蝙蝠葛碱对胰腺癌BxPC-3细胞DPC4、Ki-67表达的影响

Inhibition Effect of Clerodendrum Bungei Flavonoinds on Lewis RatsLung Tumor and its Correlation to the Expression of p53,bcl-2,and bax Gene

目的观察蝙蝠葛碱（Dau）对胰腺癌细胞BxPC-3的抑制作用及其相关基因DPC4、Ki-67表达的影响。方法：取胰腺癌BxPC-3对数生长期细胞，胰酶消化后制备单细胞悬液，将细胞悬液调至1×10^5／ml浓度，接种于4个标记不同组的培养瓶中，放至培养箱中培养，培养环境为37℃、5％CO2、完全饱和湿度，24h后弃去培养基，药物干预。模型组：加正常培养基；5-Fu组：加入5-FU使其终浓度为50ug／ml；Dau低剂量组：加入Dau使其终浓度为12ug／ml；Dau中剂量组：加入Dau使其终浓度为2411g／ml；Dau高剂量组：加入Dau使其终浓度为48ug／ml；置于培养箱中培养，培养环境为37℃、5％CO2、完全饱和湿度，48h。实验采用MTT法检测Dau对人胰腺癌BxPC-3细胞生长曲线及增殖的影响以及免疫组化法检测DPC4、Ki-67蛋白的表达。结果：Dau高、中剂量组和5-Fu组对胰腺癌BxPC-3细胞的生长都有一定的抑制作用，其抑瘤率分别为67．31％、41．66％、53．06％（P〈0．01）。通过免疫组化结果显示，Dau高剂量、Dau中剂量能够提高调胰腺癌细胞BxPC-3DPC4蛋白的表达；并下调Ki-67蛋白的表达。结论：Dau中、高剂量能够有效的抑制胰腺癌细胞株Bx—PC-3的生长、增殖，并影响DPC4、Ki-67蛋白的表达。

Objective To observe the Dan on human pancreatic cancer cells BxPC -3 and effect on DPC4,Ki-67protein expression of pancreatic cancer.Methods：Pancreatic cancer BxPC-3 logarithmic growth phase cells,pancreatic enzyme preparation of single cell suspension after digestion,the cells suspension tuning to 1 ×10^5/ml concentration, vaccination in four markers in the different groups of culture bottle, put to the cultivation in the b0x,training environment for 37℃ and 5% CO2, completely saturated humidity,24 h after abandon to medium and drug intervention.Model group： normal culture medium; 5-FU group：（5-FU） makes its final concentration of 50 ug/ml;Dau：low dose group to join Dau make its final concentration of 12 ug/ml;Dau dose groups add Dau make its final concentration of 24 ug/ml;Dau：high dose group to join Dan make its final concentration of 48 ug/ml;Placed in the incubator in the training, training envi- ronment for 37× and 5%CO2,fully saturated humidity,48 h.Experiment determined by MTr method is used to detect Dau for pancreatic cancer BxPC-3 cell growth curve and the effect of proliferation and immunohistochemical method to detect the expression of DPC4 and Ki-67 protein.Results：Dan high dose and Dau mid-dose could inhibit pancreatic cancer BxPC-3 cellline.The inhibition rates were 67.31%,41.66%,53.06% respectively,which has the statistical significance（P〈0.O1）.By immunohistochemical results show, Dau can rise in pancreatic cancer cells BxPC-3 protein expression of DPC4,and cut Ki-67 expression.Conclusion：Dan high dose and Dan mid-dose can effectively inhibit the growth of pancreatic cancer cell lines BxPC-3,effect the DPC4,Ki-67 expression.