目的研究S-烯丙基巯基半胱氨酸对乳腺癌MCF-7(Michigan Cancer Foundation-7)细胞功能的影响。方法不同浓度SAMC对MCF-7细胞进行干预作为观察组,同时设置给予普通培养基的空白对照组,用结晶紫实验方法检测不同浓度SAMC干预下的细胞生长...目的研究S-烯丙基巯基半胱氨酸对乳腺癌MCF-7(Michigan Cancer Foundation-7)细胞功能的影响。方法不同浓度SAMC对MCF-7细胞进行干预作为观察组,同时设置给予普通培养基的空白对照组,用结晶紫实验方法检测不同浓度SAMC干预下的细胞生长速率,细胞划痕实验检测细胞的迁移能力,Transwell实验检测细胞的侵袭能力。结果与空白对照组相比,结晶紫实验结果提示SAMC可有效抑制乳腺癌MCF-7的生长速率(P<0.05),且呈现剂量依赖。细胞划痕实验检结果提示SAMC可减低MCF-7细胞的迁移能力(P<0.05),且抑制作用呈现剂量依赖。Transwell实验结果提示,100μg/mL SAMC处理后的MCF-7细胞,其细胞的侵袭能力与对照组相比未见明显变化(P>0.05),200μg/mL、300μg/mL SAMC处理后的MCF-7细胞,迁移能力显著下降(P<0.05)。结论 SAMC可抑制乳腺癌MCF-7细胞的生长、迁移及侵袭的能力,从而发挥抗乳腺癌的作用。展开更多
Objective: Our previous studies have firstly demonstrated that 17β -E2 up-regulates LRP16 gene expression in human breast cancer MCF-7 cells, and ectopic expression of the LRP16 gene promotes MCF-7 cells proliferatio...Objective: Our previous studies have firstly demonstrated that 17β -E2 up-regulates LRP16 gene expression in human breast cancer MCF-7 cells, and ectopic expression of the LRP16 gene promotes MCF-7 cells proliferation. Here, the effects of the LRP16 gene expression on growth of MCF-7 human breast cancer cells and the mechanism were further studied by establishing two stably LRP16-inhibitory MCR-7 cell lines. Methods: Hairpin small interference RNA (siRNA) strategy, by which hairpin siRNA was released by U6 promoter and was mediated by pLPC-based retroviral vector, was adopted to knockdown endogenous LRP16 level in MCF-7 cells. And the hairpin siRNA against green fluorescence protein (GFP) was used as the negative control. The suppressant efficiency of the LRP16 gene expression was confirmed by Nothern blot. Cell proliferation assay and soft agar colony formation assay were used to determine the status of the cells proliferation. Cell cycle checkpoints including cyclin E and cyclin D1 were examined by Western blot. Results: The results from cell proliferation assays suggested that down-regulation of LRP16 gene expression is capable of inhibiting MCF-7 breast cancer cell growth and down-regulation of the LRP16 gene expression is able to inhibit anchorage-independent growth of breast cancer cells in soft agar. We also demonstrated that cyclin E and cyclin D1 proteins were much lower in the LRP16-inhibitory cells than in the control cells. Conclusion: These data suggest that LRP16 gene play an important role in MCF-7 cells proliferation by regulating the pathway of the G1/S transition and may function as an important modulator in regulating the process of tumorigenesis in human breast.展开更多
基金This work was supported by NationalNatural Science Foundation of China (No. 30200095).
文摘Objective: Our previous studies have firstly demonstrated that 17β -E2 up-regulates LRP16 gene expression in human breast cancer MCF-7 cells, and ectopic expression of the LRP16 gene promotes MCF-7 cells proliferation. Here, the effects of the LRP16 gene expression on growth of MCF-7 human breast cancer cells and the mechanism were further studied by establishing two stably LRP16-inhibitory MCR-7 cell lines. Methods: Hairpin small interference RNA (siRNA) strategy, by which hairpin siRNA was released by U6 promoter and was mediated by pLPC-based retroviral vector, was adopted to knockdown endogenous LRP16 level in MCF-7 cells. And the hairpin siRNA against green fluorescence protein (GFP) was used as the negative control. The suppressant efficiency of the LRP16 gene expression was confirmed by Nothern blot. Cell proliferation assay and soft agar colony formation assay were used to determine the status of the cells proliferation. Cell cycle checkpoints including cyclin E and cyclin D1 were examined by Western blot. Results: The results from cell proliferation assays suggested that down-regulation of LRP16 gene expression is capable of inhibiting MCF-7 breast cancer cell growth and down-regulation of the LRP16 gene expression is able to inhibit anchorage-independent growth of breast cancer cells in soft agar. We also demonstrated that cyclin E and cyclin D1 proteins were much lower in the LRP16-inhibitory cells than in the control cells. Conclusion: These data suggest that LRP16 gene play an important role in MCF-7 cells proliferation by regulating the pathway of the G1/S transition and may function as an important modulator in regulating the process of tumorigenesis in human breast.
