【摘要】 目的　探讨αvβ6整合素介导的细胞与细胞外基质的黏附 ,及其对顺铂诱导的卵巢癌细胞凋亡的影响。方法　采用流式细胞仪测定卵巢癌细胞株SKOV3、AO、3AO细胞表面αvβ6整合素的表达 ,并利用酶联免疫吸附实验、吖啶橙 溴化乙锭双荧光染色法分析顺铂诱导的细胞凋亡及αvβ6整合素对细胞凋亡的影响。结果　SKOV3、AO、3AO细胞均表达αvβ6整合素 ,其相对荧光指数 (FI)均为 1 0 3± 0 0 1。顺铂可以诱导SKOV3、AO、3AO细胞凋亡 ,经顺铂 (10 μg/ml)作用 4 8h后 ,酶联免疫吸附实验结果显示 ,生长于αvβ6整合素配体———纤维粘连蛋白 (FN)上的细胞的富集系数 (EF)分别为 2 11± 0 0 4、2 15± 0 0 6、2 11± 0 0 4 ,明显低于生长于非整合素配体———多聚赖氨酸 (polylisin)上的细胞 (分别为 3 5 1± 0 0 3、3 5 5± 0 0 4、3 6 6± 0 0 4 ,P <0 0 1) ;吖啶橙 溴化乙锭双荧光染色法结果显示 ,生长于FN上的细胞凋亡率分别为 (5 0± 0 7) %、(5 0± 0 7) %、(6 0± 0 7) % ,明显低于生长于polylisin上的细胞凋亡率[分别为 (2 8 0± 0 7) %、(2 8 0± 0 7) %、(2 9 0± 0 7) % ,P <0 0 0 1],用αvβ6整合素的特异性阻断抗体封闭过的细胞再次生长于FN上时 ,细胞凋亡率 [分别为 (15 0± 0更多还原

【Abstract】 Objective To investigate the effects of αvβ6 integrin-mediated cell adhesion on cisplatin(DDP) induced apoptosis in ovarian cancer cell lines. Methods The expression of integrin αvβ6 in ovarian cancer cell lines SKOV3, AO, and 3AO was analyzed by flow cytometry, and the apoptosis induced by DDP was measured by enzyme-linked immunosorbent assay(ELISA) method and acridine orange-ethidium bromide (AO-EB) double fluorescent dye staining. Results Ovarian cancer cell lines SKOV3, AO and 3AO all expressed integrin αvβ6. Treated by different concentrations of DDP, SKOV3, AO and 3AO displayed significant apoptosis status compared with the control. However,after cultured in medium containing 10 μg/ml DDP for 48 hours, SKOV3, AO and 3AO planted on fibronectin (FN)-ligand of αvβ6 integrin, had lower enrichment factor (EF), compared with those planted on non-integrin ligand polylisin measured by ELISA method(2.11±0.04 vs 3.51±0.03,2.15±0.06 vs 3.55±0.04,2.11±0.04 vs 3.66± 0.04, P <0.01). Similarly, by AO-EB double fluorescent dye staining, lower percentages of apoptosis were found in cells planted on FN (5.0±0.7)%,(5.0±0.7)%,(6.0±0.7)%,than on polylisin(28.0±0.7)%,(28.0±0.7)%,(29.0±0.7)% ( P <0.001). When cells preincubated with integrin αvβ6 blocking antibody were planted on FN, the percentages of apoptosis rose again,(15.0±0.7)%,(16.0±0.7)%,(15.0±0.7)% ( P <0.01). Conclusions The de novo expression of αvβ6 integrin occurs in ovarian cancer cell lines SKOV3,AO, and 3AO. αvβ6 integrin-mediated cell adhesion to FN can inhibit the effect of cisplatin-induced apoptosis. These results suggest that inhibiting apoptotic cell adhesion may improve drug resistance in ovarian cancer cell lines.更多还原