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目的:探讨自噬抑制剂3-甲基腺嘌呤(3-MA)对人大肠癌SW480细胞生长与Notch1蛋白表达的影响。方法:将3-MA(5 mmol/L)作用于SW480细胞24 h后(以培养相同时间无处理的SW480细胞为对照),分别免疫组化和Western blot法检测细胞Notch1蛋白的表达,用CCK-8法和Annexin/PI双染法检测细胞增殖与凋亡。结果:免疫组化与Western blot结果均显示,3-MA作用后,SW480细胞Notch1蛋白的表达明显下调(均P<0.05);增殖与凋亡检测结果显示,3-MA作用后,SW480细胞增殖率明显降低,而凋亡率明显增加(均P<0.05)。结论:3-MA能抑制结直肠癌细胞的增殖并促进其凋亡,该作用可能与3-MA抑制Notch1蛋白表达从而改变细胞自噬水平有关。
Objective: To investigate the effect of 3-methyladenine (3-MA), an inhibitor of autophagy, on the growth and the expression of Notch1 protein in human colorectal cancer cell line SW480. Methods: SW480 cells were treated with 3-MA (5 mmol / L) for 24 h (control SW480 cells untreated for the same time). The expression of Notch1 protein was detected by immunohistochemistry and Western blot respectively. The expression of Notch1 protein was detected by CCK -8 method and Annexin / PI double staining method to detect cell proliferation and apoptosis. Results: The results of immunohistochemistry and Western blot showed that the expression of Notch1 protein in SW480 cells was significantly down-regulated after treated with 3-MA (all P <0.05). The results of proliferation and apoptosis assay showed that SW480 cells proliferated The rate of apoptosis was significantly decreased (P <0.05). Conclusion: 3-MA can inhibit the proliferation and promote the apoptosis of colorectal cancer cells, which may be related to the inhibition of Notch1 protein expression and the change of autophagy by 3-MA.