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下调钙网蛋白表达对乳腺癌干细胞增殖、侵袭和凋亡的影响
Effects of downregulated calreticulin on the proliferation,invasion and apoptosis of breast cancer stem cells
【摘要】 目的探讨钙网蛋白(calreticulin,CRT)对乳腺癌干细胞增殖、侵袭和凋亡能力的影响。方法采用流式细胞分选技术从人乳腺癌细胞系MDA-MB-231中分离出乳腺癌干细胞。稳定转染重组质粒pGPU6/CRT(shRNA-CRT),Western blotting法检测乳腺癌干细胞CRT蛋白的表达,MTT法和transwell法分别检测pGPU6/CRT对乳腺癌干细胞增殖和侵袭能力的影响,AnnexinⅤ-FITC/PI双染法检测pGPU6/CRT对乳腺癌干细胞凋亡的影响。结果从MDA-MB-231细胞系中成功分选出乳腺癌干细胞;pGPU6/CRT转染乳腺癌干细胞后,CRT的表达水平显著降低;下调的CRT表达能够显著抑制乳腺癌干细胞增殖和侵袭能力,同时诱导乳腺癌干细胞凋亡。结论下调CRT能够抑制乳腺癌干细胞的增殖和侵袭能力,并促进干细胞凋亡。
【Abstract】 Objective To investigate the effects of calreticulin (CRT) on the proliferation,invasion and apoptosis of breast cancer stem cells.Methods Breast cancer stem cells were separated from human breast cancer cell line MDA-MB-231 by flow cytometry.The breast cancer stem cells were stably transfected into the recombinant plasmid pGPU6/CRT (shRNA-CRT).Then Western blotting assay was adopted to identify the expression of CRT.The effects of pGPU6/CRT on the proliferation and invasion of breast cancer stem cells were detected by MTT and transwell assay respectively.The effect of pGPU6/CRT on the apoptosis of breast cancer stem cells was detected with Annexin Ⅴ-FITC/PI double staining flow cytometry.Results Breast cancer stem cells were successfully separated from human breast cancer cell line MDA-MB-231.The significant downregulation of CRT expression caused by transfection of pGPU6/CRT into breast cancer stem cells was detected by Western blotting.The ability of proliferation and invasion was inhibited successfully by the downregulated expression of CRT,and apoptosis was induced in breast cancer stem cells as well.Conclusion The downregulated expression of CRT could inhibit the proliferation and invasion of breast cancer stem cells and promote apoptosis.
【Key words】 human breast cancer stem cell; calreticulin; proliferation; invasion; apoptosis;
- 【文献出处】 解剖科学进展 ,Progress of Anatomical Sciences , 编辑部邮箱 ,2013年04期
- 【分类号】R737.9
- 【被引频次】8
- 【下载频次】155