表皮生长因子受体酪氨酸激酶抑制剂厄洛替尼联合山奈酚诱导卵巢癌细胞凋亡的协同机制

Synergetic effect of epidermal growth factor receptor tyrosine kinase inhibitor kaempferol in combination with tarceva on apoptosis of ovarian cancer cells

目的:探讨厄洛替尼和山奈酚对表皮生长因子受体酪氨酸激酶(EGFR-TPK)的抑制作用,并阐明二者对卵巢癌细胞生长的协同抑制作用机制。方法:培养人卵巢癌SKOV-3细胞,将细胞分为空白组、山奈酚组和厄洛替尼组,将不同浓度山奈酚与厄洛替尼加入到EGFR-TPK和人卵巢癌SKOV-3细胞中,ELISA法检测EGFR-TPK的活性,采用Transwell小室法检测各组SKOV-3细胞侵袭转移能力,采用酶标仪检测各组SKOV-3细胞凋亡蛋白caspase-3活性,MTT法检测各组SKOV-3细胞的生长抑制率及山奈酚联合厄洛替尼对SKOV-3细胞的生长抑制作用。结果:山奈酚和厄洛替尼对EGFR-TPK的半数抑制浓度(IC50)分别为(2.33±0.32)g·L-1和(1.75±0.18)g·L-1,二者比较差异无统计学意义(P>0.05)。与空白组比较,山奈酚和厄洛替尼组穿膜细胞数减少(P<0.05或P<0.01),SKOV-3细胞中凋亡蛋白caspase-3活性明显增加(P<0.05或P<0.01)。随着山奈酚和厄洛替尼浓度增加,SKOV-3细胞生长抑制率不断增加,与空白组比较差异均有统计学意义(P<0.05或P<0.01)。结论:山奈酚联合厄洛替尼对人卵巢癌SKOV-3细胞具有协同抑制作用,并对SKOV-3细胞转移起到抑制作用。

Objective： To study the inhibitory effect of epidermal growth factor receptor tyrosine kinase （EGFR- TPK）, and to clarify the mechanism of their synergetic inhibitory effects on the growth of ovarian cancer cells. Methods： The human ovarian cancer SKOV 3 cells were cultured and divided into blank group, kaempferol group and tarceva group. The activities of EGFR-TPK were detected by ELISA method. The invasion abilities of SKOV-3 cell lines were determined by Transwell assay. The caspase-3 activities of SKOV-3 cells were measured. The inhibitory rates of growth of SKOV-3 cells were determined using MTT method. Results The half inhibitory concentrations （IC50） of kaempferol and tarceva were （52.33±0.32） g · L-land （1.75±0.18） g · L-1 , there was no significant differences between them （P〈0.05）. The inhibitory rates of growth of SKOV-3 cells were increased with the increasing of doses of kaempferol and tarceva, and there were significant differences compared with blank group （P〈0. 05 or P〈0.01）. The caspase-3 activities of SKOV-3 cells of in kaempferol and tarceva groups were significantly increased compared with blank group （P〈0.05）. Conclusion.. Kaempferol in combination with tarceva has a synergetic effect on the growth of ovarian cancer SKOV-3 cells.