双氢青蒿素对人卵巢癌细胞株HO-8910裸鼠移植瘤的作用及P53表达的影响

Effects of dihodryartemisinin on human ovarian cancer cell HO-8910 and P53 expression in nude mice implanted tumor

目的探讨双氢青蒿素(DHA)对人卵巢癌细胞株HO-8910裸鼠皮下移植瘤的作用及P53表达的影响。方法将人卵巢癌细胞株HO-8910注入裸鼠的腋下,建立人卵巢癌移植瘤的动物模型,随机分为空白组,低剂量DHA组、中剂量DHA组、高剂量DHA组、顺铂阳性对照组,观察干预后各组移植瘤抑瘤率;免疫组织化学法和蛋白印迹法检测肿瘤细胞内P53蛋白的表达;应用实时荧光定量聚合酶链反应(RT-PCR)法测定移植瘤中P53 mRNA的表达。结果①DHA对裸鼠移植瘤的生长有一定的抑制作用,并具有浓度依赖性。②病理学观察见细胞的体积较大,胞质疏松,核深染,可有不同程度的核固缩、核碎裂及核溶解,细胞中可见局灶性坏死。③免疫组织化学法结果证实随着DHA剂量的增加突变型P53蛋白表达减弱;蛋白印迹法检测到DHA组野生型P53蛋白表达上调。④RT-PCR法检测到DHA组野生型P53基因表达上调。结论 DHA对人卵巢癌裸鼠皮下移植瘤的生长有明显的抑制作用,且与剂量有一定的关系,其机制可能与上调野生型P53的表达导致细胞凋亡有关。

Objective To examine the effects of dihodryartemisinin （DHA） on human ovarian cancer cell HO- 8910 and P53 expression in nude mice subcutaneously implanted tumor. Methods The human ovarian cancer ceils H0-8910 were injected to nude mice＇s axillary to construct an animal model for H0-8910 implanted tumor, which was randomly assigned to blank group, low-dose DHA group, medium-dose DHA group, high-dose DHA group and cis- platin-positive contrast group for the determination of the implanted tumor growth inhibitory rate. Immunohistochemi- cal assay and Western blotting were employed to detect the level of P53 expression. And the reverse transcriptase polymerase chain reaction was applied to examine the P53-mRNA expression. Results DHA demonstrated inhibitory effects on tumor growth, in a concentration-dependent fashion. Pathological observation showed enlarged cells with loose cytoplasm and heavily stained nuclei, which were accompanied by the presence of pyknosis, nuclear dissolution, nuclear fragmentation, chromatin lumps and focal necrosis within the cells. Immunohistochemistry assay showed that the expression of mutation type P53 decreased towards increased dose of DHA. Western blotting confirmed that DHA resulted in increased expression of wide type P53, the mRNA of which was demonstrated to increase by reverse tran- scriptase polymerase chain reaction. Conclusion DHA confers inhibitory effects, which were associated with the dose administered, on subcutaneously transplanted tumor in nude mice, possibly related to the up-regulation of the wide type P53 expression leading to cell apoptosis.