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顺铂在大鼠体内富集胃癌干细胞 被引量:4

Cisplatin therapy for in vivo enrichment of gastric cancer stem cells
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摘要 背景:肿瘤干细胞具有自我更新、耐药和转移成瘤能力,对肿瘤的发生、发展以及肿瘤的转移等具有重要的功能。目前,确认肿瘤干细胞的方法主要有2种,即体外瘤球培养实验以及小鼠体内成瘤实验,但临床上通过化疗在大鼠体内富集胃癌细胞尚缺乏研究报道。目的:观察顺铂在大鼠体内富集胃癌干细胞的情况,研究其表面标志蛋白的筛选方法。方法:建立BGC-823胃癌模型大鼠,采用随机对照方法将大鼠分为2组,实验组大鼠分别尾静脉注射质量浓度0.1,0.2,0.25,0.3 g/L的顺铂,而对照组则尾静脉注射生理盐水。经过3期化疗,最后得到富集胃癌干细胞组织;采用高通量蛋白芯片对肿瘤表面蛋白进行提取,并采用Western blot对芯片进行验证,比较顺铂在大鼠体内富集胃癌干细胞及其表面标志蛋白的筛选方法。结果与结论:顺铂剂量为0.3 g/L×200μL时治疗效果最佳,且化疗药物浓度越高,耐受力越差,不良反应发生率越高。移植瘤经苏木精-伊红染色验证均为癌组织;Western blot检测结果与蛋白芯片结果显示,HLA-DQ,PMP22和Claudin7蛋白在胃癌组织中表达增强,HLA-DR,CD14,CD16和CD56蛋白表达减弱。结果证实,顺铂化疗能够在体内富集胃癌干细胞,能够成功筛选出相应的表面标志物蛋白。 BACKGROUND: Tumor stem cells have self-renewal, drug resistance and metastasis tumorigenicity, which play an important role in occurrence, development and metastasis of tumors. Currently, there are two methods to identify tumor stem cells, namely, in vitro tumor sphere culture experiments and in vivo mouse tumorigenic experiments. However, there ia a lack of reports regarding clinically enriched gastric cancer cells by chemotherapy. OBJECTIVE: To investigate the enrichment of rat gastric cancer stem cells by cisplatin, and to explore the screening methods for their surface marker proteins. METHODS: BCG-823 gastric cancer model was established in rats, and then rat models were randomized into two groups: rats in experimental groups were subjected to intravenous injection of 0.1, 0.2, 0.25, 0.3 g/L cisplatin via the tail vein; those in control group were injected with normal saline via the tail vein. After three courses of chemotherapy, gastric stem cells-enriched tissues were collected. Tumor surface proteins were extracted using high-throughput protein microarray and identified by western blot assay. Effects of cisplatin on enrichment of rat gastric cancer stem cells and screening methods for surface marker proteins were compared.RESULTS AND CONCLUSION: Cisplatin at a dose of 0.3 g/L×200 μL exhibited the best therapeutic effects, and moreover, with the dose increasing, the tolerance became worse and the incidence of adverse reaction became higher. Transplantation tumors were verified by hematoxylin-eosin staining. Western blot test results were similar to the findings of protein microarray method, that is, HLA-DQ, PMP22 and Claudin7 protein expressions increased in gastric tissues, but HLA-DR, CD14, CD16 and CD56 protein expression decreased. These findings suggest that cisplatin can be used to enrich gastric cancer stem cells in rats, and to successfully screen the corresponding surface marker proteins.
出处 《中国组织工程研究》 CAS 北大核心 2015年第41期6611-6615,共5页 Chinese Journal of Tissue Engineering Research
基金 陕西省卫生厅科学研究基金(08H16)~~
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