山慈菇提取物对人结直肠癌SW480细胞增殖和凋亡的影响

Effects of Iphigenia Indica Extract on Proliferation and Apoptosis of Human Colorectal Cancer Cell Line SW480

目的:观察不同浓度山慈菇提取物对人结直肠癌SW480细胞增殖和凋亡的影响,并初步探讨其作用机制。方法:以SW480细胞为研究对象,设立不同浓度山慈菇提取物与空白组,采用CCK-8法检测SW480细胞的活性,克隆形成实验检测SW480细胞克隆形成能力,Hoechst 33258染色检测SW480细胞凋亡时细胞核的形态学变化,Western blot法检测SW480细胞中星形细胞上调基因-1(Astrocyte elevated gene-1,AEG-1)、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)和Bcl-2-associated X的蛋白质(Bax)表达水平,最后采用shRNA干扰技术沉默SW480细胞中AEG-1基因的表达检测AEG-1基因是否能影响SW480细胞的增殖以及Bcl-2和Bax的蛋白表达。结果:与空白组比较,山慈菇提取物能显著抑制SW480细胞活性和细胞克隆形成能力(P<0.05);Hoechst 33258染色后观察可见细胞形态皱缩并形成凋亡小体;与空白组比较,山慈菇提取物组AEG-1和Bcl-2蛋白表达水平明显降低,Bax蛋白表达水平则明显增加(P<0.05);经shRNA沉默AEG-1基因表达后,SW480细胞中AEG-1蛋白水平显著下降(P<0.05);与阴性对照组比较,AEG-1沉默慢病毒转染组SW480细胞中Bcl-2蛋白表达明显降低(P<0.05),Bax蛋白表达明显增加(P<0.05),细胞活性和细胞克隆形成能力均受到明显抑制(P<0.05)。结论:山慈菇提取物具有明显抑制SW480细胞增殖并诱导凋亡的作用,其机制可能与抑制AEG-1蛋白表达、继而下调Bcl-2蛋白的表达水平、上调Bax蛋白表达水平有关。

Objective:To observe the effects of Iphigenia Indica extract on the proliferation and apoptosis of colorectal cancer cell carcinoma SW480 cells,and to explore its mechanism.Methods:SW480 cells were treated with different concentrations of Iphigenia indica extract.The activity of SW480 cells was detected by CCK8 method;the colony forming ability of SW480 cells was detected by clone formation experiment.The morphological changes of SW480 cells were detected by Hoechst 33258 staining,and the expression levels of AEG-1,Bcl-2 and Bax were detected by Western blot.Finally,shRNA was used to silence the expression of AEG-1 gene in SW480 cells to detect whether AEG-1 gene can affect the proliferation of SW480 cells and the protein expression of Bcl-2 and Bax.Results:Compared with the blank group,the application of the Iphigenia indica extract could significantly inhibit the activity of SW480 cells and decrease the ability of cell clone formation(P<0.05);after Hoechst 33258 staining,the cell morphology was observed to shrink and apoptotic bodies were formed;compared with the blank group,the expression level of AEG-1 and Bcl-2 protein was significantly decreased(P<0.05),and the expression level of Bax protein was significantly increased(P<0.05).After shRNA silencing the expression of AEG-1 gene,the expression level of AEG-1 protein in SW480 cells decreased significantly(P<0.05);compared with the negative control group,the expression of Bcl-2 protein in SW480 cells in the AEG-1 silencing group decreased significantly(P<0.05),the expression of Bax protein increased significantly(P<0.05),the cell activity and the ability of cell clone formation were inhibited significantly(P<0.05).Conclusion:Iphigenia indica extract can obviously inhibit the proliferation and induce apoptosis of SW480 cells.Its mechanism may be related to the inhibition of AEG-1 protein expression,then down-regulation of Bcl-2 protein expression and up-regulation of Bax protein expression.