腺样囊性癌两个细胞系体外侵袭能力比较

COMPARATIVE STUDY OF INVASION POTENTIALS IN VITRO WITH TWO HUMAN SALIVARY ADENOID CYSTIC CARCINOMA CELL LINES

目的 研究转移潜能不同的两个腺样囊性癌细胞系体外侵袭能力的差异。方法 选用肺低转移细胞系 Acc- 2和从中筛选出的肺高转移细胞系 Acc- M,利用 MTT法检测与几种基底膜成分的粘附能力 ;划痕法检测运动能力 ;PAGE底物酶谱法检测分泌 型胶原酶的能力 ;改良勃顿小室法观察对重组基底膜的侵袭能力。结果 11小时后Acc- 2细胞系对 3种细胞外基质的粘附率分别是纤维粘连蛋白 (FN) 6 1.8%、层粘连蛋白 (L N) 4 2 .9%和基底膜胶(Matrigel) 6 0 .4% ,Acc- M细胞系的粘附率分别是 FN79.4%、L N71.1%、Matrigel6 9.8%。 2 12小时终止实验时 ,测定 Acc- 2细胞系和 Acc- M细胞系的运动速度分别是1.15μm/h和 2 .2 6μm/h。 3Acc- M细胞系的培养上清中可以检测到 型胶原酶活性 ,而 Acc- 2细胞系的培养上清中检测不到 型胶原酶活性。 48小时终止实验时 Acc- 2细胞系和 Acc- M细胞系对重组基底膜的侵袭率分别是 6 8.0± 4.35和 10 3.2± 5 .6 3。结论 两个腺样囊性癌细胞系的异质型粘附能力、运动能力、分泌 型胶原酶的能力及侵袭重组基底膜能力明显不同 。

Objective To investigate the invasion ability in vitro of two human salivary adenoid cystic carcinoma cell lines with different metastatic potentials.Methods The Acc-2 cell line with low lung metastatic potential and the Acc-M cell line (selected from the Acc-2 cell line) with high lung metastatic potential were used. The adhesive ability to various basement membrance components of the two cell lines was measured by MTT assay, the cell locomotive ability by wound assay, the type Ⅳ collagenase was assessed by PAGE substrate zymography, and the invasion ability by modified Boyden chamber.Results ①The adhesive rates of Acc-2 cell line after 1 h were FN 61.8%、LN 42.9%、MG 60 4 %,and that of Acc-M cell line were FN 79.4%、LN 71.1%、MG 69.8% respectively.②In the wound assay, after 12 hr the locomotive rates of Acc-2 and Acc-M were 1.15 μm/h and 2.26 μm/h.③The type Ⅳ collagenase activity was detected in serum-free supernatant of Acc-M cell line, whereas the Acc-2 cell line was lack of type Ⅳ collagenase activity.④Eight hours after the invasion of the basement membrance matrix, the number of invading cells in high power field of Acc-2 and Acc-M were 68.0±4.35 and 103.2±5.63. Conclusion The results showed that the invasion of Acc-2 and Acc-M was progressive in nature and the heterogeneous adhesive ability,motility,type Ⅳ collagenase activity and invasive capability of these two variants were significantly different and correlative with their metastatic potential.