摘要
[目的]观察消瘤汤含药血清对人肝癌细胞HepG2凋亡相关因子表达的影响,进一步证实该方对肿瘤细胞的抑制作用,并初步探讨其诱导肝癌细胞凋亡的作用机制。[方法]给予昆明种小鼠中药煎剂灌胃3d后,制备消瘤汤鼠含药血清,将其作用于HepG2细胞,用四氮唑盐法(MTT法)检测含药血清对HepG2细胞增殖的抑制作用,通过流式细胞技术测定肿瘤细胞的凋亡情况,应用免疫组化法检测半胱氨酸蛋白酸3(Caspase-3)、B细胞淋巴瘤2(Bcl-2)蛋白表达水平。[结果]消瘤汤鼠含药血清作用于HepG2细胞72h后,对肿瘤细胞生长有明显抑制作用。流式细胞仪检测可见含药血清组在细胞G1/G0期前出现明显的凋亡峰,即"亚二倍体峰"。凋亡率为22.3%,明显高于对照组的1.05%(P<0.05)。免疫组化结果表明Caspase-3在含药血清干预后,表达明显增加,而Bcl-2在含药血清干预后,表达明显降低。[结论]消瘤汤可能通过激活Caspase-3及抑制Bcl-2的活性,从而诱导肝癌细胞凋亡,有效地抑制肝癌细胞生长,达到抗肿瘤的作用。
[-Objective]To investigate the effects of the serum containing anti-cancer prescription （ACP） on expression of apoptosis associated factors in human hepatoma carcinoma cell HepG2 to prove the inhibitory effect of serum containing ACP on the proliferation of HepG2 and to explore its effective mechanism of cell apoptosis, [Methods]The rats were administrated with the herbs and then the blood was collected to prepare serum containing ACP, the serum containing ACP was added to culture HepG2 cells. The MTT assay was used to examine the proliferation inhibitory activity and the flow cytometry was used to assay the apoptosis. Immunohistochemistry was used to detect the expression of Caspase-3 and Bcl-2 in the level of protein.[Results]The serum containg ACP has an obvious inhibitory effect on the proliferation of HepG2 for 72 hours. The results of the flow cytometry revealed that there was a remarkable apoptotic peak before G0-G1 phase after Dretreatment with the serum, hypodiploid peak, the rate of apoptosis was 22. 3%,which was higher than the control（1.05%） obviously. The results of the immunohistochemistry demonstrated the expression of Caspase-3 was increased, compared with the low expression of Bcl- 2,after treatment with the serum containg ACP. [Conclusion]The serum containing ACP has a significant inhibitory effect on the proliferation of HepG2. The mechanism of inducing cell apoptosis may be relevant to stimulate Caspase-3 and suppress Bcl-2.
出处
《中国中西医结合消化杂志》
CAS
2009年第3期171-174,共4页
Chinese Journal of Integrated Traditional and Western Medicine on Digestion
