沉默PKM2对乳腺癌他莫昔芬耐药MCF-7细胞系侵袭及迁移的影响

基础医学与临床 ›› 2016, Vol. 36 ›› Issue (12): 1641-1645.

沉默PKM2对乳腺癌他莫昔芬耐药MCF-7细胞系侵袭及迁移的影响

季飞虎,王念,钟昌莉,蒋玉林,刘一锋,张志乾,靳倩妮,陈婷梅

重庆医科大学

收稿日期:2016-05-19 修回日期:2016-09-25 出版日期:2016-12-05 发布日期:2016-11-29
通讯作者: 陈婷梅 E-mail:chentingmei@sohu.com
基金资助:
国家自然科学基金

Influence of PKM2 silencing on invasion and migration in tamoxifen-resistant breast cancer cell line MCF-7

Received:2016-05-19 Revised:2016-09-25 Online:2016-12-05 Published:2016-11-29
Supported by:
Natural Science Foundation of China

摘要/Abstract

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-qPCR、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。

关键词: 乳腺肿瘤, 他莫昔芬, M2-型丙酮酸激酶, 侵袭, 迁移

Abstract: Objective To investigate the effect of PKM2 down-regulation on invasion and migration in tamoxifen-resistant breast cancer MCF-7 cell line. Methods The tamoxifen-resistant breast cancer cell line (MCF-7R) 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-7R cells, the effect of interference was determined by RT-qPCR and Western blot. Cell invasion and migration abilities were measured by transwell and wound-healing assays. Results The capability of tamoxifen resistance in MCF-7R cells was dramatically enhanced compared to that in MCF-7 cells (P<0.01). Protein level of PKM2 was markedly increased in MCF-7R cells (P<0.01). The mRNA and protein levels of PKM2 declined obviously in PKM2-silented MCF-7R cells (P<0.01). We also demonstrated that knockdown of PKM2 significantly inhibit the invasion and migration of MCF-7R cells (P<0.01). Conclusions The expression of 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.

Key words: Breast cancer, Tamoxifen, pyruvate kinase M2, invasion, migration

季飞虎王念钟昌莉蒋玉林刘一锋张志乾靳倩妮陈婷梅. 沉默PKM2对乳腺癌他莫昔芬耐药MCF-7细胞系侵袭及迁移的影响[J]. 基础医学与临床, 2016, 36(12): 1641-1645.

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