全反式维A酸联合辛二酰苯胺异羟肟酸对乳腺癌MCF-7细胞的作用

Effect of all trans-retinoic acid combined with suberoylanilide hydroxamic acid on breast cancer cells MCF-7

目的诱导和促使肿瘤细胞向正常细胞分化是一种新的肿瘤治疗方式,目前临床上诱导分化药物多应用于白血病的治疗,因此开展实体肿瘤方面的体外研究势在必行。本实验将全反式维A酸(ATRA)和辛二酰苯胺异羟肟酸(SAHA)联合用于乳腺癌MCF-7细胞,探讨两药联合对细胞增殖的影响以及联合用药是否具备协同增效的作用。方法培养人乳腺癌MCF-7细胞,待细胞进入对数生长期,加入一定浓度的ATRA和(或)SAHA,倒置显微镜下观察细胞形态变化、MTT比色法检测增殖抑制作用。吸光度值及抑制率比较采用重复测量资料的方差分析,采用LSD进行两两比较。结果倒置显微镜下加药组形态发生改变,两药联合组较单药组明显,且随时间延长作用更加显著;24 h时间点A1、B1组吸光度值与对照组比较差异无统计学意义(P=0.092),48 h时间点B1组与对照组比较差异无统计学意义(P=1.243)。72 h、96 h、120 h时间点各实验组吸光度均值均小于对照组(分组差异有统计学意义,F=320.648,P=0.000;不同时间点吸光度值差异也有统计学意义,F=219.245,P=0.000;分组与时间具有交互作用,F=117.962,P=0.000)。单药组增殖抑制率较对照组升高,两药联合组高于单药组,差异有统计学意义(分组:F=462.792,P=0.000;不同时间点:F=315.024,P=0.000;交互作用:F=179.682,P=0.000);两药联合作用q值在A1+B1组、A2+B1组24 h、48 h时间点均大于1.15,表现为协同作用,其余q值均大于0.85小于1.15,表现为相加作用。结论 ATRA和SAHA单独或联合作用于乳腺癌MCF-7细胞均可抑制其增殖,两药联合具有协同作用。

Objective Induction and differentiation of tumor cells is a new therapy of cancer. Nowadays, differentiation inducing agents were widely used in treatment of acute lymphoblastic leukemia, therefore it is imperative to develop the investigation in vitro of solid tumors. This study was to combine all trans-retinoic acid(ATRA) and suberoylanilide hydroxamic acid(SAHA) on the proliferation of MCF-7 to investigate whether the combination of the two drugs have synergistic effect. Methods Human breast cancer cell line MCF-7 was cultured in vitro. When the cells entered the logarithmic growth phase, a certain concentration of ATRA(or) SAHA were added into the petri dishes. Morphological changes of cells were observed under inverted microscope, the anti-proliferative effect was measured by MTT assay. Absorbance value and inhibition rate were compared by repeated measurement data, using LSD for multiple comparison. Results Inverted microscope observation showed that, morphological changes of cells in ATRA combined with SAHA group was obvious than ATRA or SAHA group, and with the extension of time the effect was more remarkable. The absorbance value of A1 and B1 group was not statistically significant compared with the control group at 24 h(P=0.092). Compared with the control group, the B1 group had no statistical significance at 48 h(P=1.243). The average absorbance of each experimental group was less than that of the control group at 72 h, 96 h, 120 h(groups: F=320.648, P=0.000; different time-point: F=219.245, P=0.000; interaction: F=117.962, P=0.000). The proliferation inhibition rate of single drug group was higher than that of the control group, and the ATRA combined with SAHA group was significantly higher than that in the single drug group(groups: F=462.792, P=0.000; different time-point: F=315.024, P=0.000; interaction: F=179.682, P=0.000); q values in A1+B1 group and A2+B1 group of combined effects of two drugs at 24 h and 48 h were greater than 1.15, performed as synergetic effect, q values in other groups and time points were greater than 0.85, performed as additive effects. Conclusions ATRA and SAHA can inhibit the proliferation of MCF-7 of breast cancer cells. In inhibiting the proliferation of breast cancer cells, the combination of the two drugs has a synergistic effect.