The purpose of this study was to evaluate the expression of Hedgehog (HH) signaling molecules (SHH and GLI-1) by cancer-associated fibroblasts (CAF) in oral squamous cell carcinoma (OSCC). Immunohistochemistry was used to detect molecular HH signaling and CAF-related protein expression, including α-SMA and S100A4, in 70 samples of human OSCC. The colocalization of α-SMA and S100A4 with SHH was also evaluated by double-staining. In vitro study was performed using primary normal oral fibroblast (NOF) and CAF through immunofluorescence and Western Blot for CAF-proteins, SHH, and GLI-1. Forty-five cases (64.28%) were positive for α-SMA exclusively in tumor stroma, and S100A4 was identified in the cytoplasm of CAFs in 94.28% (n = 66) of the cases. With respect to stromal cells, 64 (91.43%) OSCC cases were positive for SHH, and 31 were positive for GLI-1 (44.29%); positive correlations were found between SHH and α-SMA (p < 0.0001, φ = 0.51), as well as between SHH and S100A4 (p = 0.087, φ = 0.94). Protein expression of SHH and GLI-1 was observed in primary CAFs and NOFs. Although SHH was found to be localized in the cellular cytoplasm of both cell types, GLI-1 was present only in the nuclei of CAF. Our results indicate that CAFs are not only potential sources of HH ligands in tumor stroma, but may also respond to HH signaling through nuclear GLI-1 activation. We further observed that elevated SHH expression by OSCC cells was associated with higher CAF density, reinforcing the chemoattractant role played by these molecules.

这项研究的目的是评估在口腔鳞状细胞癌（OSCC）中癌相关成纤维细胞（CAF）对刺猬（HH）信号分子（SHH和GLI-1）的表达。免疫组织化学被用来检测人OSCC的70个样品中的分子HH信号和CAF相关蛋白表达，包括α-SMA和S100A4。还通过双重染色评估了α-SMA和S100A4与SHH的共定位。使用原发性正常口腔成纤维细胞（NOF）和CAF，通过免疫荧光和Western Blot对CAF蛋白，SHH和GLI-1进行体外研究。仅在肿瘤基质中有45例（64.28％）的α-SMA阳性，在94.28％（n = 66）的病例中，在CAF细胞质中鉴定出S100A4。就基质细胞而言，有64例（91.43％）OSCC病例的SHH阳性，GLI-1阳性（31.％）（44.29％）；发现SHH和α-SMA之间呈正相关（p <0.0001，φ= 0.51），以及SHH和S100A4之间（p = 0.087，φ= 0.94）。在初级CAF和NOF中观察到SHH和GLI-1的蛋白表达。尽管发现SHH位于两种细胞类型的细胞质中，但GLI-1仅存在于CAF的细胞核中。我们的结果表明，CAFs不仅是肿瘤基质中HH配体的潜在来源，而且还可能通过核GLI-1激活来响应HH信号。我们进一步观察到，OSCC细胞表达的SHH升高与CAF密度较高有关，从而增强了这些分子发挥的化学引诱作用。φ= 0.94）。在初级CAF和NOF中观察到SHH和GLI-1的蛋白表达。尽管发现SHH位于两种细胞类型的细胞质中，但GLI-1仅存在于CAF的细胞核中。我们的结果表明，CAFs不仅是肿瘤基质中HH配体的潜在来源，而且还可能通过核GLI-1激活来响应HH信号。我们进一步观察到，OSCC细胞表达的SHH升高与CAF密度较高有关，从而增强了这些分子发挥的化学引诱作用。φ= 0.94）。在初级CAF和NOF中观察到SHH和GLI-1的蛋白表达。尽管发现SHH位于两种细胞类型的细胞质中，但GLI-1仅存在于CAF的细胞核中。我们的结果表明，CAFs不仅是肿瘤基质中HH配体的潜在来源，而且还可能通过核GLI-1激活来响应HH信号。我们进一步观察到，OSCC细胞表达的SHH升高与CAF密度较高有关，从而增强了这些分子发挥的化学引诱作用。但也可能通过核GLI-1激活来响应HH信号。我们进一步观察到，OSCC细胞表达的SHH升高与CAF密度较高有关，从而增强了这些分子发挥的化学引诱作用。但也可能通过核GLI-1激活来响应HH信号。我们进一步观察到，OSCC细胞表达的SHH升高与CAF密度较高有关，从而增强了这些分子发挥的化学引诱作用。