摘要
目的探讨miR-1285通过转录共激活因子Yes相关蛋白1(YAP1)对慢性粒细胞白血病K562细胞增殖、凋亡的影响及其可能的作用机制。方法构建miR-1285的质粒,使K562细胞过表达miR-1285及敲低miR-1285,分为miR-1285 mimics、mimics control、miR-1285 inhibitor、inhibitor control四组;采用qRT-PCR法定量分析四组中miR-1285的表达量,验证转染效率,利用CCK8、AnnexinⅤ-FITC方法检测四组K562细胞的凋亡、增殖情况;使用Western blot检测YAP及其下游分子表皮生长因子受体(EGFR)等变化,检测凋亡相关分子BAX、Bcl-2蛋白表达变化。结果敲低miR-1285后,可促进K562细胞的增殖,抑制凋亡,YAP的表达升高,下游分子EGFR表达相应升高,凋亡相关分子BAX降低,而Bcl-2表达升高;而过表达miR-1285后,K562细胞增殖减弱,凋亡增加,YAP表达降低,下游分子P-EGFR表达降低,凋亡分子BAX表达升高,而Bcl-2表达降低。结论miR-1285可通过抑制YAP的表达从而抑制慢性粒细胞白血病K562细胞的增殖、促进凋亡,有望成为治疗慢性粒细胞白血病的靶向分子。
Objective To study the miR-1285 through Yes-associated protein 1(YAP1)on the proliferation,apoptosis and mechanism of chronic myelogenous leukemia K562 cells.Methods The plasmid of miR-1285 was constructed to make K562 cells overexpress miR-1285 and knock down miR-1285.They were divided into four groups:miR-1285 mimics,mimics control,miR-1285 inhibitor,and inhibitor control;qRT-PCR was used to quantitatively analyze the expression of miR-1285 of these four groups.The expression level of miR-1285 in the group was used to verify the transfection efficiency.The apoptosis and proliferation of the four groups were detected by CCK-8 and Annexin V-FITC;the changes of YAP and its downstream molecule epidermal growth factor receptor(EGFR)were detected by Western blot,and apoptosis related molecules BAX,Bcl-2 protein expression was detected.Results After knock down miR-1285,it can promote the proliferation of K562 cells and inhibit apoptosis.The expression of YAP increased,the expression of downstream molecule EGFR increased correspondingly,the expression of apoptosis-related molecule BAX decreased,and the expression of Bcl-2 increased;After expressing miR-1285,K562 cell proliferation decreased,apoptosis increased,YAP expression decreased,downstream molecule P-EGFR expression decreased,apoptosis molecule BAX expression increased,and Bcl-2 expression decreased.Conclusion miR-1285 can inhibit the proliferation and promote apoptosis of K562 cells of chronic myelogenous leukemia by inhibiting the expression of YAP.It is expected to become a targeted molecule for the treatment of chronic myelogenous leukemia.
作者
张红霞
吴广胜
Zhang Hongxia;Wu Guangsheng(Dept of Hematology,The First Affiliated Hospital of Shihezi University Medical College,Shihezi 832000)
出处
《安徽医科大学学报》
CAS
北大核心
2022年第4期569-573,共5页
Acta Universitatis Medicinalis Anhui
基金
石河子大学2021年兵团财政科技计划项目(编号:2021CA002)
石河子大学2021年度自主资助支持校级科研项目(编号:ZZZC202186)。
