摘要
目的观察细胞分裂周期蛋白42（Cdcd2）在雌激素作用下的变化，探讨细胞内物质运输的改变在雌激素引发的乳腺癌细胞耐药中的意义。方法分别以100ng／ml 17β雌二醇（E2）处理MCF-7细胞，以Cdc,42的小干扰RNA（siRNA）Stealth Select RNAi^TM siRNA转染MCF-7细胞。采用四甲基偶氮唑蓝法检测细胞的药物敏感性，流式细胞术检测细胞内阿霉素（ADM）的蓄积量，实时荧光定量聚合酶链反应检测细胞的Cdc42 mRNA表达，Western blot法检测细胞活化的Cdc42蛋白及总Cdc42蛋白表达量。结果E2处理后，ADM对MCF-7细胞的半数抑制浓度（IC50）由（0．098±0．011）μg/ml增高到（0．134±0．130）μg/ml（P〈0．05），细胞内ADM的相对含量则由7．253±0．310下降为3．233±0．313（P〈0．05），而Cdc42 mRNA、活化Cdc42蛋白及总Cdc42蛋白的表达量均显著增加。（P〈0．05）。Cdc42 siRNA转染后，ADM对MCF-7细胞的IC50下降到（0．057±0．017）μg/ml（P〈0．05），细胞内ADM的相对含量增高为11．217±0．521（P〈0．05），而Cdcd2 mRNA、活化Cdcd2蛋白及总Cdc42蛋白则均有显著下降（P〈0．05）。结论雌激素可以诱导乳腺癌细胞耐药性增强，其机制可能是通过上调Cdc42基因的转录、表达和活化，加速胞内物质运输速度，使得化疗药物无法在细胞内聚集。
Objective To investigate the changes of Cdc42 expression under estrogen stimulation, and to explore the signaling pathway of intracellular material transportation caused by estrogen. Methods MTT was used to test the drug sensitivity of cells. Real-time PCR was used to evaluate the expression of Cclc42 mRNA. The amount of ADM accumulated in MCF-7 cells was detected by flow cytometry. The protein levels of active-Cdc42 and Total-Cdc42 were measured by Western blot. Results ICs0 of ADM in MCF-7 cells was increased from （ 0.098 ± 0.011 ）μg/ml to （ 0. 134 ± 0. 130 ） μg/ml（ P 〈 0.05 ） after estrogen stimulation. The amount of ADM accumulated in MCF-7 cells was reduced from 7. 253 ± 0. 310 to 3. 233 ± 0. 313 （ P 〈 0.05）. All of Cdc42 mRNA, active-Cdc42 protein and total-Cdc42 protein were increased （P 〈 0.05）. After the treatment with siRNA, the ICso of ADM in siRNA group was decreased to （0.057 ±0.017）μg/ml （P 〈 0.05 ） compared with that in the control group. The amount of accumulated ADM was significantly increased in the siRNA group, and all the expression levels of Cdc42 mRNA, active-Cdc42 protein and total- Cdc42 protein were decreased in the siRNA group （P 〈 0.05）. Conclusions Estrogen enhances the drug resistance in breast cancer cells. The mechanism of this effect may be via the enhancing Cdc42 expression and decreasing the accumulation of chemotherapeutic drugs in the cancer cells.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2011年第7期489-493,共5页
Chinese Journal of Oncology
基金
基金项目：国家自然科学基金（30700797）
