紫龙金对人乳腺癌细胞系MCF-7增殖及凋亡的影响

Effects of Zilongjin on Proliferation and Apoptosis of Human Breast Cancer Cell Line MCF-7

目的观察复方中药紫龙金对人乳腺癌细胞系MCF-7增殖的抑制作用及对其凋亡的诱导作用,探讨紫龙金治疗乳腺癌的作用机制。方法依据培养液紫龙金浓度的差异,实验共设4组:(1)对照组:不加紫龙金干预的1640培养液;(2)紫龙金低浓度组:1.5 mg生药/mL紫龙金培养液;(3)紫龙金中浓度组:3 mg生药/mL紫龙金培养液;(4)紫龙金高浓度组:6 mg生药/mL紫龙金培养液。采用细胞计数法绘制生长曲线及四甲基偶氮唑蓝(MTT)比色法检测不同浓度紫龙金对MCF-7细胞增殖能力的影响,并应用流式细胞术、Hoechst 33342核染色法及DNA梯度形成法测定紫龙金诱导凋亡的作用。结果(1)与对照组比较,紫龙金各剂量组的细胞计数在各时间点(24、48、72、96、120、144 h)明显减少,其差异均有统计学意义(P<0.05);紫龙金各剂量组间的生长抑制率差异除低、中浓度组间在24、72 h无统计学意义外(P>0.05),各剂量组间两两比较在不同时间点(24、48、72、96、120、144 h),其差异均有统计学意义(P<0.05)。紫龙金对MCF-7细胞的增殖抑制作用呈时间依赖性和剂量依赖性;(2)紫龙金使细胞阻滞在G_0/G_1期;(3)中、高浓度紫龙金诱导MCF-7细胞凋亡,并形成DNA ladder。结论紫龙金能抑制人乳腺癌细胞MCF-7细胞增殖,并诱导细胞凋亡,从而起到抗肿瘤的作用。

Objective To study the effect of Zilongjin （ZLJ, a composite Chinese drug） on proliferation and apoptosis of human breast carcinoma cell line MCF-7. Methods MCF-7 cells were randomly divided into four groups depending on the culture solution used, the control group, cultured with 1640 medium not contained ZLJ; and the three ZJL groups cultured with medium contained low （1.5 mg/mL）, moderate （3 mg/mL） and high （6 mg/mL） dosage of ZLJ crude drug respectively. The changes of cell proliferation were assessed by cell growth curve assay and methyl thiazolyl tetrazolium （MTT） assay. And the cell apoptosis was analyzed by flow cytometry, Hoechst 33342 staining and DNA ladder assay. Results Compared with that in the normal control, the counts of cells in the three ZLJ groups were decreased significantly （ P 〈 0.05） at such time point as 24, 48, 72, 96, 120, and 144 h. Furthermore, apart from the comparation of the growth inhibition rate between the low and moderate dosage group at 24 and 72 h which were found to be no significant difference （ P 〉 0.05）, the comparations of that among the three ZLJ groups appeared to be significant difference （ P 〈 0.05）. The inhibitory effect of ZLJ on cell proliferation of MCF-7 was time- and dose-dependent; it could retard cells in G0/G1 cell phase ; apoptosis of MCF-7 cell was induced by modrate and high dosage of ZLJ with revealing of apoptotic body and DNA ladder formation. Conclusion ZLJ shows cell proliferation inhibitory and apoptosis inducing effects on human breast cancer cell line MCF-7, and thus to realize its anti-tumor action.