摘要
目的：探讨小鼠β-防御素-2（beta defensin 2，BD2）对 SiHa 细胞生长的抑制作用及其机制。方法将pcDNA3．1（＋）/mBD 2、pcDNA3．1（＋）/rmBD 2质粒转染 SiHa 获得的稳定转染细胞 SiHa/M、SiHa/R 裂解，用细胞裂解上清进行 Western blot 检测 mBD2蛋白表达；同时采用细胞活力分析对转染细胞的细胞活力进行测定，采用 AnnexinⅤ、PI 流式细胞分析法和 TUNEL 法检测细胞凋亡情况，并计算细胞凋亡指数。结果 SiHa/M、SiHa/R 组在4～6 Ku 区域出现目标条带；SiHa/M、SiHa/R 组细胞在贴壁后2～3 d 细胞活性明显低于 SiHa 细胞（P ＝0．018），但随着时间推移，细胞活性开始下降，3组细胞的细胞活力差异无统计学意义（P ＝0．374）；SiHa/M、SiHa/R 及 SiHa 组 Annexin Ⅴ＋/PI＋双阳性细胞的百分率分别为2．4％、2．3％、2．3％；Annexin Ⅴ＋/PI-细胞分别为0．9％、1．2％、1．5％，3组差异无统计学意义（P ＝0．743）；SiHa/M、SiHa/R 组凋亡细胞发生率较高，分别达到40．7％、30．2％，而 SiHa 组细胞凋亡率只有7．3％，SiHa/M、SiHa/R 组细胞凋亡率明显高于 SiHa 组，差异有统计学意义（P ＝0．004）；SiHa/M 组凋亡发生率高于 SiHa/R 组，差异有统计学意义（P ＝0．003）。结论 mBD 2能够抑制 SiHa 细胞的生长和诱导细胞凋亡。
Objective In this study,we hope to discover possible anti-tumor mechanism by observed the in-hibitory effects of mBD2 on uterine cervix cancer in vitro.Methods The pcDNA3.1（＋）/mBD2,pcDNA3. 1（＋）/rmBD2 and pcDNA3.1（＋）were transformed into the SiHa,an uterine cervix cancer cell line.The expression of target proteins were detected with Western Blot.Cell Viability Analyzer,TUNEL and An-nexin Ⅴ were applied to detect effects of mBD2 on SiHa appotosis and viability.Results The recombinant eukaryotic expression plasmid were transfected into SiHa cells.The target protein was confirmed correctly by western Blot.The viabilitys of SiHa/M,SiHa/R showed significant difference with SiHa （P =0.018））. The Annexin Ⅴ ＋/PI＋ rate of SiHa/M,SiHa/R and SiHa were 2.4%,2.3%,2.3%,and Annexin Ⅴ ＋/PI- were 0.9%,1.2%,1.5%.But there were not statistical difference in three group （P =0.743）.The ap-optotic indexs of SiHa/M,SiHa/R were 40.7%,30.2%,while SiHa was 7.3% （P =0.004）.Conclusion mBD2 could inhibit SiHa cell growth,and induce its apoptosis.
出处
《新疆医科大学学报》
CAS
2014年第3期287-291,共5页
Journal of Xinjiang Medical University
基金
新疆维吾尔自治区自然科学基金(2011211A048)
新疆维吾尔自治区高校重点基金(XJEDU2009120)
关键词
Β防御素
细胞活力
流式细胞术
凋亡指数
TUNEL
beta defensin
cytoactived
flow cytometry
TUNEL
apoptosis index
