AGPS在侵袭性纤维瘤侵袭机制中的研究

Role of alkyldihydroxyacetonephosphate synthase in invasive mechanism of aggressive fibromatosis

目的探索烷基甘油磷酸合成酶（alkyldihydroxyaeetonephosphate synthase，AGPS）在侵袭性纤维瘤（aggressive fibromatosis，AF）侵袭能力中发挥的作用；观察异硫氰酸苄酯（Benzyl isothiocyanate，BITC）对AF细胞AGPS的表达及侵袭的影响。方法收集30例我院病理确诊为AF的标本组织，其中男18例，女12例，患儿平均年龄6岁，原发13例，复发17例，臀部21例，大腿7例，前臂2例，瘤体平均大小336cm3，收集6例来源于内固定取出术的瘢痕组织，提取培养两例原代肿瘤细胞（AF1及AF2），在组织学水平以瘢痕组织为对照通过免疫组化方法，细胞水平以人正常成纤维细胞（HFF）为对照，通过Westernblot方法分别观察AGPS在组织及细胞水平的表达；运用SiR—NA干扰技术抑制AF细胞AGPS的表达后采用Wound healing及Transwell观察AF细胞迁移及侵袭能力的改变；5μm／LBITC干预AF细胞，等量二甲基亚砜（DMSO）作为对照，Westernblot测定AF细胞AGPS的表达、Woundhealing及Transwell观察AF细胞迁移及侵袭能力的改变。结果AF组织及细胞中AGPS的表达明显高于对照组（P组织=0．001、PAF1=0．006和PAF2=0．003）；SiRNA沉默AGPS表达后AF细胞的迁移（PWound healing〈0．01，PMigration=0．000）以及侵袭（P=0．000）能力明显减弱；BITC可以降低AGPS的表达（P=0．029），并抑制AF细胞的迁移（Wound healing P值分别为0．039（12h）及0．004（23h），migration P=0．000）以及侵袭能力（P=0．000）。结论AGPS在AF的侵袭力中发挥一定的作用，为治疗AF提供新的药物靶点。

Objective To explore the role of alkyldihydroxyacetonephosphate synthase （AGPS） in the invasiveness of aggressive fibromatosis （AF） and observe the effect of benzyl isothiocyanate （BITC） upon the expression and invasion of AGPS in AF cells. Methods A total of 30 cases of AF samples were collected and diagnosed by pathologic examination. There were 18 boys and 12 girls with an average age of 6 years. There were primary （n= 13） and recurrent （n-＇- 17） cases. The sites were located on hip （n = 21）, thigh （n = 7） and forearm （n = 2）. The average size was 336 cm^3. Six cases of scar samples were harvested intraoperatively during internal fixation for cultivating two cases of primary tumor cells （AF1 ＆ AFE）. At the histological level, as compared with scar by immunohistochemistry, cell level human normal fibroblasts （HFF） was used as control to observe the expression of AGPS at histological and cell levels respectively. Wound healing and Transwell assays were used for observing the changes of migration and invasiveness after AGPS silencing by SiRNA. The expression of AGPS was detected by Western blot. Wound healing and Transwell assays were used for observing the changes of migration and invasiveness after a pre - treatment of 5 m/LBITC. And the same volume of dimethyl sulfoxide （DMSO） was used as control. Results The expressions of AGPS were significantly higher in AF tissue and cells those that in control group [P = 0. 001 （tissue）, 0. 006 （AF1） ＆ 0. 003 （AF2）]. The migration （wound healing P〈0. 01, migration P = 0. 000） and invasiveness （P = 0. 000） of AF cells decreased significantly after AGPS silencing by SiRNA. BITC could reduce the expression of AGPS （P = 0. 029） and inhibit the migration （wound healing P = 0. 039 （12 h） and 0. 004 （23 h） respectively, migration P = 0. 000） and invasion （P = 0. 000） of AF cells. Conclusions AGPS plays some role in the invasiveness of AF and provides a new drug therapeutic target for AF.