Bax,Bcl-2蛋白的变化,Caspase3活性试剂盒检测Caspase3活性。结果:MTT检测结果显示槲皮素组与对照组相比,人卵巢癌细胞株OVCAR-3细胞增殖率明显下降(P<0.05)。流式细胞仪检测显示槲皮素组OVCAR-3细胞凋亡率较对照组明显升高(P<0.05)。RT-PCR检测显示槲皮素组与对照组JAK2和STAT3信使mRNA表达水平无明显差异,而槲皮素组Bax信使mRNA水平比对照组明显增高,槲皮素组Bcl-2信使mRNA水平比显著低于对照组。Western blotting检测显示,槲皮素组的JAK2和STAT3蛋白表达与对照组比较,无明显差异(P>0.05),而槲皮素组的p-STAT3,p-JAK2和Bcl-2蛋白表达水平显著低于对照组(P<0.05),槲皮素组的Bax表达显著高于对照组(P<0.05)。Caspase3活性试剂盒检测结果显示,槲皮素组Caspase3活性表达水平显著高于对照组。结论:槲皮素能明显抑制人卵巢癌细胞株OVCAR-3细胞增殖,促进人卵巢癌细胞株OVCAR-3细胞凋亡,其作用机制可能是通过激活JAK2/STAT3信号传导通路。

Objective： To investigate the effect and potential mechanisms of the Quercetin on the human ovarian cancer cell strainon OVCAR-3 cells. Methods： The OVCAR-3 cells at logarithmic growth phase were randomly divided into control and quercetin groups. The cells in the control group were normally treated with culture medium and the cells in the quercetin group were incubated with quercetin at the dose of 160 μmol·L^（-1）. After the cell is two groups were treated with 48 h,the cell was harvest to analysis. The proliferation rate of human ovarian cancer cell strainon OVCAR-3 cells was examined by 3-（ 4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（ MTT）. The apoptosis rats of OVCAR-3 cells in two groups was measured by Flow cytometry. The mRNA level of Janus activating Kinase 2（ JAK2）,signal transducer and activator of transcription3（ STAT3）,Bcl-2 associated X protein（ Bax） and B-cell lymphoma-2（ Bcl-2） were detected by RT-PCR. The protein expression level of Janus activating Kinase 2（ JAK2）,phosphorylation of Janus activating kinase 2（ p-JAK2）,signal transducer and activator of transcription 3（ STAT3）,phosphorylation of signal transducer and activator of transcription3（ p-STAT3）,Bcl-2 associated X protein（ Bax） and B-cell lymphoma-2（ Bcl-2） were measured by Western blotting. The activity of Caspase3 was measured by apoptotic protease3（ caspase3） colorimetric assay kit. Results： Compared with the control group,the proliferation rate of OVCAR-3 cells in quercetin group was significantly decreased P 〈0. 05）,but the apoptotic rate of cells in quercetin group was significantly increased（ P 〈0. 05）. After the treatment with quercetin,the mRNA expression level of Bax in the quercetin group was higher than control group（ P 〈0. 05）. Furthermore,the mRNA expression level of BCL-2 in the quercetin group was lower than control group（ P 〈0. 05）. However,there was no difference on the mRNA level of JAK2 and STAT3 between two groups（ P 〈0. 05）. The western blotting results showed that the protein expression level of p-JAK2,p-STATA3 and Bax in the quercetin group were significantly higher than the control group（ P 〈0. 05）. Furthermore,the protein expression level of BCL-2 in the quercetin group was lower than control group（ P 〈0. 05）. However,there was no significant difference on the protein expression level of total JAK2 and STAT3 protein between two groups（ P 〈0. 05）. The apoptotic protease 3（ caspase3） colorimetric assay kit showed that compared with the control group,the activity of Caspase3 in quercetin group was significantly higher（ P 〈0. 05）. Conclusion： Quercetin could significantly inhibited the proliferation of human ovarian cancer cell strainon OVCAR-3 cells and induced the apoptosis of OVCAR-3 cell. The effect of quercetin on the OVCAR-3 cells may be achieved through promoting JAK2 / STAT3 signal transduction pathway activation.