摘要
目的探讨PKM2下调对乳腺癌他莫昔芬(TAM)耐药MCF-7细胞系侵袭、迁移的影响。方法以浓度梯度法诱导获得乳腺癌他莫昔芬耐药细胞系(MCF-7R);细胞增殖与凋亡试剂盒(CCK-8)检测细胞增殖能力;Western blot检测细胞PKM2的表达;将表达针对PKM2的shRNA慢病毒干扰载体感染耐药细胞系,RT-q PCR、Western blot验证干扰效果,分别用Transwell侵袭实验、划痕实验检测细胞侵袭、迁移能力。结果与MCF-7细胞相比,MCF-7R细胞对他莫昔芬的抵抗能力显著增强(P<0.01),PKM2的表达水平明显升高(P<0.01)。稳定干扰PKM2的耐药细胞中PKM2的mRNA和蛋白水平较对照组均明显下降(P<0.01),PKM2干扰后可明显抑制MCF-7R细胞的侵袭、迁移能力(P<0.01)。结论 MCF-7R细胞中PKM2表达明显高于MCF-7细胞,沉默PKM2可以抑制MCF-7R细胞的侵袭和迁移能力。
Objective To investigate the effect of PKM2 down-regulation on invasion and migration in tamoxifenresistant breast cancer cell line MCF-7. Methods The tamoxifen-resistant breast cancer cell line（ MCF-7 R）was established by increasing tamoxifen concentration in a stepwise manner. CCK-8 was used to measure the proliferation of cells. The protein expression of PKM2 was determined by Western blot. shRNA for PKM2 gene lentiviral interference vectors were infected into MCF-7 R cells,the effect of interference was determined by RT-q PCR and Western blot. Cell invasion and migration abilities were measured by Transwell and wound-healing assays.Results The capability of tamoxifen resistance in MCF-7 R cells was dramatically enhanced compared to that in MCF-7 cells（ P〈 0. 01）. Protein level of PKM2 was markedly increased in MCF-7 R cells（ P〈 0. 01）. The mRNA and protein levels of PKM2 declined obviously in PKM2-silented MCF-7R cells（ P〈 0. 01）. Knockdown of PKM2 significantly inhibit the invasion and migration of MCF-7 R cells（ P〈 0. 01）. Conclusions The expressionof PKM2 is apparently higher in MCF-7R than that in MCF-7 cells. Moreover,knockdown of PKM2 can inhibit the migration and invasion of MCF-7R cells.
出处
《基础医学与临床》
CSCD
2016年第12期1641-1645,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(81272544)
