青蒿琥酯对化疗耐药癌细胞裸鼠腮腺移植瘤c-Myc、ABCG2表达的影响

发布时间：2018-11-23 07:30

【摘要】：目的:探讨青蒿琥酯(Artesunate,ART)与顺铂(Cisplatin,DDP)单独给药或联合用药对裸鼠耐药细胞腮腺移植瘤的细胞增殖、耐药相关蛋白c-Myc及ABCG2及其m RNA表达的影响。方法:以人宫颈癌HELA细胞为亲本,经DDP低浓度、周期性体外诱导获得耐药子代癌细胞HELA-DDP,将耐药癌细胞接种于BALB/C-nu裸鼠一侧腮腺,建立耐药细胞腮腺移植瘤模型。接种7天后,触诊观察动物接种侧腮腺,如触及圆形或类圆形硬结,视为裸鼠腮腺移植瘤建模注射成功,而后应用随机表法将裸鼠分为生理盐水对照组、DDP 1.0 mg/kg组、DDP 2.0mg/kg组、ART 50.0 mg/kg、ART 100.0 mg/kg、ART 200.0 mg/kg组、联合用药组(ART 100 mg/kg+DDP 1.0 mg/kg组),分别给予腹腔注射给药,各组隔天给药一次。给药30天后处死动物,获取瘤体标本,所获其余实验数据另文撰写。免疫组化法检测各组裸鼠腮腺移植瘤组织中c-Myc与ABCG2蛋白的表达水平;Western Blot法、实时荧光定量PCR法分别检测各组裸小鼠腮腺移植瘤组织中c-Myc与ABCG2基因的m RNA及蛋白表达水平。结果:1.实时荧光定量PCR结果显示:ART从50 mg/kg剂量组至200 mg/kg剂量组均可抑制耐药癌细胞裸鼠腮腺移植瘤中c-Myc与ABCG2 m RNA的表达,与生理盐水组相比差异均具有显著性(P0.01),与顺铂组相比差异均具有显著性(P0.05)。2.免疫组化结果显示:ART从50 mg/kg剂量组至200 mg/kg剂量组均可降低耐药癌细胞裸鼠腮腺移植瘤组织中c-Myc与ABCG2蛋白表达的平均光密度值,与生理盐水组相比差异具有显著性(P0.05),与顺铂组相比差异具有显著性(P0.05)。3.Western Blot结果显示:ART从50 mg/kg剂量组至200 mg/kg剂量组均可降低耐药癌细胞裸鼠腮腺移植瘤组织中c-Myc与ABCG2蛋白表达水平,与生理盐水组相比差异均具有显著性(P0.05)。结论:1.ART具有抑制耐药癌细胞裸鼠移植瘤细胞增殖及逆转其耐药的作用。其可能与ART下调裸鼠移植瘤组织内c-Myc与ABCG2 m RNA和蛋白表达有关。2.ART和DDP联合用药时,ART既起到抑制肿瘤细胞增殖的作用,还起到逆转化疗耐药性的作用。
[Abstract]:Aim: to investigate the effects of artesunate (Artesunate,ART) and cisplatin (Cisplatin,DDP) alone or in combination on the expression of c-Myc, ABCG2 and m RNA in parotid gland xenografts in nude mice. Methods: human cervical cancer HELA cells were used as parents. Drug resistant progeny cancer cells (HELA-DDP,) were induced by low concentration of DDP in vitro. The drug-resistant cancer cells were inoculated into the parotid gland of BALB/C-nu nude mice, and the model of drug resistant cell parotid gland transplantation tumor was established. After 7 days of inoculation, palpation and observation of the parotid glands of the inoculated side of the animals, if they touched round or round knots, were regarded as successful modeling and injection of the transplanted tumors of the parotid gland in nude mice. Then the nude mice were divided into normal saline control group and DDP 1.0 mg/kg group by random table method. DDP 2.0mg/kg group, ART 50.0 mg/kg,ART 100.0 mg/kg,ART 200.0 mg/kg group, combined treatment group (ART 100 mg/kg DDP 1.0 mg/kg group), were given intraperitoneal injection, once every other day. After 30 days of administration, animals were killed and tumor specimens were obtained. The remaining experimental data were written separately. Immunohistochemical method was used to detect the expression of c-Myc and ABCG2 protein in the tissues of parotid gland xenografts in each group. The expression levels of m RNA and protein of c-Myc and ABCG2 genes in parotid gland xenografts of nude mice were detected by Western Blot and real-time fluorescence quantitative PCR. Results: 1. The results of real-time fluorescence quantitative PCR showed that ART could inhibit the expression of c-Myc and ABCG2 m RNA in xenografts of parotid gland in nude mice from 50 mg/kg to 200 mg/kg. There were significant differences compared with normal saline group (P0.01) and cisplatin group (P0.05). Immunohistochemical results showed that the average optical density of the expression of c-Myc and ABCG2 protein in parotid gland transplanted tumor tissues of nude mice was decreased by ART from 50 mg/kg to 200 mg/kg. Compared with saline group, the difference was significant (P0.05). Compared with cisplatin group, there was significant difference (P0.05). 3.Western Blot results showed that the expression of c-Myc and ABCG2 protein in parotid gland xenografts of nude mice with drug-resistant cancer cells could be decreased by ART from 50 mg/kg to 200 mg/kg. Compared with saline group, there were significant differences (P0.05). Conclusion: 1.ART can inhibit the proliferation and reverse the drug resistance of xenografts of drug resistant cancer cells in nude mice. It may be related to the down-regulation of c-Myc and ABCG2 m RNA and protein expression in xenografts of nude mice by ART. When combined with 2.ART and DDP, ART can not only inhibit the proliferation of tumor cells, but also reverse the chemotherapeutic resistance.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.8

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