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目的探讨三叶青乙酸乙酯提取物(ethylacetate fraction,EAF)对人胰腺癌PANC-1细胞的凋亡作用及其机制。方法 MTT法检测三叶青EAF对人胰腺癌PANC-1细胞的生长抑制作用;光镜及Hoechst 33258荧光染色分别观察药物作用后细胞形态和细胞凋亡;流式细胞仪检测细胞凋亡率;蛋白印记法(Western blot)检测凋亡蛋白Bax、Bcl-2和P53的表达情况。结果与对照组比较,三叶青EAF能显著地抑制人胰腺癌细胞PANC-1的生长,并呈明显的量-效关系。药物作用后,细胞形态改变,有明显的凋亡特征。流式细胞术检测细胞凋亡率有浓度依赖性。Western blot检测表明三叶青EAF可以促进Bax、P53蛋白表达升高,Bcl-2表达下降。结论三叶青EAF对人胰腺癌PANC-1细胞具有抑制增殖和促进凋亡作用,其机制可能与上调p53和Bax的表达,下调Bcl-2的表达有关。
Objective To investigate the apoptosis effect of ethylacetate fraction (EAF) on human pancreatic cancer PANC-1 cells and its mechanism. Methods MTT assay was used to detect the growth inhibition of human pancreatic cancer PANC-1 cells by using trichosanthin EAF. Cell morphology and apoptosis were observed by light microscopy and Hoechst 33258 staining respectively. The apoptosis rate was detected by flow cytometry. Western blot was used to detect the expression of Bax, Bcl-2 and P53 protein. Results Compared with the control group, trilobin EAF can significantly inhibit the growth of human pancreatic cancer cells PANC-1, and showed a significant dose-response relationship. After the drug effect, cell morphology changes, there are obvious characteristics of apoptosis. Flow cytometry showed a concentration-dependent rate of apoptosis. Western blot results showed that trilobal EAF can promote the expression of Bax, P53 protein, Bcl-2 expression decreased. CONCLUSIONShree Yeqing EAF can inhibit proliferation and promote apoptosis of human pancreatic cancer PANC-1 cells. The mechanism may be related to up-regulating the expression of p53 and Bax and down-regulating the expression of Bcl-2.