赖氨匹林对B16黑色素瘤的增殖抑制及促凋亡作用

目的探讨赖氨匹林(aspisol)在体外和体内对小鼠B16黑色素瘤细胞的增殖抑制和诱导凋亡作用。方法利用MTT法和流式细胞术检测赖氨匹林对B16细胞的增殖抑制和诱导凋亡作用。用细胞悬液法将B16黑色素瘤细胞接种于小鼠前肢腋窝皮下,制备可移植肿瘤模型。次日给予不同浓度的赖氨匹林腹腔注射,每天1次,共28天,用达卡巴嗪(dacarbazine,DTIC)和生理盐水分别作阳性对照和阴性对照。计算aspisol的抑瘤率;原位凋亡检测法(TUNEL)检测赖氨匹林对肿瘤细胞凋亡的影响;免疫组化法检测aspisol对小鼠肿瘤组织的Survivin、C-erbB-2表达的影响。结果Aspisol可抑制B16细胞增殖,最大抑制率为(68.78±1.27)%,诱导B16细胞凋亡,最大凋亡率为15.8%。200、400、800mg·kg-1aspisol对小鼠黑色素瘤的抑瘤率分别为15.0%、32.3%、49.4%,40mg·kg-1DTIC的抑瘤率为51.4%。各给药组肿瘤细胞均呈现明显凋亡形态改变,不同浓度aspisol均明显下调小鼠肿瘤组织的Survivin、C-erbB-2表达。结论Aspisol在体外和体内能够抑制小鼠B16黑色素瘤增殖和诱导凋亡,其机制可能与抑制Survivin、C-erbB-2表达有关。

Inhibitory and apoptosis-inducing effects of aspisol on proliferation of B16 melanoma

Aim To study the inhibitory and apoptosis-inducing effects of aspisol on proliferation of B16 melanoma in vitro and in vivo. Methods The effect of aspisol on the proliferation of B16 cells was analyzed by MTT assay; its effect on cell apoptosis was measured by flow cytometry. The suspension of melanoma cells was injected subcutaneously into forelimb axillas of C57BL/6J mice to establish xenograft models. From the next day, the mice received intraperitoneal injection of aspisol in different concentrations once a day for 28 days; the mice received injection of dacarbazine (DTIC) were used as positive controls,and received injection of normal saline (NS) were used as negative controls. The inhibition rate of tumor growth was calculated .The apoptosis rate was detected by TUNEL assay. The expression of Survivin and C-erbB-2 was detected by immunohistochemistry. Results Aspisol inhibited the proliferation of B16 cells, with the inhibition rate up to (68.78±1.27)%, and induced the apoptosis with the highest apoptosis rate up to 15.8%.The inhibition rate of tumor growth was 15.0% in 200 mg·kg-1 aspisol group, 32.3% in 400 mg·kg-1 aspisol group, 49.4% in 800 mg·kg-1 aspisol group,and 51.4% in 40 mg·kg-1 DTIC group. Typical apoptotic morphologic changes were seen in the 4 groups. Survivin and C-erbB-2 expression was significantly lower in aspisol groups than in NS group. Conclusion Aspisol could inhibit proliferation and induce apoptosis of B16 melanoma cells in vitro and in vivo, which may be correlated to down-regulation of Survivin and C-erbB-2.