4-乙酰胺苯基维甲酸酯抑制肿瘤细胞侵袭重组基底膜及其作用机理

4 acetamidophenyl retinoate (4 APR) inhibits reconstituted basement membrane invasion by tumor cells and its mechanism

目的研究新维甲类化合物４－乙酰胺苯基维甲酸酯（４－ＡＰＲ）对Ｂ１６－Ｆ１０小鼠黑色素瘤细胞侵袭能力的抑制作用，探讨其作用机理。方法癌细胞侵袭能力用重组基底膜侵袭试验衡量；ＰＡＧＥ底物酶谱方法检测Ⅳ型胶原酶活性；斑点杂交检测ＣＮＥ－２Ｚ细胞ＴＩＭＰ－１ｍＲＮＡ表达；以细胞生长曲线评价药物对Ｂ１６－Ｆ１０细胞的生长抑制作用。结果在１０－５ｍｏｌ／Ｌ和１０－６ｍｏｌ／Ｌ时，４－ＡＰＲ对Ｂ１６－Ｆ１０小鼠黑色素瘤细胞侵袭重组基底膜的抑制率分别为５４．２％和４１．９％，对ＣＮＥ－２Ｚ细胞培养上清中的Ⅳ型胶原酶活性有降低作用。此外，４－ＡＰＲ可抑制Ｂ１６－Ｆ１０细胞与ＬＮ、ＦＮ和Ｍａｔｒｉｇｅｌ的粘附，诱导ＣＮＥ－２Ｚ细胞ＴＩＭＰ－１ｍＲＮＡ表达。结论４－ＡＰＲ抑制Ｂ１６－Ｆ１０细胞侵袭重组基底膜。４－ＡＰＲ的抗侵袭活性与抑制肿瘤细胞的粘附，降低肿瘤细胞培养上清中Ⅳ型胶原酶的活性和（或）诱导肿瘤细胞ＴＩＭＰ－１ｍＲＮＡ表达等有关。

Objective To study the inhibitory effect of new retinoid 4 acetamidophenyl retinoate (4 APR) on the reconstituted basement membrane invasion by B16 F10 mouse melanoma cells and its mechanism. Methods Reconstituted basement membrane invasion assay was used to evaluate invasive ability of cancer cells. Type Ⅳ collagenase was assessed by PAGE substrate zymography. TIMP 1 mRNA expression of human nasopharyngeal carcinoma CNE 2Z cell line was measured by dot blot analysis. Cell growth curve assay was used to examine the growth inhibitory effect of 4 APR on B16 F10 cells. Results 4 APR, at the concentrations of 10 -5 mol/L and 10 -6 mol/L, suppressed the reconstituted basement membrane invasion of B16 F10 mouse melanoma cells by 54.2%, 41.9% and reduced type Ⅳ collagenase activities in the serum free supernatant of CNE 2Z cells. In addition, 4 APR inhibited B16 F10 cell adherence to laminin, fibronectin and Matrigel, and induced CNE 2Z cell TIMP 1 mRNA expression. Conclusion Reconstituted basement membrane invasion of B16 F10 mouse melanoma cells was inhibited by 4 APR. The anti invasion action of 4 APR might be associated with the suppression of tumor cell adhesion ability, the reduction of type Ⅳ collagenase activity in tumor cell culture supernatant and/or the induction of tumor cell TIMP 1 mRNA expression.