Nakamura Masafumi, Kubo Makoto, Yanai Kosuke, Mikami Yoshiko, Ikebe Mio, Nagai Shuntaro, Yamaguchi Koji, Tanaka Masao, Katano Mitsuo
Department of Cancer Therapy and Research, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
Anticancer Res. 2007 Nov-Dec;27(6A):3743-7.
The hedgehog (Hh) signaling pathway is aberrantly activated in many human carcinomas including pancreatic cancer and regulates tumor cell growth. Overproduction of sonic hedgehog (Shh), a ligand of the Hh signaling pathway, increases the Hh signaling activity through transmitting the signal to patched-1 (Ptch1), the receptor of the Hh signaling pathway.
a-Ptch1 antibodies were raised against an oligo-peptide, designed according to the Ptch1 aminoacid sequence. The specificity of a-Ptch1 was examined by immunoblotting and immuno-fluorescence, and biological effects were detected by RT-PCR and cell proliferation assay using two pancreatic cancer cell lines, Panc1 and SUIT-2.
a-Ptch1 recognized a 160 kDa protein as shown by immunoblotting and cell surface staining of pancreatic cancer cells. Incubation with a-Ptch1 suppressed Hh signaling activity and proliferation of pancreatic cancer cells.
These results provide a new strategy for controling Hh dependent development of pancreatic cancer and other Hh related carcinomas.
刺猬信号通路(Hh)在包括胰腺癌在内的多种人类癌症中异常激活,并调节肿瘤细胞生长。Hh信号通路的配体音猬因子(Shh)的过量产生通过将信号传递给Hh信号通路的受体 patched-1(Ptch1)来增加Hh信号活性。
根据Ptch1氨基酸序列设计合成寡肽,以此制备抗Ptch1抗体(α-Ptch1)。通过免疫印迹和免疫荧光检测α-Ptch1的特异性,并使用两种胰腺癌细胞系Panc1和SUIT-2,通过逆转录聚合酶链反应(RT-PCR)和细胞增殖试验检测其生物学效应。
免疫印迹和胰腺癌细胞表面染色显示,α-Ptch1识别出一种160 kDa的蛋白质。用α-Ptch1处理可抑制胰腺癌细胞的Hh信号活性和增殖。
这些结果为控制胰腺癌和其他Hh相关癌症的Hh依赖性发展提供了新策略。