Adeno-associated virus 2-mediated intratumoral prostate cancer gene therapy: Long-term maspin expression efficiently suppresses tumor growth
Adeno-associated virus 2-mediated intratumoral prostate cancer gene therapy: Long-term maspin expression efficiently suppresses tumor growth
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DOI：
10.1089/hum.2005.16.699
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发表时间：
2005-06-01
影响因子：
4.2
通讯作者：
Kumon, H
中科院分区：
文献类型：
--
作者：
Watanabe, M;Nasu, Y;Kumon, H
Maspin是丝氨酸蛋白酶抑制剂的成员，并且Maspin基因是一种肿瘤抑制基因，在大部分前列腺癌中下调。我们评估了使用腺相关病毒（AAV，血清型2）载体编码maspin作为体内基因治疗人前列腺癌的手段。裸鼠皮下形成的LNCaP或DU 145肿瘤的TUNEL测定显示，与AAV-LacZ治疗相比，瘤内AAV介导的maspin表达显著上调凋亡细胞的数量。免疫荧光双染色显示，与AAV-GFP介导的GFP表达细胞相比，AAV-maspin介导的maspin表达细胞中凋亡细胞的百分比显著增加。此外，与对照肿瘤相比，在AAV-maspin处理的肿瘤中观察到显著更少的CD 31阳性微血管。这些治疗反应与肿瘤中持续的maspin表达高度相关，通过蛋白质印迹分析证实，直到治疗后至少第56天。最后，瘤内递送AAV-maspin显著抑制LNCaP和DU 145肿瘤的生长并改善小鼠的存活。我们的结论是，AAV介导的maspin表达延长有效地抑制人前列腺肿瘤的生长在体内诱导细胞凋亡和抑制血管生成。
Maspin is a member of the serine protease inhibitors and the maspin gene, a tumor suppressor gene, is down-regulated in a large fraction of prostate cancers. We evaluated the use of adeno-associated virus (AAV, serotype 2) vector encoding maspin as a means for in vivo gene therapy for human prostate cancer. TUNEL assay of subcutaneously formed LNCaP or DU145 tumors in nude mice showed that intratumoral AAV-mediated maspin expression significantly upregulated the number of apoptotic cells compared with AAV-LacZ treatment. Immunofluorescence double staining for maspin protein and apoptosis in LNCaP tumors showed that the percentage of apoptotic cells in AAV-maspin-mediated maspin-expressing cells was significantly high compared with that in AAV-GFP-mediated GFP-expressing cells. Moreover, significantly fewer CD31-positive microvessels were observed in AAV-maspin-treated tumors compared with the control tumors. These therapeutic responses were highly correlated to persistent maspin expression in tumors, confirmed by Western blot analysis until at least day 56 after treatment. Finally, intratumoral delivery of AAV-maspin significantly suppressed growth of LNCaP and DU145 tumors and improved survival of mice. We conclude that AAV-mediated prolonged maspin expression efficiently suppresses human prostate tumor growth in vivo by apoptosis induction and inhibition of angiogenesis.