中文摘要：

目的：探讨中药单体淫羊藿苷（icarrin,ICA）、黄芩苷（baicali,BAI）联合化疗药多柔比星（doxorubicin,又称ADM）抑制肝癌HepG2细胞增殖诱导配体（proliferation-inducing ligand,APRIL）的表达,进而抑制肿瘤细胞的增殖和逆转肿瘤细胞免疫逃逸的作用和机制。方法：MTT法检测25μg/ml ICA、200μg/ml BAI、2μg/ml ADM以及12.5μg/ml ICA＋1μg/ml ADM、100μg/ml BAI＋1μg/ml ADM等5个用药处理组与未用药对照组HepG2细胞的增殖活性;RT-PCR检测APRIL及其受体HSPG、凋亡相关蛋白Bcl-2的表达水平;MTT法检测CD3AK（anti-CD3 antibody induced activated killer cells）对用药处理组和未用药对照组HepG2细胞的杀伤活性。结果：ICA、BAI以及ADM对HepG2细胞增殖具有明显的抑制作用,且呈时间依赖效应;ICA、ADM、ICA＋ADM、BAI＋ADM组作用96 h抑制率最高,分别为（29.30±6.62）%、（63.60±1.35）%、（99.03±0.10）%、（98.89±0.18）%（均P〈0.01）。HepG2细胞高表达APRIL及其受体HSPGmRNA。ICA、BAI以及ADM下调HepG2细胞APRILmRNA以及凋亡相关蛋白bcl-2 mRNA的表达水平。ICA、BAI、ADM以及ICA＋ADM、BAI＋ADM能增加HepG2细胞对CD3AK细胞杀伤的敏感性,杀伤率由对照组的（30.00±4.50）%分别增加到（97.23±5.11）%、（93.12±9.88）%、（60.45±5.71）%、（43.87±8.2）%、（47.25±2.68）%（均P〈0.01）;且经CD3AK作用后,HepG2细胞bcl-2 mRNA的表达进一步下调。结论：ICA、BAI联合ADM抑制HepG2细胞APRIL、bcl-2 mRNA的表达,进而抑制肿瘤细胞增殖;同时增强其对CD3AK细胞的杀伤敏感性,逆转肿瘤细胞的免疫逃逸。

英文摘要：

Objective： To study the inhibitory effect of icarrin （ICA） combined with baicali （BAI） and doxorubicin （ADM） on expression of APRIL in HepG2 cells, so as to study their inhibitory effect on HepG2 cell proliferation and their reversal effect on immune escape and the related mechanism. Methods： MTT assay was used to explore the effects of 25 μg/ml ICA, 200 μg/ml BAI, 2 μg/ml ADM, 12.5 μg/ml ICA ＋ 1 μg/ml ADM and 100 μg/ml BAI ＋ 1 μg/ml ADM on proliferation of HepG2 cells; untreated cells were taken as control. RT-PCR was used to examine the expression of APRIL, its receptor HSPG and apoptosis-associated gene bcl-2. MTT assay was also used to examine cytoxicity activity of CD3AK（ anti-CD3 antibody induced activated killer cells） effector cells on HepG2 cells treated and untreated with drugs. Results ： ICA, BAI and ADM all had obvious inhibitory effect on the proliferation of HepG2 cells, and this inhibition was time dependent. The most severe inhibitions were seen 96 hours after treatment with ICA, ADM, ICA ＋ ADM and BAI ＋ ADM, with the inhibitory rate being （29.30 ±6.62）%, （63.60 ± 1.35）%, （99.03 ±0. 10）% and （98.89 ± 0. 18）%, respectively（ all P 〈0.01 ）. APRIL and its receptor HSPG were strongly expressed in HepG2 cells. ICA, BAI and ADM downregulated APRIL and bcl-2 mRNA expression compared with untreated HepG2 cells. ICA, BAI, ADM, ICA ＋ ADM and BAI ＋ ADM significantly enhanced the susceptibility of HepG2 cells to CD3AK, with the killing rates increased from （30.00 ± 4.50） % in the control group to （97.23 ± 5.11 ） %, （93.12 ± 9.88 ） %, （60.45 ± 5.71 ） %, （43.87 ± 8.2）% and （47.25 ± 2.68）% , respectively（P 〈0.01 ）. Treatment wtih CD3AK also decreased the bcl-2 mRNA expression. Conclusion： ICA combined with BAI and ADM can inhibit the expression of APRIL and bcl-2 on HepG2 cells and subsequently inhibit the proliferation of HepG2 cells; meanwhile, it makes HepG2 cell more sensitive to CD3AK cells and reverses t