摘要
[目的]探讨Apogossypolone(ApoG2)对乳腺癌MCF-7细胞株的放射增敏作用及其作用机制。[方法]①MTT法检测ApoG2单药对MCF-7细胞的抑制率,确立细胞半数抑制浓度(IC50)。②根据MTT结果选取低浓度(<IC50)的ApoG2进行克隆形成实验,检测ApoG2对MCF-7细胞的放射增敏作用。③流式细胞仪检测经ApoG2、射线及两者联合作用细胞的凋亡率及自噬荧光强度。④Western Blot检测经ApoG2、射线及两者联合作用细胞的Bcl-2及Be-clin1蛋白表达情况。[结果]ApoG2对乳腺癌细胞株MCF-7细胞的抑制作用随药物浓度的升高,抑制率明显增强,呈一定的剂量依赖性。平板克隆形成实验表明不同浓度ApoG2作用后SF2、Do和Dq均低于照射组。流式细胞技术检测结果显示,与照射组比较,ApoG2与射线联合作用可增加MCF-7细胞凋亡率及自噬荧光强度(P<0.05)。Western Blot结果表明,与照射组比较,ApoG2与射线联合可明显上调Beclin1表达,下调Bcl-2蛋白表达(P<0.05)。[结论]A-poG2对MCF-7乳腺癌细胞株有放射增敏作用,其作用机制不仅与诱导细胞发生凋亡有关,还涉及自噬。自噬可能起到主要抑制作用。
[Purpose ] To investigate ApoG2 enhancing radiosensitivity to MCF-7 breast cancer cell line and its possible mechanism. [ Methods ] ①The inhibition of ApoG2 on MCF-7 cells was exa- mined by MTF assay so as to define IC50.② Low dosage （〈IC50） of ApoG2 based on MTT result was adopted and radiosensitivity of ApoG2 to MCF-7 cell line was determined by clonogenic as- say. ③The apoptosis rate and fluorescence intensity of autophagy in MCF-7 cells treated with ApoG2,irradiation and combination of ApoG2 and irradiation respectively were measured by flow cytometry. ④The expressions of Bcl-2 and Beclinl protein were measured by Western Blot. IRe- sults3 ApoG2 suppressed the proliferation of MCF-7 cells in a dose-time dependent manner. Clonogenic assay showed that in irradiation group, the values of SF2, Do and Dq were higher than those in 2.5μmol/L ApoG2 plus irradiation group and 5μmol/L ApoG2 plus irradiation group. FCM assay indicated that apoptosis rate and fluorescence intensity of autophagy increased in com- bination group compared with those in irradiation group （P〈0.05）. Western Blot showed that the expression of Bcl-2 protein downregulated and the expression of Beclinl upregulated obviously in combination group compared with those in irradiation group（P〈0.05）. [Conclusion] ApoG2 pos- sesses radiosensitizing effect to MCF-7 cells. The mechanism might relate to the induction of cell apoptosis and autophagy, and autophagy plays a major role.
出处
《肿瘤学杂志》
CAS
2012年第8期596-601,共6页
Journal of Chinese Oncology
