hCAP-18/LL-37基因转染对Lewis肺癌细胞增殖及凋亡的影响

J4 ›› 2009, Vol. 35 ›› Issue (6): 1040-1043.

hCAP-18/LL-37基因转染对Lewis肺癌细胞增殖及凋亡的影响

许娜^1,2|袁红艳¹|王凤丽¹|廖翔宇¹|刘超¹|常雅萍¹

1．吉林大学基础医学院免疫学教研室|吉林长春 130021；2．吉林医药学院教务处|吉林吉林 132013

收稿日期:2009-05-08 出版日期:2009-11-28 发布日期:2009-11-28
作者简介:许娜（1977-）|女|吉林省吉林市人|讲师|在读医学博士|主要从事肿瘤免疫学研究。
基金资助:
吉林省科技厅科技发展计划项目资助课题(200705357)

Effect of hCAP-18/LL-37 gene transfection on proliferation and apoptosis of Lewis cells

XU Na ^1,2,YUAN Hong-yan ¹,WANG Feng-li ¹,LIAO Xiang-yu ¹,LIU Chao ¹,CHANG Ya-ping¹

1.Department of Immunology,School of Basic Medical Sciences|Jilin University|Changchun 130021|China；2. Dean’s Office,Jilin Medical College,Jilin 132013|China

Received:2009-05-08 Online:2009-11-28 Published:2009-11-28
Contact: 常雅萍 E-mail:cyping823@163.com

摘要/Abstract

摘要：

目的:观察人源抗菌肽hCAP-18/LL-37基因转染对Lewis肺癌细胞增殖的抑制作用，探讨其抗肿瘤的可能机制。方法:将含hCAP-18/LL-37的真核表达质粒瞬时转染Lewis肺癌细胞,同时设正常对照组、转染空载体组和转染重组基因组，MTT比色检测Lewis肺癌细胞转染后48和72 h的增殖活性，流式细胞术和荧光染色检测细胞凋亡。免疫细胞化学染色检测Bax、Bcl-2蛋白表达。结果：转染重组基因组Lewis细胞增殖受到明显抑制，与转染空载体组比较，转染后48和72 h的抑制率分别为3.51%和17.65%；流式细胞术检测转染重组基因组凋亡率为7.4%，正常对照组凋亡率为4.4%，转染空载体组凋亡率为5.2%；免疫细胞化学染色，转染重
组基因组Lewis细胞Bax蛋白表达增加，与对照组比较差异有显著性(P<0.05)；Bcl-2蛋白表达减少，但与对照组比较差异无显著性(P<0.05)。结论：hCAP-18/LL-37基因瞬时转染Le
wis肺癌细胞，可抑制肿瘤细胞生长、调控凋亡相关基因Bax、 Bcl-2表达，诱导肿瘤细胞凋亡。

关键词: hCAP-18/LL-37；瞬时转染；癌, Lewis肺；细胞凋亡

Abstract:

Objective To investigate the inhibitory effect of hCAP-18/LL-37 gene transfection on proliferation in Lewis cells,and to study the possible antitumor mechanism.Methods DNA of hCAP-18/LL-37 transient transfection mediated by Polyfect was employed.The experimental cells were classified into untransfected cells,pcDNA4/Myc-His transfected cells and pcDNA4/Myc-His- LL-37 transfected cells.The proliferating abilities of Lewis cells 48 and 72h after transfection were measured by MTT colorimetric assay.The apoptosis of transfected Lewis cells was detected by flow cytometry and fluorescence microscope.The expressions of Bax and Bcl-2 in transfected Lewis cells were determined by cellular and immunohistochemical technique.Results The proliferating viability of Lewis cells transfected by the recombinant plasmid was significantly inhibited,the inhibitory rate of growth was 3.51% and 17.65% compared with pcDNA4/Myc-His transfected cells.The apoptotic rate were 7.4%,4.4%,5.2% in untransfected cells,pcDNA4/Myc-His cells and pcDNA4/Myc-His- LL-37 cells, respectivly, by FCM measurement.The protein expression of Bax in transfected Lewis cells was increased,and there was significant difference compared with control(P< 0.05).The protein expression of Bcl-2 in transfected Lewis cells was decreased,but there was no significant difference compared with control(P>0.05).
Conclusion hCAP-18/LL-37 gene transfection can induce apoptosis of Lewis cells by up-regulation of Bax and down-regulation of Bcl-2 and inhibit the proliferating ability of Lewis cells.

Key words: hCAP-18/LL-37;transient transfection;carcinoma,Lewis lung;apoptosis

中图分类号:

许娜, 袁红艳, 王凤丽, 廖翔宇, 刘超, 常雅萍. hCAP-18/LL-37基因转染对Lewis肺癌细胞增殖及凋亡的影响[J]. J4, 2009, 35(6): 1040-1043.

HU Na, YUAN Gong-Yan, WANG Feng-Li, LIAO Xiang-Yu, LIU Chao, CHANG Ya-Ping. Effect of hCAP-18/LL-37 gene transfection on proliferation and apoptosis of Lewis cells[J]. J4, 2009, 35(6): 1040-1043.