rAV-Tumstatin病毒载体构建及其生物活性检测

J4 ›› 2010, Vol. 36 ›› Issue (3): 453-455.

rAV-Tumstatin病毒载体构建及其生物活性检测

任明, 刘禄成, 魏巍, 李志, 张明, 温都苏

吉林大学第二医院泌尿外科|吉林长春 130041

收稿日期:2009-11-18 出版日期:2010-05-28 发布日期:2010-05-28
通讯作者: 任明（Tel: 0431-88796612,E-mail:AG8620@sina.com) E-mail:AG8620@sina.com
作者简介:任明（1962-）|男|吉林省农安县人|副教授|主要从事泌尿外科肿瘤治疗的研究。
基金资助:
国家自然科学基金资助课题（30972981）

Construction of rAV-Tumstatin viral vector and detection of its bioactivty

REN Meng, LIU Lu-Cheng, WEI Wei, LI Zhi, ZHANG Ming, WEN Dou-Su

Departement of Urinary Surgery,Second Hospital,Jilin University,Changchun |130041|China

Received:2009-11-18 Online:2010-05-28 Published:2010-05-28

摘要/Abstract

摘要：

目的：探讨腺病毒携带肿瘤抑素（rAV-Tumstatin）对人膀胱癌T24细胞株的影响与作用,为进一步研究其作用机制提供实验依据。方法：构建rAV-Tumstatin并感染人膀胱癌T24细胞株，将细胞分为rAV-Tumstatin组、rAV-MCS空病毒组和空白对照组，采用噻唑蓝（MTT）比色法和原位末端标记（TUNEL）法分别检测感染细胞的增殖和凋亡情况。结果： rAV-Tumstatin既可以稳定感染T24细胞也可以在该细胞内表达携带的外源基因EGFP。TUNEL法检测，rAV-Tumstatin组细胞凋亡率为32.9%，rAV-MCS空病毒组和空白对照组分别为13.5%和6.7%，3组间比较差异均有显著性（P<0.01）。MTT比色法检测T24细胞株的增殖生长能力， rAV-Tumstatin组G0/G1期60.3%，S期17.4%，G2/M期21.5%；空病毒组G0/G1期45.2%，S期29.5%，G2/M期24.8%；对照组G0/G1期43.7%，S期32.8%，G2/M期23.7%。3组间各细胞周期细胞增殖率比较差异有显著性（P<0.05）。结论： rAV-Tumstatin能抑制膀胱癌T24细胞株增殖并诱导其凋亡，对膀胱癌基因治疗的研究具有一定的价值。

关键词: 膀胱肿瘤；重组腺病毒；肿瘤抑素

Abstract:

Abstract:Objective To investigate the influence and effect of rAV-Tumstatin on T24 cell lines,and provide experimental basis　for further study on its mechanism.Methods rAV-Tumstatin　was constructed and the T24 cell lines were infected.The cells were divided into rAV-Tumstatin group,vacant viral infection group and control group.The proliferation and apoptosis of the infected cells were detected by MTT and TUNEL.Results The T24 cells could be stably infected by rAV-Tumstatin and expressed the exogenous gene EGFP. The result of TUNEL showed the apoptotic rates in rAV-Tumstatin group,vacant viral infection group and control groupwere 32.9%,13.5%,and 6.7%,respectively;there were significant differences between three groups(P<0.01).The result of MTT which detected the proliferation and growth of T24 cells respectively showed as follows: rAV-Tumstatin group,G0/G1 60.3%,S 17.4%,G2/M 21.5%;vacant viral infection group,G0/G1 45.2%,S 29.5%,G2/M 24.8%; control group,G0/G1 43.7%,S 32.8%,G2/M 23.7%;there 　were also 　significant differences between three groups(P<0.05).Conclusion rAV-Tumstatin can inhibit the proliferation and induce the apoptosis of T24 cells,and it is very valuable for the gene therapy of bladder cancer.

Key words: bladder neoplasms;recombinant adenovirus;Tumstatin

中图分类号:

R735.14

任明, 刘禄成, 魏巍, 李志, 张明, 温都苏. rAV-Tumstatin病毒载体构建及其生物活性检测[J]. J4, 2010, 36(3): 453-455.

REN Meng, LIU Lu-Cheng, WEI Wei, LI Zhi, ZHANG Ming, WEN Dou-Su. Construction of rAV-Tumstatin viral vector and detection of its bioactivty[J]. J4, 2010, 36(3): 453-455.