黄连素对乳腺癌细胞生长、迁移和放射敏感性的影响

Impacts of berberine on the growth, migration and radiosensitivity of breast cancer cells

目的研究黄连素对乳腺癌细胞生长、迁移和放射敏感性的影响。方法选取人乳腺癌细胞MDA—MB-231和MCF-7，用MTT法检测细胞的生长和增殖；划痕愈合法观察细胞迁移能力；细胞流式技术测定细胞周期改变；磷脂酰丝氨酸外翻分析法检测细胞凋亡；Westernblot法测定蛋白表达水平；细胞克隆形成法观察黄连素与辐射的联合作用。结果黄连素明显抑制人乳腺癌MCF-7和MDA-MB-231细胞的生长，并显现剂量．效应和时间-效应关系，MCF-7和MDA-MB-231细胞对黄连素表现出同样的敏感性。黄连素可以诱导剂量依赖性的G0／G1期细胞阻滞，40μmol／L的黄连素处理后MDA-MB-231和MCF-7细胞早期凋亡分别高达86．63％和66．62％，与未处理的对照组相比，差异均有统计学意义（t=75．15和43．75，P〈0．01）。黄连素明显降低了细胞周期调节相关蛋白CyclinB1和抑凋亡蛋白Bcl-2的表达水平，而增加了促凋亡蛋白Bax和Caspase-3的表达水平，但不影响细胞周期调节蛋白CyclinD1的表达水平。低剂量（≤5μmol／L）黄连素可以明显降低细胞的迁移能力。采用单靶多击模型拟合曲线发现，与单独照射组相比，5μmol／L黄连素预处理4h后MDA-MB-231细胞和MCF-7细胞的辐射增敏比分别为1．12和1．22。结论黄连素通过调节细胞凋亡和细胞周期阻滞抑制乳腺癌细胞生长和迁移，并可提高乳腺癌细胞的放射敏感性。

Objective To study the impacts of berberine on the growth, migration and radiosensitivity in human breast cancer cells. Methods MTT assay was used to evaluate cell growth. In vitro scratch migration assay was used to determine cell migration. Annexin V assay was used to detect cell apoptosis. The distribution of cell cycle was evaluated by flow eytometry assay. Colony formation assay was used to detect the influence of berberine on cell radiosensitivity. Western blot assay was employed to measure protein expression. Results Berberine inhibited cell growth and migration in two human breast cancer cell lines, MCF-7 and MDA-MB-231, in a dose-and time-dependent manner. Furthermore, berberine resulted in a cell cycle G0/GI arrest. Compared with control, the early apoptosis in MDA-MB- 231 and MCF-7 cells treated with 40 p.mol/L of berberine was as high as 86.6% and 66. 6% （t = 8.79, 10. 32, P 〈 O. 01 ） , respectively. Berberine caused a dose-dependent increase in Bax and Caspase-3 protein expressions, but did not change Cyclin D1 protein expression, while suppressed the expressions of Cyclin B1 and Bcl-2 protein. As analyzed with multi-target click model fitting curves, the SERo0 of berberine-treated cells were 1.12 and 1.22 for MDA-MB-231 and MCF-7cells respectively at the dose Do of X-rays. Conclusions The berberine inhibited the growth and migration of breast cancer cells via apoptosis induction and cell cycle arrest. Moreover, berberine increases cell sensitivity to X-ray irradiation.