目的 探讨活化T细胞核因子c2(NFATc2)对人卵巢癌细胞增殖和凋亡的影响,并研究其可能的机制.方法 选取人上皮性卵巢癌细胞株A2780,分别构建NFATc2过表达(过表达组)和低表达[转染小干扰RNA(siRNA),NFATc2 siRNA组]的A2780细胞株,并设立对照组(不进行任何处理);采用噻唑蓝(MTT)法检测细胞的增殖水平,采用流式细胞仪检测细胞的凋亡水平;采用 Western blot检测凋亡相关蛋白和CXC趋化因子配体8(CXCL8)表达水平.分别用正常的A2780细胞(空白组)、转染空白载体的A2780细胞(对照组)和低表达NFATc2的A2780细胞(NFATc2 siRNA组)建立裸鼠皮下移植瘤的卵巢癌模型,测量肿瘤的体积和质量,用Western blot检测肿瘤组织中CXCL8的表达水平.结果 与对照组比较,NFATc2 siRNA组细胞增殖率降低,细胞凋亡率升高,半胱氨酸蛋白酶3剪切体(cleaved caspase-3)表达水平增加,Bcl-2/Bax比值降低,CXCL8表达水平降低,差异均有统计学意义(P＜0.01);过表达组细胞株则呈相反效应.NFATc2 siRNA组裸鼠肿瘤质量和体积、CXCL8表达水平均明显低于空白组和对照组(P＜0.01).结论 下调NFATc2可通过抑制A2780细胞中CXCL8的表达,减少细胞增殖,增加凋亡相关蛋白表达继而促进卵巢癌细胞凋亡.
Objective To investigate the effect of nuclear factor of activated T cells 2(NFATc2)on the proliferation and apoptosis of human ovarian cancer cells and its possible mechanism.Methods The human epithelial ovarian cancer cell line A2780 was selected,and the A2780 cell lines with over expression(over ex-pression group)and low expression[transfection of small interfering RNA(siRNA),NFATc2 siRNA group]of NFATc2 were constructed,respectively.The control group without any treatment was established.Thiazole blue(MTT)and flow cytometry were used to detect the cell proliferation and apoptosis levels,respectively.The expression levels of apoptosis related proteins and CXC chemokine motif ligand 8(CXCL8)were detected by Western blot.Normal A2780 cells(the blank group),A2780 cells transfected with blank vectors(the con-trol group)and A2780 cells with low expression of NFATc2(the NFATc2 siRNA group)were used to estab-lish ovarian cancer model with subcutaneous xenograft.The tumor volume and mass were measured,and the expression level of CXCL8 in tumor tissue was detected by Western blot.Results Compared with the control group,the cell proliferation rate in the NFATc2 siRNA group was decreased,the cellular apoptosis rate was increased,the cleaved caspase-3 expression level was increased,the Bcl-2/Bax ratio was decreased,the CXCL8 expression was decreased,and the differences were statistically significant(P＜0.01).The cell lines in the o-ver expression group showed the opposite effect.The tumor mass and volume of nude mice,and the expression level of CXCL8 in the NFATc2 siRNA group were lower than those in the blank group and the control group(P＜0.01).Conclusion Down-regulation of NFATc2 could reduce the cell proliferation,increase the expres-sion of apoptosis related proteins by inhibiting CXCL8 expression in A2780 cells,thus promote the apoptosis of ovarian cancer cells.
黄明钜;曾建华
重庆大学附属三峡医院妇科,重庆 404000重庆医科大学附属第二医院妇产科,重庆 400010
临床医学
卵巢肿瘤;活化T细胞核因子;CXC趋化因子配体8;细胞增殖;细胞凋亡
ovarian neoplasms;nuclear factor of activated T cells;CXC chemokine motif ligand 8;pro-liferation;apoptosis
《重庆医学》 2024 (013)
1941-1946,1951 / 7
2022年度重庆市万州区科卫联合医学科研项目(wzstc-kw2022005).
10.3969/j.issn.1671-8348.2024.13.004
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