Preclinical study for gene therapy against bladder cancer utilizing gelsolin gene and a dominant negative ras mutant

利用凝溶胶蛋白基因和显性失活ras突变体治疗膀胱癌的临床前研究

• 批准号: 10470327
• 负责人: SHINOHARA Nobuo
• 金额: $ 3.46万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10470327/
• 关键词: Human bladder cancer cell lines; ras suppressor mutant; Gene Transfer; Growth suppression; gelsolin; intravesical administration; Ras抑制変異体; アデノウィルスベクター; 細胞増殖抑制; 膀胱癌

We investigated the in vivo efficacy of gene therapy with the gelsolin gene in nude mice with human bladder cancer cell lines. This form of gene therapy, in which retrovirus producer cells were introduced, resulted in marked and reproducible tumor growth inbibition and prolonged survival time in the majority of animals tested.We also examined whether N116Y, which derived from the v-H-ras oncogene by substituting the asparagine-116 with tyrosine, can also inhibit the growth of human bladder cancer cell lines. Initially, we examined suppressive effects of the N116Y on transformed phenotypes and Ras signaling pathways. These studies demonstrated that the N116Y was capable of suppressing the transformed phenotype in human bladder cancer cell line, even when its expression is relative low. To analyze the mechanism of suppressive effects of N116Y, we examined the expression of Grb2 and Sos proteins, which are probably affected by N116Y, in human bladder cancer cell lines. These results showed that expression of Grb2 and Sos proteins were significantly increased in human bladder cancer cell lines in comparison with cultured normal urothelial cells. Lastly, we examined the inhibitory effects of AdCMV-N116Y on orthotopically-implanted human bladder cancer cells in nude mice. The in vivo growth of implanted bladder cancer was significantly inhibited by transurethral inoculation of AdCMV-N116Y.These results suggest that the inoculation of AdCMV-N116Y into the bladder is highly effective for remission of bladder cancer, and this therapy would be very useful as a novel treatment of human bladder cancer.

我们研究了明胶蛋白基因对人膀胱癌裸鼠移植瘤的体内治疗效果。这种引入逆转录病毒产生细胞的基因治疗，在大多数受试动物中导致了显著的和可重复性的肿瘤生长抑制，并延长了生存时间。我们还检测了由v-H-ras癌基因衍生的N116Y是否也能抑制人膀胱癌细胞系的生长。最初，我们研究了N116Y对转化表型和RAS信号通路的抑制作用。这些研究表明，N116Y能够抑制人膀胱癌细胞系的转化表型，即使其表达水平相对较低。为了分析N116Y的抑制作用机制，我们检测了可能受N116Y影响的Grb2和SOS蛋白在人膀胱癌细胞系中的表达。这些结果表明，与培养的正常尿路上皮细胞相比，人膀胱癌细胞系中Grb2和SOS蛋白的表达显著增加。最后，我们检测了AdCMV-N116Y对人膀胱癌裸鼠原位移植瘤的抑制作用。经尿道接种AdCMV-N116Y可显著抑制膀胱癌的体内生长，提示膀胱内接种AdCMV-N116Y可有效缓解膀胱癌，有望成为治疗膀胱癌的新方法。

项目成果

Tanaka M.et al.: "Gelsolin gene therapy by retrovirus producer cells for human bladder cancer in nude mice"Cancer Gene Therapy. 6. 482-487 (1999)

Tanaka M.等人：“通过逆转录病毒产生细胞对裸鼠中的人膀胱癌进行凝溶胶蛋白基因治疗”癌症基因治疗。

Watanabe T.et al.: "Suppressive effects dominant negative ras mutant N116Y on transformed phenotypes of human bladder cancer cells"Cancer Letters. (in press).

Watanabe T.等人：“显性失活 ras 突变体 N116Y 对人类膀胱癌细胞转化表型的抑制作用”《癌症快报》。

Watanabe, T., et al.: "Suppressive effects of dominant negative ras mutant N116Y on tansformed phenotypes of human bladder cancer cells"Cancer Letters. (in press).

Watanabe, T. 等人：“显性失活 ras 突变体 N116Y 对人类膀胱癌细胞转化表型的抑制作用”Cancer Letters。

Tanaka M et al.: "Gene therapy for bladder cancer"Cancer Therapy & Host (Japanese). 10(1). 43-49 (1998)

Tanaka M 等人：“膀胱癌的基因治疗”癌症治疗

Shinohara N.et al.: "The significance of C3G, one of ras guanine nucleotide exchange factors, in the proliferation of human bladder cancer cell lines"J.Urol.. 159. 1060A (1998)

Shinohara N.等人：“C3G（一种 ras 鸟嘌呤核苷酸交换因子）在人膀胱癌细胞系增殖中的意义”J.Urol.. 159. 1060A (1998)