5-氮杂脱氧胞苷对乳腺癌细胞MDA-MB-231雌激素受体α表达和增殖侵袭的影响

Effect of 5-Aza-CdR on ERα expression,proliferation and invasion of breast cancer cell line MDA-MB-231

目的:检测5-氮杂脱氧胞苷(5-aza-2’-deoxycytidine,5-Aza-CdR)对乳腺癌细胞MDA-MB-231雌激素受体α(estrogen receptor alpha,ERα)的表达和增殖侵袭的影响。方法:利用细胞生长曲线、MTT法及FCM法检测使用去甲基化药物5-Aza-CdR处理MDA-MB-231细胞后,对MDA-MB-231细胞增殖和周期的影响。通过RT-PCR和免疫细胞化学检测5-Aza-CdR对ERα阴性的乳腺癌细胞MDA-MB-231ERα及基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)表达的影响。通过Boyden小室侵袭实验检测该细胞的浸润能力。结果:5-Aza-CdR能使MDA-MB-231细胞的生长速度明显减慢,细胞周期被阻滞在G0/G1期。5-Aza-CdR能部分恢复ERα阴性乳腺癌细胞MDA-MB-231的ERα表达,并能使MDA-MB-231细胞中MMP-2基因表达下降,细胞的侵袭能力降低。结论:5-Aza-CdR可有效抑制乳腺癌细胞生长并导致细胞周期的阻滞,明显降低乳腺癌细胞的侵袭能力。利用5-Aza-CdR可部分恢复ERα的表达及降低MMP-2的表达,这可能是其引起乳腺癌细胞MDA-MB-231生物学行为改变的机制之一。

Objective：To explore the effect of 5-Aza-2＇ -deoxycytidine （5-Aza-CdR） on ERα expression, proliferation and invasion of human breast cancer cell line MDA-MB-231. Methods：The breast cancer cell line MDA-MB-231 was treated with 5-Aza-CdR. The effect of 5-Aza-CdR on proliferation was evaluated by MTT assay and cell growth curve was drawn. The cell cycle was analyzed by flow cytometry. Immunohistochemistry and RT-PCR were used to determine the changes of ERα expression and matrix metalloproteinase- 2 （MMP-2） expression in the MDA-MB-231 cell after 5-Aza-CdR treatment. The invasion ability of MDA-MB-231 cells was tested by Boyden chamber assay. Results：Compared with the control, the cell growth speed was significantly slowed down and the cell cycle was arrested at G0/G1 phase after 5-Aza-CdR treatment. The expression of ERα gene was partly restored in ERα-negative human breast cancer MDA-MB-231 cell after 5-Aza-CdR treatment. The expression of MMP-2 in MDA-MB-231 cells was down-regulated and the invasion ability of MDA-MB-231 cell was decreased after 5-Aza-CdR treatment. Conclusion ：5-Aza-CdR treatment effectively inhibited cell proliferation, induced cell cycle arrest, and decreased cell invasion ability of breast cancer MDA-MB-231 cells. The findings suggested that the partial restoration of ERα expression and the decrease in MMP-2 expression may be one of the mechanisms for the biological behaviour changes of MDA-MB-231 cells induced by 5-Aza-CdR.