硫酸乙酰肝素对小鼠乳腺肿瘤细胞Caspase-3、bcl-2的表达及Caspase-3活性影响

Effects of heparan sulfate on Caspase-3 activity and expressions of Caspase-3,bcl-2 in mice breast tumor cells

目的:探讨硫酸乙酰肝素(heparan sulfate,HS)对乳腺肿瘤细胞Caspase-3、bcl-2表达及Caspase-3活性的影响。方法:(1)培养人乳腺上皮癌MCF-7细胞株,生长达汇合期及汇合后期时收集培养液,提取其中的硫酸乙酰肝素链。(2)建立C3H小鼠移植乳腺癌模型,随机分3组,即生理盐水(NS)组、HS组、阿霉素(adriamycin,ADR)组。第2天腹腔注射给药,给药结束第2天处死小鼠,完全剥离瘤体,称瘤重,计算抑瘤率。(3)免疫组化法测定Caspase-3、bcl-2的表达,Caspase-3活性检测试剂盒检测Caspase-3的活性。结果:(1)HS对小鼠肿瘤生长有抑制作用,HS和ADR组的肿瘤生长抑制率分别为29.98%和46.87%;(2)HS能减少肿瘤bcl-2表达,各组的阳性细胞数为,HS组34.6±2.55、ADR组46.0±3.24、NS组71.5±2.87;(3)HS能促进小鼠肿瘤细胞Caspase-3表达并能增强其生物活性,阳性细胞数为,HS组56.12±2.16、ADR组35.50±2.39、NS组19.75±1.98;各组的Caspase-3活性分别为HS组19.89±1.36、ADR组18.16±0.25、NS组13.15±0.90。结论:HS能通过减少C3H小鼠乳腺癌细胞bcl-2的表达,增加Caspase-3表达和活性来促进肿瘤细胞凋亡,发挥其抗肿瘤作用。

Objective To detect the effects of heparan sulfate（HS）on the Caspase-3 and bcl-2 expressions and Caspase-3 activity in mice breast tumor cells. Methods （1）HS was extracted from the conditional medium in which human breast cancer cells （MCF-7）grew to confluence or past confluence. （2）Transplanted breast cancer models of C3H mice were established and divided into 3 groups[normal sodium（NS） group, HS group, adriamycin（ADR） group] randomly. From the next day, the explanted animals were administered different agents intraperitoneally. At the next day of administration, the mice were killed, tumors were weighed to calculate tumor inhibition ratio. （3）The expressions of Caspase-3 and bcl-2 were detected by immunohistochemistry as well as the Caspase-3 activity by Caspase-3 activity kit. Results HS could inhibit the growth of C3H mouse breast cancer, the inhibitory rate of HS was 29.98%, by contrast, the inhibitory rate of adriamycin was 46.87%. （2）HS could reduce the expression of bcl-2, HS group 34.6 ± 2.55, ADR group 46.0 ± 3.24, NS group 71.5 ± 2.87. HS could enhance Caspase-3 expression, Caspase-3 expressions in each group respectively were HS 56.12 ± 2.16, ADR 35.50 ± 2.39, NS 19.75 ± 1,98. Furthermore it could increase Caspase-3 activity, 19.89 ±1.36 for HS group, 18.16 ± 0.25 for ADR group and 13.15 ± 0.905 for NS group respectively. Conclusion HS can induce tumor cells apoptosis by increasing the expression of Caspase-3, enhancing Caspase-3 activity and decreasing the expression of bcl-2.