腺苷酸活化蛋白激酶介导多柔比星诱导的抗人类乳腺癌MCF-7细胞增殖作用

Activation of AMP-activated protein kinase is involved in doxorubicin-induced anti-breast cancer cell proliferation

目的研究多柔比星对乳腺癌MCF．7细胞增殖的影响，并探讨腺苷酸活化蛋白激酶（AMPK）在其中的作用。方法分别用AMPK激活剂AICAR、siRNA敲低AMPK表达（AMPKsiRNA）、AMPK抑制剂复合物C（AMPKi）联合多柔比星（DOX）对MCF-7细胞进行处理，在不同时间通过Westernblot方法检测AMPK、乙酰辅酶A羧化酶（ACC）、p38的活化，MTF检测细胞存活率间接反应细胞增殖。结果DOX可诱导MCF-7细胞AMPK的活化，AICAR单独或联合DOX可诱导AMPK的激活及增加MCF-7细胞增殖抑制率，AICAR＋DOX组与DOX组细胞存活率分别为（17．74-1．6）％和（71．4±1．8）％（P〈0．001）；AMPKi或AMPKsiRNA联合DOX后，P—AMPK及P—ACC表达明显下降，p38活化水平不受影响，MCF-7细胞增殖抑制率下降，AMPKi＋DOX组与DOX组细胞存活率分别为（72．7±1．8）％和（96-3±1．7）％（P〈0．001），AMPKsiRNA＋DOX组与错义siRNA＋DOX组细胞存活率分别为（76．9±2．2）％和（95．9±1．8）％（P〈0．001）。结论AMPK介导DOX诱导的抗乳腺癌细胞增殖。

Objective To investigate the mechanism of anti-breast cancer cell proliferation induced by doxorubicin （DOX）. Methods AMP-Activated Protein Kinase （AMPK） activator AICAR, AMPK siRNA, AMPK inhibitor compound C （AMPKi） and doxorubiein treated MCF-7 cells at different time points; AMPK, acetyl CoA carboxylase （ACC）, p38 activation were detected by Western blot. MTY was used as cell viability assay. Results Doxorubicin-induced activation of AMPK, AMPK agonist （AICAR）or in combination with doxorubicin activated AMPK and increased MCF-7 cell proliferation rate [the difference of cell viability between group AICAR±DOX（17.7±1.6 ） % and group DOX（71.4±1.8 ） % was significant（P〈0.001）]. After AMPKi or AMPK siRNA and doxorubicin combined administration, P-AMPK and P-ACC expression was significantly decreased, the level of p38 was not affected, and MCF-7 cell proliferation inhibition rate decreased [the cell viability of group AMPKi±DOX（72.7±1.8 ） % vs group DOX（96.3±1.7 ） %,P〈0.001;group AMPK siRNA ±DOX（ 76.9±2.2 ） % vs group scramble siRNA±DOX（95.9±1.8） %,P〈0.001]. Conclusion AMPK is involved in doxorubicin-indueed anti-breast cancer cell proliferation.