食管鳞状细胞癌成纤维细胞hedgehog通路对SHH蛋白的应答

Fibroblasts isolated from esophageal squamous cell carcinoma response to SHH stimuli

目的探讨食管鳞状细胞癌成纤维细胞hedgehog通路是否应答SHH蛋白。方法分离、原代培养食管鳞状细胞癌成纤维细胞,用Western blot方法检测培养的细胞是否为成纤维细胞,SHH蛋白与细胞孵育后提取细胞总RNA,荧光定量PCR方法检测细胞中hedgehog通路是否应答。结果 Western blot结果显示培养细胞表达间质细胞标记Vimentin和Fibronectin,不表达上皮细胞标记E-cadherin。三株细胞中有两株应答SHH蛋白,一株不应答。结论食管鳞状细胞癌中,成纤维细胞对SHH配体是否应答存在着不同的情况,因此,检测间质中Hh通路的活性可为临床上选择对Hh抑制剂应答的肿瘤提供重要的生物标记。

Objective The aim of this study is to examine if fibroblasts isolated from esophageal squamous cell carcinoma response to SHH stimuli. Methods We isolated and cultured fibroblasts, examined their identification by westernblot. We isolated total RNA from fibroblasts treated with SHH, and tested the activity of the hedgehog pathway by real - time PCR. Results Our cultured cells expressed vimentin and fi- bronectin but not E - cadherin, indicating that they were fibroblasts. After SHH stimuli, two cells had elevated hedgehog signaling and one cell not. Conclusion Fibroblasts isolated from esophageal squamous cell carcinoma did not all response to SHH stimuli. Therefore, examining hedge- hog activity in stroma provides an important biomarker for clinical selection of tumors responded to hedgehog inhibitor.