血管紧张素转换酶2对人胰腺癌细胞BxPC3体外增殖及成瘤性的效应研究  

Effects of angiotensin-converting enzyme 2 on proliferation and tumorigenicity of human pancreatic cancer cell line BxPC3

作　　者：周琳[1] 彭孝倩 刘源[1] 史成章[1] 张连峰[1] ZHOU Lin;PENG Xiaoqian;LIU Yuan;SHI Chengzhang;ZHANG Lianfeng(Department of Gastroenterology,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院消化科,河南郑州450052

出　　处：《胃肠病学和肝病学杂志》2020年第8期926-929,共4页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家自然科学基金资助项目(81472325)。

摘　　要：目的探讨血管紧张素转换酶2(ACE2)基因对人胰腺癌细胞BxPC3在体外的增殖及成瘤性的影响及其初步机制。方法通过脂质体转染法建立稳定过表达ACE2基因的胰腺癌BxPC3细胞株。设实验组(转染ACE2质粒组,BxPC3/ACE2)、阴性对照组(转染GFP对照质粒组,BxPC3/GFP)及空白对照组(未转染组,BxPC3),Western blotting法验证转染后各组细胞ACE2蛋白的表达;MTT法检测细胞增殖变化,克隆形成实验检测细胞在体外的成瘤性;流式细胞仪和Caspase-3蛋白检测观察各组细胞凋亡情况。结果 ACE2表达质粒转染胰腺癌细胞BxPC3后,BxPC3中ACE2蛋白表达明显上调。在48 h、72 h时间点,BxPC3/ACE2细胞的增殖能力下降,与阴性对照组及空白对照组相比,差异有统计学意义(P<0.05)。克隆形成实验显示,BxPC3/ACE2组的细胞克隆与两对照组相比,不仅数目少(P<0.05),且克隆体积也相对较小。流式细胞计数检测,从撤除血清后的24 h开始,实验组细胞凋亡率明显增加,与两对照组相比差异有统计学意义(P<0.05);Caspase-3蛋白印迹发现典型凋亡特征片段。结论 ACE2基因可通过促进胰腺癌细胞BxPC3的凋亡,进而抑制其在体外的增殖及成瘤能力。Objective To investigate the effects and mechanisms of angiotensin-converting enzyme 2(ACE2) overexpression on the proliferaion and tumorigenicity of human pancreatic cancer cell line BxPC3 in vitro. Methods ACE2 expression vector was constructed and transfected into pancreatic cancer cell line BxPC3. Cells were divided into ACE2 group(BxPC3/ACE2, treated with ACE2 expression vector), negative control group(BxPC3/GFP, treated with GFP expression vector) and untreated group(BxPC3). Western blotting was used to detect the ACE2 and Caspase-3 proteins level of three groups. The cell growth was detected by MTT. Colony formation assay was used to detect the tumorigenicity of human pancreatic cancer cells. The apoptosis of pancreatic cancer cells was measured by flow cytometry and Caspase-3 protein detection. Results Proliferation rate of BxPC3/ACE2 cells was remarkably lower than BxPC3 and BxPC3/GFP cells(P<0.05). Compared with their controls, cells transfected with ACE2 exhibited not only a less amount but also a smaller size of colonies. The cell apoptic rate in BxPC3/ACE2 cells was higher in BxPC3/ACE2 group than that in negative control and untreated groups(P<0.05). Conclusion Stable overexpression of ACE2 inhibited the proliferation and tumorigenicity of BxPC3 cell iv vitro. These results provide evidence that ACE2 plays a pivotal role in the development of pancreatic cancer.

关 键 词：胰腺癌 血管紧张素转换酶2 增殖 凋亡 

分 类 号：R735.9[医药卫生—肿瘤]