贵州医科大学学报

2019, v.44;No.229(10) 1173-1178

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吉西他滨联合厄洛替尼通过抑制突变型p53蛋白表达促进胰腺癌细胞凋亡
Gemcitabine Combined with Erlotinib Promotes Apoptosis of Pancreatic Cancer Cells by Inhibiting the Expression of Mutant p53

黄欣昊;柳千帆;宋春灼;朱海涛;
HUANG Xinhao;LIU Qianfan;SONG Chunzuo;ZHU Haitao;Department of Clinical Medicine, Guizhou Medical University;Clinical Research Center of the Affiliated Hospital of Guizhou Medical University;Department of Hepatobiliary Surgery, The Affiliated Hospital of Guizhou Medical University;

摘要(Abstract):

目的:探讨吉西他滨(GEM)联合厄洛替尼(ERL)通过抑制突变型p53蛋白表达促进胰腺癌细胞凋亡的分子机制。方法:选用人胰腺癌细胞株PANC-1对数生长期细胞,将细胞分为对照组(无处理)、ERL组(ERL干预浓度12 nmol/L)、GEM组(GEM干预浓度80 nmol/L)、ERL联合GEM组(联合组,GEM干预浓度为4 nmol/L,ERL干预浓度为20 nmol/L),采用Annexin-V APC/7AAD法检测各组细胞的凋亡,采用实时荧光定量PCR检测各组细胞p53基因表达,采用Western blot法检测各组细胞p53蛋白和B淋巴细胞瘤-2相关蛋白X(BAX)的表达量。结果:联合组PANC-1细胞的凋亡率高于对照组、GEM组及ERL组(P<0.05或P<0.01);与对照组比较,GEM组、ERL组和联合组PANC-1细胞中p53基因的表达量均明显下降(P<0.001);联合组PANC-1细胞中p53蛋白表达量分别低于对照组、GEM组和ERL组(P<0.05或P<0.01),但BAX蛋白表达量分别高于对照组、GEM组和ERL组(P<0.05或P<0.01)。结论:GEM联合ERL可能下调突变型p53,诱导并上调BAX的表达,从而发挥促进肿瘤凋亡的作用。
Objective: To investigate the effect of Gemcitabine(GEM) combined with Erlotinib(ERL) on the apoptosis of human pancreatic cancer cells and its possible molecular mechanism. Methods: Human pancreatic cancer cells in logarithmic growth phase(PANC-1) were selected and divided into the control group(without treatment), ERL group(ERL intervention concentration 12 nmol/L), and GEM group(GEM intervention concentration 80 nmol/L), GEM and ERL group(combined group with 4 nmol/L GEM intervention concentration,, and 20 nmol/L ERL intervention concentration). All the cells from each treatment group were cultured for 24 hours and collected. Apoptosis of cells was examined by Annexin-V APC/7 AAD method. The expression of p53 gene in the cells of each treatment group was detected by real-time fluorescent quantitative PCR. The expressions of p53 and BAX of the cells of all the groups were detected by Western Blot, and compared with those of total proteins. Results: The apoptotic rate of PANC-1 in the pancreatic cancer cells of the combined group was higher than that in the control group, ERL group and GEM group(P<0.05). The expression of p53 gene in PANC-1 cells decreased more obviously in ERL group, GEM group and the combined group than in the control group(P<0.05). The expression of p53 decreased and its expression of BAX increased in the combined group more obviously than in the control, ERL and GEM groups(P<0.05). At the same time, the expression of p53 in pancreatic cancer cell line PANC-1 in the combined group was lower than that in ERL and GEM groups(P<0.05), and its expression of BAX was higher than that ERL and GEM groups(P<0.05). Conclusion: The therapy of GEM combined with ERL may play a better role in promoting tumor apoptosis by inducing p53 signaling pathway, down-regulating mutant p53, and inducing and up-regulating the expression of BAX.

关键词(KeyWords): 胰腺肿瘤;细胞凋亡;厄洛替尼;吉西他滨;联合用药;p53蛋白
pancreatic cancer;apoptosis;erlotinib;gemcitabine;combination therapy;p53 protein

Abstract:

Keywords:

基金项目(Foundation): 贵州医科大学2018年度学术新苗培养及创新探索专项项目[黔科合平台人才(2018)5779-31]

作者(Authors): 黄欣昊;柳千帆;宋春灼;朱海涛;
HUANG Xinhao;LIU Qianfan;SONG Chunzuo;ZHU Haitao;Department of Clinical Medicine, Guizhou Medical University;Clinical Research Center of the Affiliated Hospital of Guizhou Medical University;Department of Hepatobiliary Surgery, The Affiliated Hospital of Guizhou Medical University;

DOI: 10.19367/j.cnki.1000-2707.2019.10.011

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