大蒜素对皮下移植肝癌细胞生长抑制和诱导凋亡的实验研究

Effects of allicin on the proliferation and apoptosis of transplanted hepatocellular carcinoma

目的 评价大蒜素在实验裸小鼠体内抗肝癌的有效性,为临床应用大蒜素对肝癌进行介入治疗提供理论依据。 方法 建立人肝癌细胞BEL 7402裸小鼠皮下移植瘤模型,模拟介入治疗用药方法,对6组移植瘤模型分别注射生理盐水吐温溶液,1、2、5 mg/ml大蒜素,0 .2 mg/ml阿霉素,1 mg/ml大蒜素+0 .2 mg/ml阿霉素,每次注射 0. 1 ml,隔日 1 次,连续 7 次,测量肿瘤体积变化,计算各组抑瘤率,对移植瘤组织作HE染色观察,电镜观察形态改变, TUNEL法检测组织凋亡率,RT PCR技术分析处理前后BEL 7402细胞FasL mRNA表达。 结果 移植瘤生长符合指数曲线规律,各组抑瘤率分别为: 62 .8%、72 .8%、94 .5%、76. 0%和 87 .0%;各组凋亡指数分别为(2. 2±1 .1)%、(17 .2±2. 8)%、(19 .4±2 .1)%、(28. 3±3.0)%、(22. 4±2 .5)%和(37 .6±3 .9)%;大蒜素或阿霉素组FasL mRNA表达均增加,大蒜素+阿霉素组表达上调尤其明显;大蒜素或(和)阿霉素治疗后,移植瘤组织内坏死细胞增加,出现典型凋亡形态细胞。 结论 大蒜素可以有效抑制人肝癌细胞裸小鼠移植瘤生长,且呈剂量效应关系;大蒜素可通过上调FasL mRNA的表达诱导人肝癌细胞凋亡。

Objective To evaluate the therapeutic effect of allicin on heapatocellular carcinoma in vivo and to furnish theoretical and methodological basis for the clinical application of allicin in interventional therapy of heaptocellular carcinoma. Methods The transplanted tumor model was established by injecting human hepatocellular carcinoma BEL -7402 cell into subcutaneous tissue of nude mice. The growth curve of these transplanted tumor cells was measured. Thirty nude mice were divided into 6 groups at random: 0.1 ml of normal saline tween solution,1 mg/ml allicin,2 mg/ml allicin,5 mg/ml allicin,1 mg/ml allicin plus 0.2 mg/ml adriamycin directly injected into tumor body respectively by interventional methods, for seven times at an interval of two days. The changes of tumor volume,the tumor inhibition rate, the morphologic alterations under light microscope by HE staining and electron microscope, and the apoptosis rate with TUNEL assay were observed. The expression of apoptosis-associated FasL mRNA before and after above treatments by using of RT-PCR assay was analyzed. Results The growth curve of transplanted BEL-7402 cells was exponential.The inhibition rates were 62.8%,72.8%,94.5%,76.0% and 87.0% respectively in the above groups.The apoptosis indices were (2.2±1.1)%,(17.2±2.8)%,(19.4±2.1)%,(28.3±3.0)%,(22.4±2.5)% and (37.6±3.9)%,respectively.The expression of FasL mRNA was remarkably up-regulated in allicin group and adriamycin group, especially in allicin plus adriamycin group.The increased necrotic cells and typical apoptosis cells in the transplanted tumor tissue under electron microscope were observed in the allicin or (and) adriamycin group. Conclusions Allicin can inhibit the growth of transplanted hepatocellular carcinoma cells effectively in vivo by interventional method, exhibiting a dose-dependent relationship. Allicin may induce apoptosis of transplanted human hepatocellular carcinoma cells in vivo and up-regulate the expression of FasL mRNA.