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
This study was conducted to identify a cheap and suitable culture medium for the mycelial growth and sporulation of Sternphylium vesicarium and to determine the cultural and morphological variability of this pathogen....This study was conducted to identify a cheap and suitable culture medium for the mycelial growth and sporulation of Sternphylium vesicarium and to determine the cultural and morphological variability of this pathogen. A total of 24 isolates of S. vesicarium collected from eight different onion growing areas were characterized in terms of cultural and morphological aspects. Front colony colors were greenish brown to dirty white, deep grey to whitish, light grey to whitish, deep greenish white, light grey and dirty white to greenish. Reverse colony colors were brown, deep brown and light brown. Colony shapes were circular and irregular with umbonate, raised and flat type colony elevation. Colony textures were cottony, fluffy and velvety with entire, undulate and filiform type colony margin. Among the culture media, V-7 juice agar found to be the most suitable culture media for mycclial growth of S. vesicarium. The sporulation of the fungus was remarkably influenced by V-7 juice mixed with potato dextrose agar （PDA） media, this media exhibited the highest sporulation （87.76-169.0/mm^2） of S. vesicariumin comparison with other media. The minimum days （28 d to 31 d） for conidial production were observed on V-7 juice agar medium. The length of conidia varied from 14.6 μm to 30.6 μm. The maximum mean length of conidia was 29.97 μm found in isolate DSSA, while the minimum mean length 17.36 μm was found in isolate MSMM 02. The breadth of conidia ranged from 4.7 μm to 15.7μm. The maximum mean breadth of conidia was 12.55 μm found in the isolate DSSA, while the minimum mean breadth 9.760 μm was found in the isolate CCKH 02. The horizontal septation varied from l to 3 and the longitudinal septation varied from 0 to 4.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.
文摘This study was conducted to identify a cheap and suitable culture medium for the mycelial growth and sporulation of Sternphylium vesicarium and to determine the cultural and morphological variability of this pathogen. A total of 24 isolates of S. vesicarium collected from eight different onion growing areas were characterized in terms of cultural and morphological aspects. Front colony colors were greenish brown to dirty white, deep grey to whitish, light grey to whitish, deep greenish white, light grey and dirty white to greenish. Reverse colony colors were brown, deep brown and light brown. Colony shapes were circular and irregular with umbonate, raised and flat type colony elevation. Colony textures were cottony, fluffy and velvety with entire, undulate and filiform type colony margin. Among the culture media, V-7 juice agar found to be the most suitable culture media for mycclial growth of S. vesicarium. The sporulation of the fungus was remarkably influenced by V-7 juice mixed with potato dextrose agar （PDA） media, this media exhibited the highest sporulation （87.76-169.0/mm^2） of S. vesicariumin comparison with other media. The minimum days （28 d to 31 d） for conidial production were observed on V-7 juice agar medium. The length of conidia varied from 14.6 μm to 30.6 μm. The maximum mean length of conidia was 29.97 μm found in isolate DSSA, while the minimum mean length 17.36 μm was found in isolate MSMM 02. The breadth of conidia ranged from 4.7 μm to 15.7μm. The maximum mean breadth of conidia was 12.55 μm found in the isolate DSSA, while the minimum mean breadth 9.760 μm was found in the isolate CCKH 02. The horizontal septation varied from l to 3 and the longitudinal septation varied from 0 to 4.
