藤梨根提取物抗肺癌作用的体内实验研究

Inhibitory effect of Radix Actinidiae extractive on transplantation tumor of lung cancer cell line A549 in mude mice and its mechanism

目的研究藤梨根乙酸乙酯提取物对肺癌A549细胞裸鼠移植瘤的生长抑制及凋亡诱导作用,探讨其可能机制。方法用肺癌A549细胞建立肺癌裸鼠移植瘤模型,随机分为对照组、治疗组(高剂量组、低剂量组)。根据瘤体积变化绘制移植瘤生长曲线,根据终末瘤质量比较计算抑制率,移植瘤细胞凋亡指数检测采用TUNEL法,移植瘤细胞Bcl-2、Bax蛋白、Ki-67抗原表达检测采用免疫组化SP法,RT-PCR检测移植瘤Bcl-2、Bax mRNA表达。结果各治疗组经藤梨根乙酸乙酯提取物治疗后移植瘤生长缓慢,治疗结束时治疗组移植瘤质量及瘤体积明显低于对照组(P均<0.01),高、低剂量组的瘤质量抑制率分别为69.00%和45.09%。治疗组移植瘤细胞透射电镜下均可见不同阶段典型的凋亡细胞形态学变化。各治疗组移植瘤组织凋亡指数明显高于对照组,而增殖指数明显减低,Bcl-2 mR-NA及蛋白表达较对照组亦明显减少,Bax蛋白及mRNA表达较对照组明显增加,高剂量组尤其明显(P均<0.01)。结论藤梨根乙酸乙酯提取物对肺癌A549细胞裸鼠移植瘤生长有抑制作用,其机制与抑制移植瘤细胞的增殖活性和诱导癌细胞凋亡有关,而降低Bcl-2基因表达、上调Bax基因表达可能是其诱导细胞凋亡机制之一。

Objective To study the inhibitory effect and induced apoptosis function of acetidin extractive of radix Actinidiae on transplantation tumor of lung cancer A549 cell in mude mice,and to explore its mechanism of action.Methods Establishing the modle of transplantation tumor bearing lung cancer A549 cells in mude mice to detect the effect of acetidin extractive of radix Actinidiae in vivo.Control group and treatment groups（high-dose group and low-dose group）were set for comparison.The transplantation tumor growth curve was drawn according to transplantation tumor volume,and inhibition ratio was calculated according to the last tumor weight.Terminal deoxynucleotidyl transferase mediated UTP Nick End Labeling（TUNEL） was used to examine the apoptosis index of the transplantation tumor.Immunohistochemistry（SP） was used to examine the expressions of Bcl-2,Bax protein and Ki-67 antigen in the transplantation tumor.RT-PCR was used to examine the expressions of Bcl-2,Bax mRNA.Results The growth of transplantation tumor in treatment groups became slowly after taking drug.The last tumor volume and tumor weight of treatment groups were smaller than that of control group.The tumor weight inhibiting rates of high-dose group and low-dose group were 69% and 45.09% individually.Transmission electron microscope also detect transplantation tumor cells to undergo apoptosis with typically apoptotic characteristics.The apoptosis index of the transplantation tumor of treatment groups was higher than that of control group.The expressions of Bcl-2 protein,Ki-67 antigen in treatment groups were lower than those of control group.The expressions of Bax in treatment groups were higher than those of control group especialy the high-hose group（P0.01）.Conclusions The acetidin extractive of radix Actinidiae has the inhibitory effect on the growth of transplantation tumor of lung cancer A549 cell in mude mice.To inhibit proliferation activity of transplantation tumor,to degrade the expression of Bcl-2,to up-regulation the expression of Bax,and to induce apoptosis of cancer cell may be its mechanism of action.