摘要
目的探讨木犀草素联合下调miR-425-5p对宫颈癌HeLa细胞生物学特性的影响及可能机制。方法将宫颈癌HeLa细胞(购自中国医学科学院肿瘤细胞库)分成Control组(不做任何处理)、Anti-NC组(在HeLa细胞中用Lipofectamine 2000转染inhibitor control)、Anti-miR-425-5p组(在HeLa细胞中用Lipofectamine 2000转染miR-425-5p inhibitor)、木犀草素+Anti-NC组(在HeLa细胞中用Lipofectamine 2000转染inhibitor control,经40μmol/L木犀草素处理)、木犀草素+Anti-miR-425-5p组(在HeLa细胞中用Lipofectamine 2000转染miR-425-5p inhibitor,经40μmol/L木犀草素处理)共5组,并对上述5组采用MTT测定细胞培养24 h后A值(以A值代表细胞增殖活性),平板克隆实验测定细胞培养12 d后克隆形成率,流式细胞术检测细胞培养24 h后凋亡率,Transwell小室测定细胞培养24 h后侵袭和迁移数目,Western blot法检测细胞培养24 h后p-Akt、c-caspase-3、c-caspase-9、基质金属蛋白酶-9(MMP-9)、MMP-2、Bax、Bcl-2蛋白表达水平,比较5组上述指标差异。结果与Anti-NC组比较,Anti-miR-425-5p组、木犀草素+Anti-NC组、木犀草素+Anti-miR-425-5p组细胞增殖活性、侵袭与迁移数目和MMP-9、MMP-2、p-Akt蛋白表达水平均明显降低,细胞凋亡率和c-caspase-3、c-caspase-9、Bax蛋白表达水平升高,Bcl-2蛋白表达减少,差异均有统计学意义(P<0.05)。与Anti-miR-425-5p组、木犀草素+Anti-NC组比较,木犀草素+Anti-miR-425-5p组细胞增殖活性、侵袭与迁移数目和MMP-9、MMP-2、p-Akt蛋白表达水平均明显降低,细胞凋亡率和c-caspase-3、c-caspase-9、Bax蛋白表达水平升高,Bcl-2蛋白表达减少,差异均有统计学意义(P<0.05)。结论木犀草素联合下调miR-425-5p可抑制宫颈癌HeLa细胞增殖、侵袭和迁移,诱导细胞凋亡,作用机制可能与抑制Akt信号通路的激活有关。
Objective To investigate the effect of luteolin combined with down-regulation of miR-425-5p on the biological characteristics of cervical cancer cellsand the underlying mechanism.Methods Cervical cancer HeLa cells(purchased from the Tumor Cell Bank of the Chinese Academy of Medical Sciences)were divided into control group(no treatment),anti-NC group(inhibitor control transfected with Lipofectamine 2000 in HeLa cells),anti-miR-425-5p group(miR-425-5p inhibitor transfected with Lipofectamine 2000 in HeLa cells),Luteolin+anti-NC group(inhibitor controltransfected with Lipofectamine 2000 in HeLa cells,treated with 40μmol/L luteolin),Luteolin+anti-miR-425-5p group(miR-425-5p inhibitor transfected with Lipofectamine 2000 in HeLa cells,treated with 40μmol/L luteolin),a total of five groups,and the A value of cells after 24 hours of culture(the A value represents cell proliferation activity)inthe abovefive groups were determined by MTT.The plate cloning experiment was used to measure the rate of clone formation after 12 days of culture,the flow cytometry was employed to detect the rate of cell apoptosis after 24 hours of culture,the Transwell chamber was appliedto measure cell invasion and migration after 24 hours of culture,and Western blot was used to detect p-Akt,c-Caspase-3,c-Caspase-9,MMP-9,MMP-2,Bax,Bcl-2 protein expression levels in cells after 24 hours of culture.Results Compared with anti-NC group,the cell proliferation activity,invasion number,migration number and MMP-9,MMP-2,p-Akt protein in anti-miR-425-5p,luteolin+anti-NC,luteolin+anti-miR-425-5p group were significantly reduced,the rate of apoptosis and the levels of c-Caspase-3,c-Caspase-9,and Bax protein increased,and the expression of Bcl-2 protein decreased,the difference being statistically significant(P<0.05).Compared with anti-miR-425-5p,luteolin+anti-NC group,the cell proliferation activity,invasion number,migration number and MMP-9,MMP-2,p-Akt in luteolin+Anti-miR-425-5p group were significantly reduced,the apoptotic rate and the levels of c-Caspase-3,c-Caspase-9,and Bax protein increased,and the expression of Bcl-2 protein decreased,the difference being statistically significant(P<0.05).Conclusions Luteolin combined with down-regulation of miR-425-5p could inhibit the growth,invasion and migration of cervical cancer cells,and induce cell apoptosis.The mechanism of action may be related to the inhibition of the activation of Akt signaling pathway.
作者
王学博
符攀峰
任红娟
孙丽
白峰岩
WANG Xuebo;FU Panfeng;REN Hongjuan(Department of Gynecology,Nanyang Central Hospital,Nanyang 473000,China)
出处
《安徽医学》
2021年第2期119-123,共5页
Anhui Medical Journal
