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目的探讨卡马西平对乳腺癌细胞SKBR-3中人表皮生长因子受体-2(HER-2)表达的影响及其机制。方法分别用卡马西平、赫赛汀、17烯丙胺-17脱甲氧格尔德霉素(17-AAG)处理乳腺癌细胞SKBR-3,通过Western blot检测HER-2蛋白的表达;通过RT-PCR检测HER-2 mRNA的表达;通过免疫沉淀检测热休克蛋白90(HSP90)的分子伴侣功能及乙酰化水平;通过四甲基偶氮唑蓝(MTT)法检测细胞增殖活性。结果10μmol／L卡马西平处理即可降低乳腺癌细胞SKBR-3中HER-2蛋白水平,100μmoL／L卡马西平则在蛋白和mRNA水平均抑制HER-2的表达。卡马西平处理可以使HSP90乙酰化水平升高,并且破坏HSP90的分子伴侣功能。卡马西平对赫赛汀的促HER-2降解作用有协同作用。100μmol/L卡马西平+290μg/ml赫赛汀处理组、100μmol／L卡马西平+1μmol／L 17-AAG处理组的细胞增殖率分别为27．8％±3．0％和26．9％±2．8％,分别明显低于单独赫赛汀或17-AAG处理组(P均<0．01)。结论卡马西平可能通过破坏HSP90的分子伴侣功能、提高HSP90的乙酰化水平,降低了乳腺癌细胞SKBR-3中HER-2的表达水平,并且与赫赛汀和17-AAG有协同作用。卡马西平在肿瘤治疗中有潜在的应用价值。
Objective To investigate the effect of carbamazepine on human epidermal growth factor receptor-2 (HER-2) expression in breast cancer cell line SKBR-3 and its mechanism. Methods The breast cancer cells SKBR-3 were treated with carbamazepine, Herceptin, 17 allylamine-17-demethoxygeldanamycin (17-AAG), and the expression of HER-2 protein was detected by Western blot; The expression of HER-2 mRNA was detected by PCR; the chaperone function and acetylation level of heat shock protein 90 (HSP90) was detected by immunoprecipitation; the cell proliferation activity was detected by MTT assay. RESULTS: Treatment with 10μmol/L carbamazepine reduced HER-2 protein levels in breast cancer cells SKBR-3, and 100μmol/L carbamazepine inhibited HER-2 expression in both protein and mRNA levels. Carbamazepine treatment can increase the level of acetylation of HSP90 and destroy the molecular chaperone function of HSP90. Carbamazepine has a synergistic effect on the promotion of HER-2 degradation by Herceptin. The cell proliferation rates of 100μmol/L carbamazepine+290μg/ml Herceptin treatment group and 100μmol/L carbamazepine+1μmol/L 17-AAG treatment group were 27.8%±3.0% and 26.9 respectively. % ± 2.8%, significantly lower than the Herceptin alone or 17-AAG treatment group (P all <0.01). Conclusion Carbamazepine may reduce the expression of HER-2 in breast cancer cell SKBR-3 by disrupting the molecular chaperone function of HSP90 and increasing the acetylation level of HSP90, and has a synergistic effect with Herceptin and 17-AAG. Carbamazepine has potential application value in the treatment of cancer.