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[目的]观察胰岛素样生长因子(IGF)受体通路对卵巢癌休眠肿瘤细胞启动增殖的调控机制。[方法]采用免疫细胞化学法检测SKOV3-PKH26hi细胞IGF-1、IGF-2和IGF-1R的表达,酶联免疫吸附测定法(ELISA)测定细胞培养液中IGF-1和IGF-2的浓度,细胞计数和MTT法测定加入外源性IGF-1对SKOV3-PKH26hi细胞增殖的影响,流式细胞术检测细胞IGF-1R和Ki-67的表达、细胞周期和凋亡率,Western blot方法检测细胞p-AKT、p-GSK3β及cyclin B1的表达。[结果]SKOV3-PKH26hi细胞均可以表达IGF-1、IGF-2和IGF-1R;培养液中IGF-1和IGF-2的浓度随培养时间的延长显著升高(P<0.05);IGF-1对SKOV3-PKH26hi细胞能发挥促增殖作用;SKOV3-PKH26hi细胞IGF-1R、Ki-67、p-AKT、p-GSK3β、cyclin B1的表达量随着培养时间的延长而显著增高(P<0.05);随着培养时间的增加SKOV3-PKH26hi细胞G0/G1和G2/M期细胞显著减少(P<0.05),S期细胞显著增加(P<0.05),不同时间点的凋亡率差异无统计学意义(P>0.05)。[结论]SKOV3-PKH26hi细胞存在IGF自分泌,IGF受体通路能够调控卵巢癌休眠细胞的增殖。
[Objective] To investigate the regulatory mechanism of insulin-like growth factor (IGF) receptor pathway on the initiation and proliferation of dormant tumor cells in ovarian cancer. [Methods] The expressions of IGF-1, IGF-2 and IGF-1R in SKOV3-PKH26hi cells were detected by immunocytochemistry. The concentrations of IGF-1 and IGF-2 in the cell culture medium were measured by enzyme linked immunosorbent assay (ELISA) The cell proliferation and proliferation of SKOV3-PKH26hi cells treated with exogenous IGF-1 were measured by cell counting and MTT assay. The expression of IGF-1R and Ki-67, cell cycle and apoptosis rate were detected by flow cytometry and Western blot Cell p-AKT, p-GSK3β and cyclin B1 expression. [Results] The expression of IGF-1, IGF-2 and IGF-1R in SKOV3-PKH26hi cells was significantly increased (P <0.05). The concentration of IGF- 1 could increase the proliferation of SKOV3-PKH26hi cells. The expression of IGF-1R, Ki-67, p-AKT, p-GSK3β and cyclin B1 in SKOV3-PKH26hi cells was significantly increased with the prolongation of culture time ), The number of cells in S phase (P <0.05) and the number of apoptotic cells in SKOV3-PKH26hi cells at different time points were not statistically different (P <0.05) Significance (P> 0.05). [Conclusion] There is IGF autocrine in SKOV3-PKH26hi cells, IGF receptor pathway can regulate the proliferation of dormant cells in ovarian cancer.