蛴螬提取物体外对人MGC-803胃癌细胞株凋亡相关基因作用的研究

STUDY ON EFFECT OF GRUB EXTRACT ON APOPTOSIS RELATIVE-GENEOF MGC-803 HUMAN GASTRIC CANCER CELL LINE IN VITRO

目的 :探讨蛴螬提取物体外对MGC - 80 3胃癌细胞株的诱导凋亡作用的分子生物学机制。方法 :通过倒置显微镜、HE染色、吖啶橙 (AO) /溴化乙锭 (EB)荧光染色方法检测蛴螬提取物对MGC - 80 3胃癌细胞诱导凋亡和抑制作用的方式 ;应用免疫组织化学技术 (S -P法 )检测用药前后凋亡相关基因bcl- 2、bax表达的改变 ,探讨蛴螬提取物在体外对MGC - 80 3胃癌细胞株的抗增殖及诱导凋亡作用的可能机制。结果 :光镜结果表明 ,蛴螬 (终浓度为 4mg/ml)作用于MGC - 80 3胃癌细胞 2 4h后 ,可见胞核固缩、胞核碎裂、凋亡小体形成等凋亡形态学变化 ;经AO/EB荧光染色观察结果表明 ,当终浓度为 4mg/ml的蛴螬作用于MGC - 80 3细胞 2 4h后 ,其凋亡率和破膜率与自然凋亡率和破膜率相比均有显著性差异 ,其凋亡率为86 3% (P <0 0 0 1) ,破膜率为 4 1 9% (P <0 0 0 1)。免疫组化结果表明 ,用药前后凋亡相关基因bcl- 2、bax均有显著性改变 (P <0 0 0 1)。结论 :蛴螬提取物在体外对人MGC - 80 3胃癌细胞株有显著性抗增殖及诱导凋亡作用 ,其诱导肿瘤细胞凋亡的可能机制与凋亡相关基因bcl- 2、bas的表达改变有关。

Objective:To discuss the mechanism of inducing apoptosis of grub extract on MGC-803 gastric cancer cell line in vitro.Methods:Using invert microscope,HE stain and acridine orange(AO)/ethidium bromide(EB)fluorescence stain to examine the inhibiting proliferation and inducing apoptosis effct of grub extract on MGC-803 gastric cancer cells.The expression changes of relating-apoptosis gene proteins(bcl-2,bax) were detected by immunohistochemistry(S-P method) before and after treaed with the grub extract.Results:The grub extract significantly inhibited the proliferation of gastric cancer cells in vitro,morphologic changes of apoptosis with chromatin margination,karyopyknosis,karyorrhexis,apoptotic body were found by the light scope.It was showed by using acridine orange(AO)/ethidium bromide (EB) fluorescence staining that MGC-803 gastric cancer cells were not only necrosis but also apoptosis after treated by the grub extract,but the apoptosis was as the dominant way.The bcl-2 and bax of relating-apoptosis genes was significantly changed before and after treated with the grub extract( P <0 001).Conclusions:The grub extract can significantly inhibit the proliferation and induce apoptosis of MGC-803 gastric cancer cells in vitro and its possible mechanism of inducing apoptosis for tumor cells might be related with the expression changes of the bcl-2 and bax of relating-apoptosis genes.