摘要
目的观察野生型PTEN基因促进乳腺癌多药耐药细胞MCF-7/ADR凋亡的作用并探讨其机制。方法采用脂质体介导法将真核表达载体pEGFP-C1-PTEN及突变载体pEGFP-C1-PTEN-C124S转染人乳腺癌多药耐药MCF-7/ADR细胞。MTT法观察细胞对阿霉素的敏感性和耐药倍数,流式细胞技术检测细胞凋亡率,Western blot检测Bcl-2和Caspase-3蛋白。结果转染组的凋亡率为36.86%,高于对照组(3.75%)和C124S组(4.00%)(P均<0.05)。转染组对阿霉素的耐药倍数显著降低(t=4.77,P<0.05),其半数致死剂量IC50值为对照组的26.1%。转染后细胞内Bcl-2表达降低,且检测到Caspase-3活性裂解片段。结论PTEN基因可能通过下调Bcl-2表达及活化Caspase-3来促进乳腺癌多药耐药MCF-7/ADR细胞凋亡,增加其对阿霉素的药敏性。
Objective To investigate the enhancement effect and mechanism of PTEN on apoptosis induced by adrimytin in breast cancer MCF-7/ADR cell. Methods The eukaryotie expression plasmid pEGFP-CI-PTEN containing whole cDNA of PTEN and its mutation type pEGFP-C1-PTEN-G124S were transfected into MCF-7/ADR cells by Lipofectamine 2000. The apoptosis rate of the cells were observed by flow cytometry. The effect of PTEN on drug chemosensitivity of adrimycin was assessed by MTT tests. The expression of Bel-2 and Caspase-3 were detected by Western blot. Results PTEN overexpression lead to apoptosis of MCF-7/ADR cells （36.86%） detected by flow cytometry, which was much higher than control（3.75% ） and C12S（4.00% ）. The multidrug resistant multiple of the cells with PTEN was also much lower than control（P 〈0.05）. The expression of Bcl-2 transfected with PTEN was significantly decreased and was detected the 17kd active segment of Caspase-3. Conclusion Overexpression of PTEN induces apoptosis through downregulation of Bcl-2 and active caspase3 in human breast cancer MCF-7/ADR ceils.
出处
《山东医药》
CAS
北大核心
2009年第38期19-20,共2页
Shandong Medical Journal
基金
山东省科学技术发展计划项目(2007GG30002024)
